Staphylococcus spp., Streptococcus canis,and Arcanobacterium phocae of healthy Canadian farmed mink and mink with pododermatitis

Pododermatitis is a disease of concern for mink breeders in Canada and worldwide, as it causes discomfort and lowers the breeding rates on farms affected by the disease. Unfortunately, the etiology and pathogenesis of pododermatitis are still unknown. In this study, we compared Staphylococcus spp. and Streptococcus canis isolates from healthy mink with isolates from animals with pododermatitis on 2 farms in Ontario. Almost all hemolytic Staphylococcus spp. isolated were shown to be Staphylococcus delphini Group A by 16S ribosomal ribonucleic acid (rRNA) sequence analysis and polymerase chain reaction (PCR). Pulsed-field gel electrophoresis (PFGE) did not reveal any S. delphini or S. canis clonal lineages specifically associated with pododermatitis, which suggests that these bacteria do not act as primary pathogens, but does not dismiss their potential roles as opportunistic pathogens. While S. delphini and S. canis were the most prevalent bacterial pathogens in mink pododermatitis, they were also present in samples from healthy mink. Arcanobacterium phocae is occasionally isolated from pododermatitis cases, but is difficult to recover with conventional culture methods due to its slow growth. A quantitative real-time PCR was developed for the detection of A. phocae and was tested on 138 samples of footpad tissues from 14 farms. The bacterium was detected only in pododermatitis-endemic farms in Canada and was at higher concentrations in tissues from infected footpads than in healthy tissues. This finding suggests that A. phocae is involved in the pathogenesis of pododermatitis.     

Lack of a type-2 glycosyltransferase in the fish pathogen Flavobacterium psychrophilum determines pleiotropic changes and loss of virulence

Flavobacterium psychrophilum is an important fish pathogen, responsible for Cold Water Disease, with a significant economic impact on salmonid farms worldwide. In spite of this, little is known about the bacterial physiology and pathogenesis mechanisms, maybe because it is difficult to manipulate, being considered a fastidious microorganism. Mutants obtained using a Tn4351 transposon were screened in order to identify those with alteration in colony morphology, colony spreading and extracellular proteolytic activity, amongst other phenotypes. A F. psychrophilum mutant lacking gliding motility showed interruption of the FP1638 locus that encodes a putative type-2 glycosyltransferase (from here on referred to as fpgA gene, Flavobacterium psychrophilum glycosyltransferase). Additionally, the mutant also showed a decrease in the extracellular proteolytic activity as a consequence of down regulation in the fpgA mutant background of the fpp2-fpp1 operon promoter, responsible for the major extracellular proteolytic activity of the bacterium. The protein glycosylation profile of the parental strain showed the presence of a 22 kDa glycosylated protein which is lost in the mutant. Complementation with the fpgA gene led to the recovery of the wild-type phenotype. LD₅₀ experiments in the rainbow trout infection model show that the mutant was highly attenuated. The pleiotropic phenotype of the mutant demonstrated the importance of this glycosyltranferase in the physiology and virulence of the bacterium. Moreover, the fpgA mutant strain could be considered a good candidate for the design of an attenuated vaccine.     

A preliminary trial of the sedation induced by intranasal administration of midazolam alone or in combination with dexmedetomidine and reversal by atipamezole for a short‐term immobilization in pigeons

ObjectiveTo assess the sedative and immobilization effect of intranasal administration (INS) of midazolam (MID) without or with INS dexmedetomidine (DXM), and some physiological changes induced by the drugs. The ability of INS atipamezole to reverse the DXM component was also assessed.Study designProspective ‘blinded’ experimental study.AnimalsIn total, 15 pigeons.MethodsPigeons were sedated by INS MID alone at a dose of 5 mg kg⁻¹ (group MID, n = 6) or in combination with INS DXM at a dose 80 μg kg⁻¹ (group MID-DXM, n = 6). Measurements were made of heart rate (HR), respiratory rate (f_R) and cloacal temperature (CT). The degree of sedation was assessed at 15 minutes prior to, immediately after, and at intervals until 100 minutes after drug administrations. Following MID-DXM, INS atipamezole (250 μg kg⁻¹) was administered and the same indices measured 5 and 10 minutes later.ResultsMID had no effect on HR and f_R, and although CT decreased, it remained within physiological range. MID-DXM caused significant falls in HR, f_R and CT that persisted until the end of sedation. Atipamezole antagonized sedation and cardiorespiratory side effects of MID-DXM within 10 minutes of application. In addition, for MID compared to MID-DXM, the lowest sedation scores [10 (7–14) and 10.5 (5–14) versus 2 (1–4) and 2 (1–5)] were achieved in the 10th and 20th minute versus the 20th and 30th minute of the sedation, respectively.Conclusions and clinical relevanceMID alone, given INS had minimal side effects on vital functions but caused inadequate immobilization of pigeons for restraint in dorsal recumbency. MID-DXM caused an effective degree of immobilization from 20 to 30 minutes after administration, at which time birds tolerated postural changes without resistance. Atipamezole antagonized both side effects and sedation, but complete recovery had not occurred within 10 minutes after its application.     

Detection of castor contamination by real-time polymerase chain reaction

Due to the potential for intentional contamination of food with crude preparations containing ricin, a real-time PCR method was developed for the detection of castor plant material in ground beef. One primer pair was identified and confirmed to be castor-specific and efficient for amplification of ricin in DNA extracts from castor or beef matrices. Of three different DNA extraction protocols compared, the hexadecyltrimethylammonium bromide (CTAB) method yielded the highest quality of DNA for QPCR assay. The detection limit for castor contamination in ground beef samples was <0.001% (<10 microg of castor acetone powder per gram of beef, corresponding to 0.5 microg of ricin), indicating excellent sensitivity for the assay, well below the threshold for oral toxicity.     

ATR-Fourier transform mid-infrared spectroscopy for determination of trans fatty acids in ground cereal products without oil extraction

Fourier transform mid-infrared (FT-IR) spectroscopy was investigated as a method of analysis for trans fatty acid content of cereal products without the need for prior oil extraction. Spectra were obtained, with an FT-IR spectrometer equipped with an attenuated total reflectance (ATR) device, of ground samples pressed onto the diamond ATR surface, and trans fatty acids were measured by a modification of AOAC Method 996.01. Partial least-squares (PLS) models were developed for the prediction of trans fatty acids in ground samples using several wavenumber selections on the basis of bands related to lipids. The models (n = 79) predicted trans fatty acids in ground samples with standard error of cross-validation (SECV) of 1.10-1.25 (range 0-12.4) % and R2 of 0.85-0.88 and in validation samples (n = 26) with standard error of performance (SEP) of 0.96-1.12 (range 0-12.2) % and r2 of 0.89-0.92, indicating sufficient accuracy for screening. Sample trans fatty acid % was predicted as accurately with the fingerprint region (1500-900 cm(-1)) as with the entire range (4000-650 cm(-1)) indicating, in concert with the regression coefficients, the importance of the isolated trans double bonds at 966 cm(-1) in development of the model. Data is also presented on prediction of trans fatty acids using the spectra of residual oil films on the ATR surface after removing the solid portion of the sample.     

Formation of hydrogel particles by thermal treatment of beta-lactoglobulin-chitosan complexes

Molecular complexes based on proteins and ionic polysaccharides have considerable potential for encapsulation of functional food components, but their widespread utilization is limited because their structure is highly sensitive to pH and ionic strength. We have investigated the possibility of creating stable hydrogel particles by thermal treatment of protein (beta-lactoglobulin) and cationic polysaccharide (chitosan) mixtures. Mixed solutions of beta-lactoglobulin (0.5 wt %) and chitosan (0.1 wt %) were prepared at various pH's (3-8) and were heated (80 degrees C for 20 min). Prior to heating, the biopolymer mixtures formed molecular complexes at pH values where there was an electrostatic attraction between the protein and the polysaccharide: soluble complexes at pH 4.5; complex coacervates at pH 5.0 and 5.5; precipitates at pH>5.5. After heating, relatively small (d approximately 140 nm) and cationic (zeta>+20 mV) hydrogel particles were formed at pH 4.5, but much larger aggregates were formed at pH 5.0 and higher (d>1000 nm). The thermally treated hydrogel particles formed at pH 4.5 maintained their initial particle size when the pH was subsequently adjusted within the range pH 3-5, but they aggregated when the pH was adjusted to >pH 5 because of a reduction in the magnitude of their electrical charge. This study suggests that hydrogel particles can be formed by heating mixed protein-polysaccharide systems under controlled conditions. These hydrogel particles may be useful for encapsulation of functional food components.     

A systematic and quantitative approach to improve water use efficiency in agriculture

As the competition for the finite water resources on earth increases due to growth in population and affluence, agriculture is faced with intensifying pressure to improve the efficiency of water used for food production. The causes for the relatively low water use efficiency in agriculture are numerous and complex, including environmental, biological, engineering, management, social, and economic facets. The complexity of the problem, with its myriads of local variations, requires a comprehensive conceptual framework of the underlying physical and biological processes as the basis to analyze the existing situation and quantify the efficiencies, and to plan and execute improvements. This paper proposes such a framework, based on the simple fact that the overall efficiency of any process consisting of a chain of sequential step is the product of the efficiency (i.e., output/input ratio) of its individual component steps. In most cases of water use, a number of process chains, both branching and merging, are involved. Means to integrate the diverging and converging chains are developed and presented as equations. Upscaling from fields to regions and beyond are discussed. This chain of efficiencies approach is general and can be applied to any process composed of chains of sequential steps. Here the framework is used to analyze the systems of irrigated and dryland crop production, and animal production on rangeland. Range of plausible efficiencies of each step is presented as tables, with values separately for the poor and for the good situation of circumstances, management and technology. Causes of the differences in efficiency of each step, going from water delivery to soil water extraction, transpiration, photosynthesis, and conversion to crop biomass and yield, and to animal product are briefly discussed. Sample calculations are made to demonstrate how modest differences in the efficiencies of the component steps are manifested as large to huge differences in the overall efficiency. Based on an equation quantifying the impact of changes in efficiency of component steps on the overall efficiency, it is concluded that generally, it is more effective to made modest improvements in several or more steps than to concentrate efforts to improve one or two steps. Hence, improvement efforts should be systematic and not overly concentrated on one or two components. The potential use of the same equation as the point of departure to optimize the allocation of economic resource among the component steps to maximize the improvement in the overall water use efficiency is elaborated on. The chain of efficiencies framework provides the means to examine the current levels of efficiency along the pathways of agricultural water use, to analyze where inefficiencies lie by comparing with the range of known efficiency values in the tables presented, to assess the potential improvements that may be achieved in various parts and their impact on the overall efficiency, and to aid in the optimal allocation of resources for improvements.

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