Development and evaluation of a noninvasive marker cluster technique to assess three-dimensional kinematics of the distal portion of the forelimb in horses

OBJECTIVE: To develop and evaluate a marker cluster set for measuring sagittal and extrasagittal movement of joints in the distal portion of the forelimb in ponies. ANIMALS: 4 ponies. PROCEDURES: 5 infrared cameras were positioned on a concrete walkway in a frontal-sagittal arc and calibrated. Four segments were defined: hoof, middle phalanx, proximal phalanx, and metacarpus. Rigid clusters with 4 retroreflective markers were placed on each segment. A static trial was recorded with additional anatomic markers on the medial and lateral joint lines. Those anatomic markers were removed, and kinematic data were recorded at 240 Hz during walking. An ensemble mean was computed from the 4 ponies from 5 replicates of the walks. Joint kinematic variables were calculated by use of the calibrated anatomical system technique. The design and error dispersion of each marker were evaluated. RESULTS: Marker clusters were quasiplanar, but variation in orientation error was reduced because the mean radii were > 10 times the largest error dispersion values. Measurements of sagittal rotations of the distal interphalangeal, proximal interphalangeal, and metacarpophalangeal joints were similar to measurements obtained with bone-fixed triads, but larger discrepancies between the 2 methods were found for extrasagittal rotations. CONCLUSIONS AND CLINICAL RELEVANCE: Development of noninvasive methods for quantifying data pertaining to 3-dimensional motion in horses is important for advancement of clinical analysis. The technique used in the study enabled identification of flexion-extension motions with an acceptable degree of accuracy. Appropriate correction algorithms and improvements to the technique may enable future quantification of extrasagittal motions.     

Partitioning and inhibition of lipid oxidation in mechanically separated turkey by components of cranberry press cake

Extracts from cranberry press cakes were prepared either using ethanol or an ethyl acetate-acetone mixture. The press cake extracts were compared with extracts from cranberry juice powder (CJP), prepared using chloroform:methanol (1:1), for their ability to inhibit lipid oxidation in mechanically separated turkey (MST). Because of the susceptibility of muscle membrane lipids to oxidation, the ability of quercetin in the extracts to partition between the aqueous and the membrane phases was studied. Membrane suspensions were prepared from MST. Partitioning of quercetin was quantified using high-performance liquid chromatography. Oxidation was studied by measuring thiobarbituric acid reactive substances and lipid peroxides. The effectiveness of the extracts to inhibit lipid oxidation was CJP extract > ethyl acetate extract of press cake > or = ethanol extract of press cake. The amount of quercetin in the extracts and the amount of quercetin that partitioned into the membranes followed the same order. However, the total phenolic content of the extracts did not follow the same order as that of inhibitory power. The phenolic content of the extracts decreased, ethyl acetate extract > ethanol extract of press cake > or = chloroform extract of CJP. Irrespective of the extraction method, around 78% quercetin from the extracts partitioned into the membranes. It could be concluded that increasing the amount of quercetin in the press cake extracts increases the ability of the extracts to inhibit lipid oxidation in MST. Hence, a proper choice of solvents and extraction method, which would increase the amount of quercetin in the press cake extracts, might increase the antioxidant potential of the extracts and hence their economic value.     

Identification of lactoferrin and glutamate receptor‐interacting protein 1 in bovine cervical mucus: A putative marker for oestrous detection

Accurate detection of oestrus is important for artificial insemination. The aim of this study was to identify oestrous‐specific bovine cervical mucus proteins that could be used to determine the optimal time for artificial insemination. Non‐oestrous and controlled internal drug release (CIDR)‐induced oestrous‐stage mucus proteins were purified and subjected to surface‐enhanced laser desorption/ionization time‐of‐flight mass spectrometry, sodium dodecyl sulphate polyacrylamide gel electrophoresis and MALDI‐TOF/TOF. Among differentially expressed proteins, lactoferrin (LF) and glutamate receptor‐interacting protein 1 (GRIP1) showed a twofold increase during the CIDR‐induced oestrous stage compared to the levels in non‐oestrous stage in bovine cervical mucus. The RT‐PCR, Western blotting and immunohistochemistry results showed that LF and GRIP1 expression was significantly increased during the oestrous stage in the uterus. This study demonstrated that bovine LF and GRIP1 exist during the oestrous stage, but not during the non‐oestrous stage, suggesting that cervical mucus LF and GRIP1 are useful oestrous detection markers in cattle.     

Assessing genetic diversity of wild southeastern North American <Emphasis Type="Italic">Vaccinium</Emphasis> species using microsatellite markers

Wild species representatives from Northwestern, Central and Southern Florida, and neighboring U.S. states were collected in multiple United States Department of Agriculture (USDA) exploration expeditions and are being preserved at the USDA, Agricultural Research Service, National Clonal Germplasm Repository in Corvallis, Oregon. Germplasm from these southeastern regions of North America is particularly vulnerable to loss in the wild due to encroachment of human development in key habitats and biotic and abiotic stresses from climate change. Fourteen simple sequence repeats (SSRs), previously developed from the highbush blueberry (Vaccinium corymbosum) cultivar ‘Bluecrop’, were used to estimate genetic diversity and genetic differentiation of 67 diploid individuals from three species, including 19 V. elliottii, 12 V. fuscatum, and 35 V. darrowii accessions collected throughout the species’ ranges. Results from our analyses indicated that the samples from each species could be reliably resolved using genetic distance measures with ordination and neighbor joining approaches. In addition, we estimated admixture among these species by using Bayesian assignment tests, and were able to identify a mis-labeled accession of V. darrowii ‘Johnblue’, two mis-classified accessions (CVAC 735.001 and CVAC 1223.001), and four accessions of previously undescribed hybrid origin (CVAC 734.001, CVAC 1721.001, CVAC 1741.001, and Florida 4B CVAC 1790). Allele composition at the 14 SSRs confirmed that Florida 4B CVAC 1790, the donor of low chilling for the southern highbush blueberry, was the critical parent of US 74. Genetic diversity assessment and identification of these wild accessions are crucial for optimal germplasm management and expand opportunities to utilize natural variation in breeding programs.     

Yield response of two potato culivars to supplemental irrigation and N fertilization in New Brunswick

Nitrogen and water are important factors influencing potato production, and crop response to these two factors may vary with cultivars. The yield response of two potato cultivars (Russet Burbank and Shepody) to six rates of N fertilization (0-250 kg N ha^-1) with and without supplemental irrigation was studied at four onfarm sites in each of three years, 1995 to 1997, in the upper St-John River Valley of New Brunswick, Canada. On average, irrigation increased total yield from 31.9t ha^-1 without irrigation to 38.41 ha^-1 with irrigation and marketable yield from 25.61 ha^-1 without irrigation to 30.71 ha^-1 with irrigation. Potato yields were increased by irrigation at nine out of the 12 sites, and the irrigation response was similar for both cultivars. Nitrogen fertilization significantly increased both total and marketable yields at all sites except one. The yield response to N fertilization was greater with irrigation. The N fertilization rate (Nmax) required to reach maximum total and marketable yield, however, was similar with and without irrigation. A large variation in Nmax was observed among sites. With irrigation Nmax varied between 158 and 233 kgN ha^-1 for total yield, and between 151 and 250 kg N ha^-1 for marketable yield. There was no interaction between N fertilization and potato cultivar for both total and marketable yields. The two cultivars had similar total yields (35 t ha^-1). Shepody, however, had a greater marketable yield (28.9 t ha^-1) than Russet Burbank (27.4 t ha^-1). Our results indicate that the response to two of the most significant factors of potato production, irrigation and N fertilization, varies greatly with sites and climatic conditions, and that field specific recommendations are required for the optimum management of N and irrigation.     

Immunohistochemical and PCR studies of wild fish for Tetracapsula bryosalmonae (PKX), the causative organism of proliferative kidney disease

Tetracapsula bryosalmonae, previously referred to as PKX, causes proliferative kidney disease (PKD) in salmonids and is an economically important myxozoan pathogen in salmonid culture. A variety of molecular and immunological tools have been developed to detect the parasite. To determine the specificity of four monoclonal antibodies (MAbs) raised against T. bryosalmonae, archive material of fish infected with various myxosporean species was obtained and immunostained. Wild fish were also collected from enzootic waters and examined for T. bryosalmonae infection using immunohistochemistry and the polymerase chain reaction (PCR). Three of the MAb probes appear to be specific for T. bryosalmonae while only two of the five sets of primers tested appeared to specifically amplify T. bryosalmonae DNA. The results of the immunostaining and the PCR demonstrate that T. bryosalmonae occurs in the tubules of grayling Thymallus thymallus L., brown trout, Salmo trutta L. and Atlantic salmon, Salmo salar L. outside of the PKD season (June‐September) in the UK. This confirms the results of previous studies that these species are the preferred fish hosts for the parasite in the UK.     

Atrazine intoxication in cattle,confirmed by gas chromatography–mass spectrometry

Ten of 40 cows died within 48 h of gaining access to a barn in which various chemicals were stored. Some of the surviving cows exhibited drooling, muscle tremors, and agitation. Postmortem examinations of 2 cows were performed in the field, and revealed nonspecific, moderate-to-severe pulmonary congestion. Liver and rumen contents, each from a different cow, were analyzed using a qualitative, multi-residue GC-MS method validated for the detection of pesticides and other chemical analytes. Using this method, extracts from the liver and rumen content samples were compared to atrazine (neat standard) and matrix-matched, control samples fortified with atrazine. GC-MS analysis detected atrazine at 215 m/z (NIST match >97%) with a retention time of ~13 min in liver and rumen content samples from our case. Detection of atrazine in the samples from the cows in this herd, combined with the clinical history, indicate that atrazine toxicity was the likely cause of clinical signs and death observed in this herd.