Assessment of PCR-based simple sequence repeats to fingerprint North American potato cultivars

Seven primer pairs flanking di- and tri-nucleotide repeat sequences, identified from previously sequenced regions of the potato genome, were examined for their potential use in DNA-fingerprinting of thirty-nineSolanum tuberosum subsp. tuberosum cultivars (released between 1861 and 1988) and one diploidS. phureja breeding line. Of the simple sequence repeats (SSRs), the primers for six SSRs amplified DNA sequences within the potato genome between cultivars for a total of 14 bands. The polymerase chain reaction (PCR)based amplification products generated from each primer pair consisted of 1 to 2 bands per cultivar but band variation among cultivars demonstrated up to 4 bands per SSR. A similarity matrix generated from five SSRs was able to distinguish 24 of the 40 cultivars. However, when the potato cultivars were grouped by tuber type (round white-skinned, long white-skinned, russetskinned, red-skinned, and yellow flesh) only five pairs of cultivars remained indistinguishable: Atlantic/Katahdin, Belchip/Wauseon, Red LaSoda/Bliss Triumph, Red Pontiac/Norland, and Burbank/Spunta. Although SSRs did not generate unique fingerprints for all of the North American genotypes examined, the potential to discriminate most cultivars should increase as additional SSRs are identified in potato.     

A Comparative Study of Non-contact Infrared and Digital Rectal Thermometer Measurements of Body Temperature in the Horse

Equine body temperature taken with a non-contact infrared thermometer (NCIT) does not correlate well with the equine body temperature taken with a digital rectal thermometer (DRT), when it comes to absolute temperatures. There is significant variation in body temperature readings depending on the site at which the temperature was taken. NCIT may still may have, in that it does seem to be able to reliably detect when temperatures are more than that which is clinically normal, and does not seem to give falselyelevated readings in clinically normal horses. In horses that resent having their temperatures taken rectally, the NCIT seems to offer the equine clinician a quick and easy way to at least determine whether the horse's temperature is at or more than normal, even if it does not agree with DRT results.     

Dimension reduction and variable selection for genomic selection: application to predicting milk yield in Holsteins

Genome‐assisted prediction of genetic merit of individuals for a quantitative trait requires building statistical models that can handle data sets consisting of a massive number of markers and many fewer observations. Numerous regression models have been proposed in which marker effects are treated as random variables. Alternatively, multivariate dimension reduction techniques [such as principal component regression (PCR) and partial least‐squares regression (PLS)] model a small number of latent components which are linear combinations of original variables, thereby reducing dimensionality. Further, marker selection has drawn increasing attention in genomic selection. This study evaluated two dimension reduction methods, namely, supervised PCR and sparse PLS, for predicting genomic breeding values (BV) of dairy bulls for milk yield using single‐nucleotide polymorphisms (SNPs). These two methods perform variable selection in addition to reducing dimensionality. Supervised PCR preselects SNPs based on the strength of association of each SNP with the phenotype. Sparse PLS promotes sparsity by imposing some penalty on the coefficients of linear combinations of original SNP variables. Two types of supervised PCR (I and II) were examined. Method I was based on single‐SNP analyses, whereas method II was based on multiple‐SNP analyses. Supervised PCR II was clearly better than supervised PCR I in predictive ability when evaluated on SNP subsets of various sizes, and sparse PLS was in between. Supervised PCR II and sparse PLS attained similar predictive correlations when the size of the SNP subset was below 1000. Supervised PCR II with 300 and 500 SNPs achieved correlations of 0.54 and 0.59, respectively, corresponding to 80 and 87% of the correlation (0.68) obtained with all 32 518 SNPs in a PCR model. The predictive correlation of supervised PCR II reached a plateau of 0.68 when the number of SNPs increased to 3500. Our results demonstrate the potential of combining dimension reduction and variable selection for accurate and cost‐effective prediction of genomic BV.     

Salmonella Typhimurium invasion of HEp-2 epithelial cells in vitro is increased by N-acylhomoserine lactone quorum sensing signals

Background

In Gram-negative bacteria, the most commonly studied quorum sensing signals are the N-acylhomoserine lactones (AHLs). In Salmonella, AHLs are recognized by SdiA, which is believed to be a sensor of AHLs produced by other bacteria, since Salmonella does not produce AHLs itself. It has been speculated that AHLs produced by the gastrointestinal flora may influence the regulation of virulence traits in Salmonella. The aim of the present work was to study the effect of AHLs on epithelial cell invasion by Salmonella in vitro.

Methods

Invasion by Salmonella enterica subspecies enterica serovar Typhimurium (S. Typhimurium) strain and its isogenc sdiA mutant was studied using a conventional gentamycin invasion assay with HEp-2 cells at 37°C. Gene expression was studied using a semi-quantitative PCR.

Results

The S. Typhimurium strain, but not its isogenic sdiA mutant, displayed increased in vitro invasion after addition of both N-hexanoyl-DL-homoserine lactone (C6-AHL) and N-octanoyl-DL-homoserine lactone (C8-AHL). Increased expression of two of the genes in the SdiA regulon (rck and srgE) was observed in the wild type strain, but not in the sdiA mutant.

Conclusions

The results from the present study show that S. Typhimurium can respond to two different AHL quorum sensing signals (C6-AHL and C8-AHL) with increased cell invasion at 37°C in vitro, and that this response most likely is sdiA mediated. These results indicate that if AHLs are present in the intestinal environment, they may increase the invasiveness of Salmonella.     

Nitrogen supply determines responses of yield and biomass partitioning of perennial ryegrass to elevated atmospheric carbon dioxide concentrations

Perennial ryegrass (Lolium perenne L. cv. Parcour) grown at eight levels of nitrogen (N) fertilization (0–765 mg/pot) was exposed to ambient (390 ppm) and elevated (690 ppm) carbon dioxide (CO₂) concentrations for 83 days. Plants were cut three times and dry matter yields determined for each harvest. At final harvest, dry weight of root and stubble biomass was determined, as N concentrations of all plant fractions were determined. Carbon dioxide enrichment effects on yield and total plant biomass increased with increasing N fertilization. The weaker CO₂‐related yield enhancement at low N supply was due to the plants inability to increase tiller number. Root fraction of total plant biomass at final harvest was increased by high CO₂ and decreased by N supply. Root biomass was significantly increased by CO₂ enrichment and for both CO₂ treatments the N supply for maximum root mass coincided with the N supply for reaching maximum total plant biomass. A significant correlation between root fraction of total plant dry matter and N concentration of total plant biomass, which was not changed by CO₂ enrichment, indicates that biomass partitioning between shoot and root is controlled by the internal N status of the plant.     

Effects of tenotomy of the tensor tympani muscle on the acoustic reflex in dogs

The acoustic reflex (AR) was recorded from 12 healthy mixed-breed dogs. Latency and amplitude were measured from ipsilateral and contralateral AR at stimulus frequencies of 1 and 2 kHz and intensities of 70 to 110 dB sound pressure level for ipsilateral AR and 70 to 120 dB hearing level for contralateral AR. Mean latencies for ipsilateral and contralateral AR were between 33.46 and 206.10 ms and between 45.26 and 180.89 ms, respectively, and amplitudes were between 0.14 and 1.79 cm3 and between 0.31 and 1.86 cm3 of air, respectively. Stimulus frequencies and intensities had significant effects (P less than 0.05) on ipsilateral and contralateral AR latencies and amplitudes. Ipsilateral and contralateral AR decays were determined by measuring compliance change during a 10-s pure-tone stimulation at frequencies of 1 and 2 kHz at an intensity of 10 dB above AR threshold. Reflex decays for 1 kHz and 2 kHz frequencies averaged 5.74% and 9.71%, respectively, for ipsilateral AR and 5.08% and 5.40%, respectively, for contralateral AR. Bilateral tympanograms and brain stem auditory-evoked responses were performed on each dog. Mean normal static compliance of the middle ear, as determined by tympanometry, was 0.15 cm3. Unilateral tenotomy of the tensor tympani muscle was done on 6 of the 12 dogs, and each of the preceding procedures were repeated within 1 week after surgical operation. Transection of the tensor tympani tendon did not alter (P greater than 0.05) the latencies or amplitudes of 1 kHz- or 2 kHz-evoked contralateral AR, the latency or amplitude of 1 kHz-evoked ipsilateral AR, or the amplitude of 2 kHz-evoked ipsilateral AR. However, the latency of 2 kHz-evoked ipsilateral AR was significantly (P less than 0.05) increased. Reflex decay increased significantly (P less than or equal to 0.001) for the contralateral reflex elicited by the 2 kHz stimulus. Neither compliance of the middle ear system nor amplitude and latency of the brain stem auditory-evoked response were affected (P greater than 0.05) by tenotomy. Since tenotomy eliminates participation of the tensor tympani in the AR, these data indicate that contraction of this muscle is not primarily responsible for the compliance changes recorded during an acoustic reflex in dogs.     

Prognostic factors for tumor remission and survival in dogs after surgery for primary lung tumor: 76 cases (1975-1985)

The association of various prognostic factors with remission and survival after the excision of lung tumors was evaluated in 76 dogs. Overall, the median survival time of treated dogs was 120 days; 72% had tumor that underwent remission (median duration of remission, 120 days). Dogs with tumors that underwent remission had significantly (P = 0.001) increased survival time (median, 330 days vs 28 days for dogs with tumors that did not undergo remission). The finding of normal-sized lymph nodes at the time of therapeutic thoracotomy was significantly (P = 0.001) correlated with increased remission probability (85.4% remission rate vs 43.6% in dogs with large lymph nodes). Use of various diagnostic methods to find normal regional lymph nodes before surgery indicated that such finding was significantly (P less than or equal to 0.01) correlated with increased remission duration (median remission duration, 365 days, vs 60 days for tumors in dogs with large lymph nodes), and the finding of normal lymph nodes at the time of surgery was significantly (P less than or equal to 0.01) correlated with increased survival time (median, 345 days, vs 60 days for dogs with large lymph nodes).     

Thoracic, Abdominal and Vertebral Actinomycosis

The clinical, laboratory, radiographic, and histologic features and the response to therapy in three dogs with actinomycosis are reported. One dog (dog 1) had a 12-cm nonresectable mass extending from the ventrolateral chest wall into the left ventricular myocardium. Another dog (dog 2) had a diffuse peritonitis with "sulfur granules" and two large masses. One of these masses was nonresectable involving adjacent abdominal structures. A third dog (dog 3) had a subvertebral mass at T1-3 producing quadraplegia. Two dogs had periosteal reactions involving adjacent sternebrae (dog 1) or ribs and vertebral bodies (dog 3) that are characteristic of Actinomyces spp infections. In dogs 1 and 2 the diagnosis was based on the morphologic and tinctorial properties of free sulfur granules and/or tissue granules. Culture results were variable. Tissue from dog 1 yielded no growth, while polymicrobial infections, which included Actinomyces spp, were identified in dogs 2 and 3. Actinomyces odontolyticus was isolated from dog 3. Although the actinomycotic granulomas were either not excised or only partially excised from dogs 1 and 2, both animals were cured by the oral administration of high doses of penicillin G for 19 and 6 months, respectively. Dog 3 responded dramatically to the same antibiotic therapy given for 5 months. However, within 4 months of discontinuing treatment an abscess and draining fistulous tracts developed in the left axillary region. Two surgical fistulectomies and additional penicillin therapy were required to cure this animal. These cases and the current veterinary and human literature on actinomycosis are used to propose a rational approach to the treatment of actinomycosis in the dog.