Distribution of latent bovine herpesvirus 2 DNA in tissues of experimentally infected sheep

The biology of latent infection by bovine herpesvirus 2 (BoHV-2), the agent of mammillitis in cows, remains largely unknown. We herein report attempts to reactivate the latent infection and investigated the sites of BoHV-2 latency in experimentally infected sheep. Ewes inoculated with BoHV-2 in the udder’s skin shed virus for up to five days, developed mammillitis and seroconverted. However, attempts to reactivate latent infection by dexamethasone administration at day 40 pi failed. Nevertheless, viral DNA - and not infectious virus - was detected by PCR in several nerve ganglia and/or regional lymph nodes (LNs) of all animals at day 40 post-reactivation. Likewise, lambs previously inoculated with BoHV-2 in the nose harbored latent viral DNA in trigeminal ganglia, tonsils and regional LNs. These results demonstrate that BoHV-2 establishes latent infection in nerve ganglia and in regional lymphoid tissues, yet virus reactivation is not easily achieved by standard protocols used.     

Effect of Steeping Conditions on Wet-Milling Characteristics of Hard Red Winter Wheat

A batch-wise small-scale wet-processing laboratory for whole wheat kernel has been designed and constructed to produce wheat starch and gluten from wheat grains. Hard red winter wheat kernels were steeped in three steeping media: SO₂ solution, lactic acid, and hydrochloric acid. Acid concentrations of 0.1, 0.3, and 0.5%, were used for SO₂ solutions and hydrochloric acid, and 0.1, 0.6, and 3.0% for lactic acid. After 16, 20, and 24 hr of steeping, the wheat was wet-milled. Yields and protein contents of wet-milling fractions were compared. Both high concentration of steeping media and long steeping time increased the starch yield and decreased the protein contents of the starch. However, the steeping time and acid concentration could be reduced from 24 to 20 hr and from 0.5 to 0.3%, respectively, without any statistically significant difference in starch yields or protein contents of the starch. Consistency and color of the starch were affected by both steeping time and acid concentrations of steeping media.     

Characterization ofSolanum tuberosum simple sequence repeats and application to potato culiwar identification

With the continued introduction of new potato cultivars, accurate identification is becoming difficult but is essential for maintaining cultivar integrity and Plant Breeders’ Rights. Hypervariable DNA sequences, referred to as simple sequence repeats (SSRs) or microsatellites, have been reported to be an excellent source of genetic markers. To determine the abundance, distribution, and composition of SSRs withinSolanium tuberosum, 252 sequences were searched for tetranucleotide and smaller SSRs with a minimum length of 20 nucleotides and a maximum discrepancy of two nucleotides. In total, 40 unique SSRs were observed in the 252S. tuberosum sequences examined and occurred at a frequency of one SSR every 8.1 kb. To assess the ability of site-specific amplified SSRs to identify potato cultivars, a simple (TCAC)_m and compound (TCAC)_m ? (CTT)_n SSR 5’ to the starch synthase gene and a compound (C)_p ? (CT)_q ? (AT)_r ? (G)_s SSR 5’ to the sequence encoding mature proteinase inhibitor I, were examined and shown to produce unique DNA profiles for 73 of 95 tetraploid cultivars. In total, 24 alleles were observed at these loci and the accurately sized amplified DNA products can be used to establish a database for cultivar identification. Site-specific amplified alleles were somatically stable and have been conserved in clonal variants of Russet Burbank independently maintained for almost seven decades, a characteristic essential for cultivar identification. As genetic markers, the abundant, informative, and easily examined site-specific amplified alleles of SSRs are ideal for quickly and accurately determining cultivar identity of S.tuberosum ssp.tuberosum.     

Wound contamination and antimicrobial susceptibility of bacteria cultured during total ear canal ablation and lateral bulla osteotomy in dogs.

OBJECTIVE: To detect contamination of wound sites from surgical handling of excised tissues during total ear canal ablation and lateral bulla osteotomy in dogs, and to compare susceptibility of bacterial isolates to cefazolin with susceptibility to other antimicrobial agents. DESIGN: Prospective clinical study. ANIMALS: 13 dogs. PROCEDURE: Dogs were treated surgically for otitis externa and media via total ear canal ablation and lateral bulla osteotomy. Specimens for aerobic bacterial culture were obtained from SC tissue immediately following skin incision, tissues excised from the osseous bulla (after transection of the horizontal ear canal and lateral bulla osteotomy), and from SC tissue prior to skin closure. Antimicrobial susceptibility of bacterial isolates to various antibiotics was determined by use of a broth dilution assay. RESULTS: There was a significant association between isolation of Streptococcus canis and Escherichia coli from specimens from the osseous bulla and specimens from the SC tissues prior to skin closure, indicating contamination of the SC tissues during surgery. Seventy percent of bacterial isolates were susceptible to cefazolin. CLINICAL IMPLICATIONS: Measures to limit bacterial contamination resulting from tissue handling during total ear canal ablation and lateral bulla osteotomy are necessary. Bacteriologic culture of tissue of the osseous bulla and determination of antimicrobial susceptibility are recommended. Administration of cefazolin alone may not be efficacious for antimicrobial prophylaxis.     

Rhabdochona longleyi sp. n. (Nematoda: Rhabdochonidae) from blind catfishes, Trogloglanis pattersoni and Satan eurystomus (Ictaluridae) from the subterranean waters of Texas

A new nematode species, Rhabdochona longleyi sp. n. is described from the intestine of two species of blind catfishes, Trogloglanis pattersoni Eigenmann (type host) and Satan eurystomus Hubbs et Bailey (both fam. Ictaluridae, Siluriformes) from the subterranean waters (artesian wells penetrating San Antonio pool of Edwards Aquifer) of Texas, USA. It is characterized largely by the presence of only six anterior teeth in the prostom, simple deirids, by the shape and length of spicules (0.42 to 0.50 mm and 0.093-0.102 mm), shape of the tail tip (rounded), and by filamented eggs. R. longleyi probably adapted to the environment of the aquifer by utilizing available troglobitic crustaceans instead of aquatic insects as an intermediate host.     

Induction of estrus in greyhound bitches with prolonged idiopathic anestrus or with suppression of estrus after testosterone administration

Twelve anestrous adult Greyhound bitches were used to study a regimen for induction of estrus. Once daily, 7 bitches were given diethylstilbestrol (DES; 5 mg, PO) until sanguineous vaginal discharge and vulvar edema were observed (designated as day 1 of proestrus) and for 2 days thereafter. If no response was elicited after 7 days, a doubled DES dose was given for up to an additional 7 days. Luteinizing hormone (5 mg, IM) was given on day 5 of proestrus, and follicle-stimulating hormone (10 mg, IM) was given on days 9 and 11 of proestrus. Bitches were bred once on day 13. Five bitches were used as a control group; they were given candy tablets for 7 days (first day on tablets, treatment day 1) and 0.9% NaCl (1.0 ml, IM) on treatment days 12, 16, and 18. The 7 bitches treated with DES had a mean proestrus period of 7.7 days and a mean estrus period of 5.7 days up to the day of mating. After mating, they had a mean gestation interval of 64 days and delivered a mean of 4 pups/litter. In 5 bitches, initial treatment with 5 mg of DES/day induced proestrus within 7 days; however, in 2 bitches, additional treatment with 10 mg of DES/day was needed for 5 and 6 days, respectively. Serum estradiol-17 beta and progesterone concentrations remained at base line during the period of DES treatment. Concentrations of both hormones increased after injection with luteinizing hormone and remained high for the next 4 days.(ABSTRACT TRUNCATED AT 250 WORDS)     

Molecular cloning and expression of kidney‐type glutaminase from common carp (Cyprinus carpio) and its up‐regulation by glutamine in primary culture enterocyte

Glutaminase (GLS) is the key enzyme of glutamine (Gln) metabolism and utilization. In this study, a cDNA encoding GLS protein was identified from common carp Cyprinus carpio intestine. The open reading frame of GLS cDNA encodes a polypeptide of 595 amino acids, which shows a high similarity with its zebrafish Danio rerio counterpart. Bioinformatic analysis showed the protein belongs to kidney‐type GLS. The putative protein has glutaminase domain and ankyrin repeats domain, which are highly conserved among vertebrate orthologues. Real‐time quantitative PCR analysis revealed that the abundance of GLS mRNA was the highest in the white muscle, followed by the brain, eyeball and pituitary. Glutaminase was ubiquitously expressed in all intestinal segments of common carp. The activity of GLS did not distribute uniformly along the entire length of the intestine. In primary culture enterocyte, and the expression of GLS mRNA is up‐regulated quickly and effectively by Gln.