A novel phenotype for Laron dwarfism in miniature Bos indicus cattle suggests that the expression of growth hormone receptor 1A in liver is required for normal growth

Mutations within the growth hormone receptor (GHR) gene that lead to an inactivated or truncated GHR protein cause abnormal growth and small adult size in a variety of species (Laron dwarfism). We studied a line of miniature Bos indicus cattle that have phenotypic (small mature size) and endocrine (increased blood growth hormone and decreased blood insulin-like growth factor-I concentrations) similarities to Laron dwarfs. Liver mRNA from miniature and control cattle was used to amplify a cDNA within the coding region of the GHR. The miniature cattle had GHR mRNA size (determined by Northern blot) and cDNA sequence that were similar to control cattle and, therefore, were unlike most Laron dwarf genotypes in which the GHR gene is mutated. Amounts of mRNA from liver as well as muscle (superficial neck and longissimus) were analyzed by ribonuclease protection assay for IGF-I, total GHR, GHR 1A (inducible, liver-specific GHR mRNA), and GHR 1B (constitutive GHR mRNA). Four control and five miniature bulls were tested. As expected, liver IGF-I mRNA was decreased in the miniature cattle (approximately 12% of control; P < 0.01). The amount of the total GHR as well as GHR 1A mRNA were also decreased in liver (17% and 19% of control, respectively; P < 0.01). Other GHR mRNA, including GHR 1B mRNA, were similar for miniature and control cattle. In muscle, there was a tendency (P < 0.10) for decreased IGF-I mRNA and increased GHR mRNA in miniature compared with control cattle. In summary, a novel phenotype for Laron dwarfism in Bos indicus cattle was associated with underexpression of GHR 1A mRNA, but not other GHR mRNA variants in liver. In addition to decreased GHR 1A mRNA, the miniature cattle had decreased liver IGF-I mRNA. Full expression of GHR 1A in liver, therefore, may be required for full liver IGF-I expression and normal growth.     

Intramyocardial haematoma causing right ventricular outflow obstruction after brown snake (Pseudonaja species) envenomation in a dog

A 15-month-old, male neutered Staffordshire Bull Terrier cross was presented to its referring veterinarian collapsed and agonal. He was immediately intubated, manually ventilated, and treatment commenced for presumptive snake envenomation with two vials of Tiger/Multi-Brown Snake Antivenom (minimum 7000 units/vial). The dog was transferred to a referral hospital intubated. Additional diagnostics performed following arrival at the referral hospital included a urine snake venom detection kit test, which was positive for brown snake immunotype. Three additional vials of Tiger/Multi-Brown Snake Antivenom (minimum 7000 units/vial) were administered until the dog was extubated and able to stand. Venom-induced consumptive coagulopathy (VICC) was diagnosed based on prolonged clotting times and scleral haemorrhage. Paroxysms of right ventricular outflow tract (RVOT) origin ventricular arrhythmias were treated with lignocaine and sotalol. Four days after presentation, a new-grade IV/VI systolic heart murmur was auscultated, prompting an echocardiogram. An anechoic and compartmentalised mass measuring 43 mm × 19 mm was visualized within the right ventricular wall at the RVOT, immediately adjacent to the pulmonic valve. The mass was causing a RVOT obstruction. Its appearance was suggestive of an intramyocardial haematoma, most likely secondary to VICC. The dog remained cardiovascularly stable, and treatment consisted of supportive care. Recheck echocardiograms at 2 and 7 weeks after discharge revealed progressive improvement of the intramyocardial mass and resolution of the associated heart murmur. Although intramyocardial haematomas are rare, it should be considered as a differential in dogs that develop a newly diagnosed heart murmur and/or cardiac arrhythmia following brown snake envenomation.     

Report on the analyses of mAb reactive with porcine CD8 for the second international swine CD workshop

Based on an analysis of their reactivity with porcine peripheral blood lymphocytes (PBL), only three of the 57 mAbs assigned to the T cell/activation marker group were grouped into cluster T9 along with the two wCD8 workshop standard mAbs 76-2-11 (CD8a) and 11/295/33 (CD8b). Their placement was verified through the use of two-color cytofluorometry which established that all three mAbs (STH101, #090; UCP1H12-2, #139; and PG164A, #051) bind exclusively to CD8⁺ cells. Moreover, like the CD8 standard mAbs, these three mAbs reacted with two proteins with a MW of 33 and 35 kDa from lymphocyte lysates and were, thus, given the wCD8 designation. Because the mAb STH101 inhibited the binding of mAb 76-2-11 but not of 11/295/33, it was given the wCD8a designation. The reactivity of the other two new mAbs in the T9 cluster with the various subsets of CD8⁺ lymphocytes were distinct from that of the other members in this cluster including the standards. Although the characteristic porcine CD8 staining pattern consisting of CD8^low and CD8^high cells was obtained with the mAb UCP1H12-2, a wider gap between the fluorescence intensity of the CD8^low and CD8^high lymphocytes was observed. In contrast, the mAb PG164A, not only exclusively reacted with CD4⁻/CD8^high lymphocytes, but it also failed to recognize CD4/CD8 double positive lymphocytes. It was concluded that this mAb is specific for a previously unrecognized CD8 epitope, and was, thus, given the wCD8c designation. A very similar reactivity pattern to that of PG164A was observed for two other mAbs (STH106, #094; and SwNL554.1, #009). Although these two mAbs were not originally positioned in the T cell subgroup because of their reactivity and their ability to inhibit the binding of PG164A, they were given the wCD8c designation. Overall, five new wCD8 mAbs were identified. Although the molecular basis for the differences in PBL recognition by these mAbs is not yet understood, they will be important in defining the role of CD8⁺ lymphocyte subsets in health and disease.     

Evaluation of biomarkers of kidney injury following 4% succinylated gelatin and 6% hydroxyethyl starch 130/0.4 administration in a canine hemorrhagic shock model

Objective : To investigate the association between synthetic colloids and biomarkers of acute kidney injury (AKI) in dogs with hemorrhagic shock. Design : Experimental interventional study. Setting : University. Animals : Twenty‐four healthy ex‐racing Greyhounds. Interventions : Anesthetized Greyhounds subjected to hemorrhage for 60 min were resuscitated with 20 mL/kg of fresh whole blood (FWB), 6% hydroxyethyl starch (HES) 130/0.4, 4% succinylated gelatin (GELO), or 80 mL/kg of isotonic crystalloid (CRYST) over 20 min (n = 6 per treatment). Concentrations of biomarkers of AKI were measured at baseline, end of hemorrhage, and at 40 (T60), 100 (T120), and 160 (T180) min after fluid bolus. Biomarkers included neutrophil gelatinase‐associated lipocalin in urine and serum (uNGAL; sNGAL), and urine cystatin C (uCYSC), kidney injury molecule‐1 (uKIM), clusterin (uCLUST), osteopontin, gamma‐glutamyl transferase, monocyte chemoattractant protein‐1 (uMCP), interleukin‐6, interleukin‐8, protein (uPROT), hyaluronan, and F₂‐isoprostanes. Renal histology was scored for tubular injury and microvesiculation. Biomarker fold‐change from baseline was compared between groups using mixed effects models (Bonferroni–Holm corrected P<0.05). Frequencies of histology scores were compared by Fisher's exact test. Measurements and main results : In dogs treated with GELO, uNGAL fold‐change was markedly greater compared with all other groups at T60, T120, and T180 (all P<0.001), and uCYSC was greater at T60 compared with CRYST (P<0.001), and at T120 and T180 compared with all other groups (all P<0.001). Smaller, albeit significant, between‐group differences in uKIM, uCLUST, uMCP, and urine protein concentration were observed across the FWB, GELO, and HES groups, compared with CRYST. The GELO group more frequently had marked tubular microvesiculation than the other groups (P = 0.015) although tubular injury scores were comparable. Conclusion : In dogs with hemorrhagic shock, GELO was associated with greater magnitude increases in urine biomarkers of AKI and more frequent marked tubular microvesiculation, compared with FWB, CRYST, and HES.     

Restoring the Sagebrush Component in Crested Wheatgrass–Dominated Communities

Monotypic stands of crested wheatgrass (Agropyron cristatum [L] Gaertm. and Agropyron desertorum [Fisch.] Schult.), an introduced grass, occupy vast expanses of the sagebrush steppe. Efforts to improve habitat for sagebrush-associated wildlife by establishing a diverse community of native vegetation in crested wheatgrass stands have largely failed. Instead of concentrating on a diversity of species, we evaluated the potential to restore the foundation species, Wyoming big sagebrush (Artemisia tridentata spp. wyomingensis [Beetle & A. Young] S. L. Welsh), to these communities. We investigated the establishment of Wyoming big sagebrush into six crested wheatgrass stands (sites) by broadcast seeding and planting seedling sagebrush across varying levels of crested wheatgrass control with glyphosate. Planted sagebrush seedlings survived at high rates (～ 70% planted sagebrush survival 3 yr postplanting), even without crested wheatgrass control. However, most attempts to establish sagebrush by broadcast seeding failed. Only at high levels of crested wheatgrass control did a few sagebrush plants establish from broadcasted seed. Sagebrush density and cover were greater with planting seedlings than broadcast seeding. Sagebrush cover, height, and canopy area were greater at higher levels of crested wheatgrass control. High levels of crested wheatgrass control also created an opportunity for exotic annuals to increase. Crested wheatgrass rapidly recovered after glyphosate control treatments, which suggests multiple treatments may be needed to effectively control crested wheatgrass. Our results suggest that planting sagebrush seedlings can structurally diversify monotypic crested wheatgrass stands to provide habitat for sagebrush-associated wildlife. Though this is not the full diversity of native functional groups representative of the sagebrush steppe, it is a substantial improvement over other efforts that have largely failed to alter these plant communities. We also hypothesize that planting sagebrush seedlings in patches or strips may provide a relatively inexpensive method to facilitate sagebrush recovery across vast landscapes where sagebrush has been lost.     

The significance of small vs large intestinal digestion of cereal grain and oilseed protein in the equine

Mature ponies fitted with permanent ileal cannulas were used in two 3×3 Latin square experiments to quantify prececal, postileal and total tract digestion of N. In trial 1, corn, oats and sorghum were each fed with coastal Bermuda grass hay in 75:25 ratios. Apparent prececal digestibilities were similar (P>.05) and averaged 46.6%. By-difference procedures were employed to calculate digestibility of the cereal grain N only and apparent prececal N digestibility averaged 57.1%. In trial 2, a basal corn and hay diet was supplemented with cottonseed meal and soybean meal. Apparent total tract N digestibilities were similar (P>.05) across treatments, and prececal digestibility averaged 45.6%. By-difference calculations were used to determine digestibility of SBM and CSM N alone. Apparent prececal digestibility of SBM was 52.5% and was lower (P<.05) than 81.2% for CSM. It is possible that inadequate or excessive heat treatment of SBM affected enzymatic digestion. True digestibility of total rations fed in trial 2 was estimated by linear regression of nitrogen digested on nitrogen intake or N presented to the large intestine. True N digestibility of diets containing SBM and CSM was 54.7% and 69.4%, respectively.     

Poind Soil pH Measurement

Soil pH often is measured in samples from the bottoms of aquaculture ponds. Several different techniques for soil pH are used. This study considered the differences in pH obtained by the different methods and determined which methods appeared most useful. Dual electrodes (indicating and reference) and a single‐probe combination electrode gave similar pH values when inserted into 1:1 mixtures of dry soil and distilled water. There were slight differences in pH between readings with dual and combination electrodes when the dual electrodes were arranged with the indicating electrode in the sediment phase and the reference electrode in the supernatant phase of the mixture. The two‐phase method with the dual electrode does not appear warranted because of greater difficulty in making measurements. Dry soil: distilled water ratios of 1:2.5, 1:5, and 1:10 had progressively greater pH readings than obtained at a 1:1 ratio. Measurements made in 0.01 M CaCl₂ and 1.0 M KCl had much different values than those made in distilled water. Higher pH resulted when pH was measured without stirring or in filtrates of soil‐water mixtures. A 20‐min period of intermittent stirring before making measurements was necessary for a stable pH value. Particle size did not influence pH in aliquots passing 0.053 to 2.36‐mm sieves. Drying temperature had a strong influence on pH, and measurements made on samples dried at 40 to 60 C are probably most reliable. Measurements of in situ pH in wet soil with standard pH electrode or a portable acidity tester differed greatly from those made in 1:1 dry soil to distilled water mixtures. Pond bottom soil pH measurement should be standardized. Based on findings of this study, the following method is suggested: dry soil at 60 C in a forced‐draft oven; pulverize soil to pass a 2‐mm sieve; mix soil and distilled water in a 1:1 ratio (weight: volume); stir intermittently with glass rod for 30 min; insert dual electrodes or a combination electrode into the mixture; measure pH while stirring.