Analysis of amino acid uptake and translocation in Arabidopsis with a low‐cost hydroponic system

In soils, amino acids may be an important source of nitrogen for plants, at least in those where organic matter is not quickly degraded. The physiology of uptake of amino acids by roots was mainly studied in the 70's and 80's, before genes encoding amino acid importers were cloned in the 90's. While two families of amino acid transporters have been identified, yielding a total of about 100 genes, the role of each member is yet to be elucidated. As a tool for studying the role of amino acid transporters from Arabidopsis we set up a new hydroponic system suitable for radioisotope use. This system enables reproducible amino acid uptake by roots and estimation of the transport to the shoots of the amino acid taken up. We show that the rates of glutamine (Gln) uptake by wild‐type roots and transfer to the shoots were linear, and that other tested amino acids were translocated to the shoots with lower efficiency than Gln. A T‐DNA insertion mutant for a Gln exporter was compared to the wild‐type plants. Gln uptake and transfer were similar in both genotypes, showing that the suppression of the exporter did not affect uptake or transfer of amino acids to the shoots. The main advantage of the hydroponic system presented here is that all the materials used to grow Arabidopsis are virtually free and can therefore be discarded, a useful feature when working with radioactivity.     

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.     

Organic Carbon and Dissolved Oxygen Budgets for a Commercial‐Size,In‐pond Raceway System

Intensive production of ictalurid catfish in the USA has increased over the past several years, and a better understanding of the amount of organic carbon (OC) and dissolved oxygen (DO) in these culture environments is needed. Budgets for OC and DO were estimated over a production season (March to November) for an in‐pond raceway system for channel catfish, Ictalurus punctatus, and hybrid catfish (channel catfish × blue catfish, Ictalurus furcatus), with co‐culture of paddlefish, Polyodon spathula, and Nile tilapia, Oreochromis niloticus. Feed input ranged from 45 to 263 kg/ha/d with an overall average of 80 kg/ha/d. Production of each kilogram of live catfish required 1.5 kg of feed and released into the water 0.70 kg of OC, and led to the synthesis of an additional 3.36 kg of OC by photosynthesis. Consequently, production of 1 kg of live catfish resulted in 4.06 kg OC, and harvest of catfish accounted for only 29.5% of OC applied from the feed. Removal of OC increased to 34.3% with the additional harvest of paddlefish and tilapia. OC was consumed in respiration, and some OC accumulated in sediment. Total respiration within the system exceeded the DO produced by photosynthesis, while diffusion and mechanical aeration aided in maintaining suitable DO levels for fish production.     

Embodied Resources in Fish and Shrimp Feeds

Global averages were obtained for amounts of energy, land, water, wildfish, nitrogen, and phosphorus embodied in aquaculture feed ingredients. These data allowed amounts of these embodied resources to be calculated for typical feed formulations for channel catfish, Ictalurus punctatus; hybrid catfish, I. punctatus♀ × I. furcatus♂; Vietnamese catfish, Pangasius spp.; Atlantic salmon, Salmo salar; rainbow trout, Oncorhynchus mykiss; tilapia, Oreochromis spp.; whiteleg shrimp, Litopenaeus vannamei; and black tiger shrimp, Penaeus monodon. Embodied resource use per m.t. of feed varied among species: energy, 4.90–12.48 GJ/m.t.; land, 0.082–0.312 ha/m.t.; water, 502–1227 m³/m.t.; wildfish, 0–2880 kg/m.t.; nitrogen, 3.08–8.63 kg/m.t.; phosphorus, 1.16–5.62 kg/m.t. These calculations did not account for variations in site‐specific factors related to embodied resources and feed composition and use. But they suggest that reducing feed conversion ratio (FCR) by 0.1 unit for the seven species (species groups) could potentially reduce feed use by around 1.1 million tonne (Mt) while conserving 9.8 million GJ of energy, 270,000 ha of agricultural land, 1.4 billion m³ of freshwater, and 1.24 Mt of wildfish. Reduction of the FCR is a powerful means of lessening farm‐level production costs and negative impacts of feed production and use.     

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 the Ultraviolet Spectrophotometric Method for the Measurement of Total Nitrogen in Water

Abstract.— A simple, accurate, and environmentally sound procedure based on spectroscopy in the UV range was modified to measure total nitrogen concentrations in both freshwater and brackishwater. Water samples from freshwater fish ponds were analyzed for total nitrogen with the persulfate digestion method followed by analysis of nitrate with the UV method for comparison with the NAS reagent method. Brackishwater samples with salinities of 8 to 10 ppt from shrimp ponds were digested by the persulfate method and analyzed with the UV method. Percentage recovery of nitrogen and also precision were evaluated for the UV method. Nitrate concentrations in the freshwater persulfate digests determined by the UV method were similar to those obtained by the NAS reagent method ( P < 0.05). The slope of the regression line did not differ from 1.0 ( P < 0.05). Precision of both methods was acceptable over the range of nitrogen concentration used, but precision of the UV method tended to be better than that of the NAS method. Spike recovery averaged 97.4% in the NAS reagent method and 107.2% in the UV method. Interference from organic matter was minor in the UV method, because virtually all of the organic matter was oxidized during the digestion. Precision with brackishwater samples was slightly better than the freshwater samples and percentage spike recovery averaged 102.8%. Because spectrophotometers in most laboratories have the capability of reading in the UV range, the UV method is an attractive alternative for total nitrogen analysis.     

Habitat quality,not habitat amount,drives mammalian habitat use in the Brazilian Pantanal

Landscape Ecology - An understanding of species-habitat relationships is required to assess the impacts of habitat fragmentation and degradation. To date, habitat modeling in fragmented landscapes...     

Bottom soil quality in Tilapia ponds of different age in Thailand

Bottom soil samples were collected from 35 ponds in the vicinity of Samutprakarn, Thailand. Ponds ranged in age from 3 to 39 years and had been used continuously for production of tilapia. Liming materials had been applied in large amounts, and bottom soils of all ponds had pH above 7, low exchange acidity, and free carbonate. Pond soils often contained between 1% and 2% total sulphur, suggesting that they were potential acid–sulphate soils. However, acidity from sulphide oxidation was not expressed because carbonate in the soil neutralized it. Concentrations of total carbon seldom exceeded 4% and the average for organic carbon was 1.90%. The correlations between pond age and both total carbon and organic carbon concentration were weak (r=0.34 and 0.36 respectively). Concentrations of nitrogen in bottom soils did not differ with pond age and ranged from 0.1% to 0.3% with an average of 0.19%. The average ratio of concentrations of carbon and nitrogen was 11. Acid‐extractable phosphorus concentrations averaged 217 mg kg^?1, but the phosphorus adsorption capacity averaged 768 mg kg^?1 suggesting that soils still have considerable reserve capacity to adsorb phosphorus. Ponds can be used annually for semi‐intensive production of tilapia, and presumably other species, for many years without serious deterioration of bottom soil quality.