Contribution of beta-lactamase production to the resistance of mycobacteria to beta-lactam antibiotics

Changes in motor function were assessed in male rats after injecting graded doses (100, 200, 400, and 800 mg/kg, IP) of ammonium chloride and ammonium acetate. The effects were correlated with the concentrations of ammonia and glucose in the brain and blood. Spontaneous motor activity and motor coordination were inhibited after injecting 100 and 200 mg/kg, whereas with 400 and 800 mg/kg the animals exhibited convulsive movements. A dose-dependent increase was found in the concentrations of ammonia and glucose in both blood and brain. These were restored, 25 min after treatment, to control levels in the blood and not in the brain. A correlation was found between the time courses of inhibitory motor events and a rise in brain ammonia levels. Convulsant action of ammonium salts was accompanied by a marked elevation of ammonia and glucose concentration in the brain. The findings suggest that detoxication of diffused ammonia is a rate-limiting process in the brain and that ammonia, at toxic concentrations, decreases glucose utilization in the brain, resulting in an inhibition of motor function. A very high concentration of ammonia in the brain, although inhibiting glucose utilization, produces clonic convulsions probably by activating directly the motor neurons.     

The major excretory/secretory protease from Lucilia cuprina larvae is also a gut digestive protease

The larvae of the fly Lucilia cuprina excrete or secrete a chymotrypsia (LCTb) onto the skin of sheep to facilitate the establishment of the larval infestation. A combination of immunoblotting and RT-PCR approaches has established that this protease is also a gut digestive protease. LCTb is synthesized primarily in the cardia, a small highly specialized organ located at the anterior end of the midgut and by midgut cells. There is also some expression by the hindgut but no expression by salivary glands. Excretion of LCTb with waste products or regurgitation of the gut contents of the larvae may explain how this protease is transferred from the larval gut onto ovine skin. LCTb is first expressed in eggs and constitutively expressed throughout each larval instar, but is not expressed in pupae or adult flies. It is concluded that LCTb could be involved in the establishment of larvae on sheep skin as well as acting as a general gut digestive enzyme.     

Kinetic disposition of an emulsifiable concentrate formulation of deltamethrin applied to sheep in a plunge-dip and its effect on lice

An emulsifiable concentrate formulation of the synthetic pyrethroid insecticide deltamethrin was applied in a plunge dip, 3 weeks after shearing, to a group of 5 Merino sheep infested with sheep body lice, Bovicola ovis. Deltamethrin concentrations on the wool were measured at regular intervals between 1 and 98 days after treatment and were not significantly different (P > 0.05) between sites on the dorsal mid-line, upper or lower flank. Levels in the tip of the fleece were significantly greater than those in the base, indicating that there was little movement of deltamethrin down the staple as the wool grew. Most lice were killed after 20 h of exposure in vitro to wool samples collected between 1 and 28 days after treatment. However, many lice survived in samples containing a similar concentration of deltamethrin, but collected between 35 and 98 days after treatment. Numbers of lice surviving increased with the sampling time after treatment, suggesting that the bioavailability of the deltamethrin changed as the insecticide aged in the fleece. Some transfer of deltamethrin occurred from treated to untreated sheep. The levels of deltamethrin were higher in sheep placed in contact with the treated group at 14 days after treatment than in those which were in contact from 43 days after treatment.     

Pruritus and dermal response to insect antigens in sheep infested with Bovicola ovis

This study examined the relationships among louse density, pruritus and dermal response to insect antigens in sheep infested with Bovicola ovis. Polypay and Columbia ewes were allocated to two groups, infested and naive, and louse densities and pruritus were monitored for 15 months. Ten months after the initial infestation, all sheep were tested for hypersensitivity on the midside and ears by intradermal injection of soluble extracts of B. ovis, Stomoxys calcitrans and Musca autumnalis. The areas of skin reactions were measured at 20 min, 1, 3 and 24 h after injection and skin thickness was measured at 24 h. Louse densities on Polypays were approximately 10 times greater than on Columbias, and pruritus was correlated with louse numbers at most inspections. Most pruritic behaviour was directed to the sides of infested sheep. Wheal and flare reactions developed rapidly to all extracts in both infested and naive ewes. Reactions to louse extract were larger in infested than naive sheep at all four times after injection. In the infested Polypays, reactions to louse extract were greater than to the fly extracts, but in naive sheep there was little difference among extracts. Reactions in naive Columbias were larger than in naive Polypays at 20 min, 1 and 3 h, but had almost completely abated in both groups at 24 h. Reactions in infested Columbias were greater than in infested Polypays at 20 min, but at 24 h reactions in the Polypays were larger. Louse numbers and pruritus were correlated with wheal areas and skin thickness at 24 h, but there was little relationship with the size of reactions at earlier times. These findings are consistent with the development of a hypersensitive response to B. ovis and suggest that dermal reactions to lice may influence sheep susceptibility.     

Preliminary studies on humoral immune response of sheep to wohlfahrtiosis

The humoral immune response of sheep to wohlfahrtiosis was studied. An enzyme-linked immunosorbent assay was developed to compare four different types of antigens obtained from the third-stage larvae of Wohlfahrtia magnifica. The antigen prepared from salivary glands detected a humoral response in all 35 infested sheep and was more specific in the ELISA than cuticular, intestinal or whole larval antigens. The level of the humoral response in sheep to wohlfahrtiosis differed according to the location of the wounds.     

Effect of level of dietary energy and protein on embryo survival and progesterone production on day eight of pregnancy in Rasa Aragonesa ewes

The aim of this experiment was to investigate the effects of dietary protein and energy on ovulation rate and embryo survival to day 8 of pregnancy, and the associated concentrations of progesterone in jugular, ovarian and uterine veins, in a Spanish breed of sheep. In mid-October, three groups of ewes were fed to provide 1.5 x (H; n = 9), 0.5 x (L; n = 12) or 0.5 x plus 7.44 g CP/MJ ME (LP; n = 8) energy requirements for maintenance of live weight from day -14 relative to a synchronized mating on day 0. A significant effect of nutrition on ovulation rate was observed (H: 2.22 +/- 0.16; L: 1.50 +/- 0.16; LP: 1.88 +/- 0.12 corpora lutea; P < 0.05). Mean LH and progesterone concentrations were affected by nutrition on day 7, L ewes showing the highest mean LH level (P < 0.01), while H ewes presented the lowest mean LH concentration and the highest mean plasma progesterone concentration (P < 0.01). Laparotomies were performed on six animals of each group on day 8 to determine the effect of nutrition on embryo development. A significantly higher percentage of embryos recovered from L and LP ewes presented an earlier stage of development (morulae or early blastocysts) (P < 0.001), while 100% embryos of H ewes were expanded blastocysts. The ratio expanded blastocysts/corpora lutea was significantly higher in H ewes (0.86) when compared with L and LP groups together (0.57; P < 0.05). Mean progesterone concentration in the ovarian vein was 800-fold higher than mean jugular venous levels with no differences between groups. Samples from ovarian veins contralateral to corpus luteum-bearing ovaries showed mean progesterone concentrations significantly lower than samples opposite to corpus luteum (ipsilateral: 1037.84 +/- 138.45; contralateral: 30.4 +/- 11.22 ng/ml; P < 0.001). Mean progesterone concentration in the uterine vein was approximately 30-fold higher than in jugular and similar in both uterine horns and treatments. No effect of nutrition on pregnancy rate was observed (H: 89%; L: 92%; LP: 100%). These results suggest that neither dietary energy nor protein are able to modify pregnancy rate or progesterone concentrations in ovarian and uterine veins eight days after mating. However, the delay in embryo development observed in the embryos collected from L and LP ewes may give rise to compromised embryo growth and development some days later.     

Dissolution kinetics of malachite in ammonia/ammonium carbonate leaching

The leaching of oxide copper ore containing malachite, which is the unique copper mineral in the ore, by aqueous ammonia solution has been studied. The effect of leaching time, ammonium hydroxide, and ammonium carbonate concentration, pH, [NH₃]/[NH₄⁺] ratio, stirring speed, solid/liquid ratio, particle size, and temperature were investigated. The main important parameters in ammonia leaching of malachite ore are determined as leaching time, ammonia/ammonium concentration ratio, pH, solid/liquid ratio, leaching temperature, and particle size. Optimum leaching conditions from malachite ore by ammonia/ammonium carbonate solution are found as ammonia/ammonium carbonate concentrations: 5 M NH₄OH+0.3 M (NH₄)₂CO₃; solid/liquid ratio: 1:10 g/mL; leaching times: 120 min; stirring speed: 300 rpm; leaching temperature: 25 °C; particle size finer than 450 μm. More than 98% of copper was effectively recovered. During the leaching, copper dissolves as in the form of Cu(NH₃)₄⁺² complex ion, whereas gangue minerals do not react with ammonia. It was determined that interface transfer and diffusion across the product layer control the leaching process. The activation energy for dissolution was found to be 15 kJ mol⁻¹.     

Clinical, clinico-pathological and serological studies of Babesia ovis in experimentally infected sheep

Clinical, clinico-pathological and serological studies were performed in sheep experimentally infected with Babesia ovis. Acute babesiosis occurred in all the lambs infested with adult Rhipicephalus bursa ticks and in one lamb infested with the larvae. The rate of parasitaemia and the degree of anaemia were not correlated. Decrease in the packed-cell volume ranged from 30 to 40%. Parasitized erythrocytes were not observed to block capillaries in the brain, which explained the absence of nervous symptoms in acute babesiosis. The kidneys were the most severely affected organs, exhibiting acute glomerulonephritis. The lesions observed were suggestive of vascular alteration and vascular stasis, leading to anoxia of the tissues. A disseminated intravascular coagulation (DIC) syndrome was recorded in sheep infected with babesiosis. A marked increase in the enzymes of the transaminase groups, mainly aspartate aminotransferase (AST), was observed. Enzymatic changes (increases in AST, alanine aminotransferase (ALT) and lactic dehydrogenase (LDH) and decreases in sorbitol dehydrogenase (SDH), alkaline phosphatase (ALP) and malic enzyme (MEZ)), decreases in total proteins and albumin, and increases in urea and creatinine might reflect the degree of severity of the damage to the liver and kidney tissues. Most of the lambs (85%) that were infested with larvae, and all lambs infested with adult R. bursa ticks, reacted serologically to B. ovis antigen. The serological reactions following infestation with the larvae occurred much later than those following infestation with the adult stage. The lambs which were infested with larvae showed mild clinical reactions when challenged by infected R. bursa adults, as compared with the reactions to the challenge in naive control animals. The serological findings, in addition to the fact that one splenectomized lamb reacted to larval infestation with acute ovine babesiosis, show that the preimaginal stages of R. bursa can transmit B. ovis, usually causing a sub-clinical disease. It is suggested that infections derived from preimaginal ticks in the winter can preimmunize sheep for the subsequent more severe infections derived from adult ticks in the summer. Furthermore, in the absence of a reliable vaccine against B. ovis, grazing flocks in the enzootic regions should be exposed to the preimaginal stages during their activity period (October-February) before exposure to the adult ticks in spring and summer (April-July).     

Detection of sequential genetic alterations relevant for breast cancer development

Infusion of LPS or nematode larvae into the mammary glands of sheep induces recruitment of neutrophils or eosinophils respectively. While neutrophil recruitment required only a single infusion of LPS, repeated infusions of parasite larvae were required to induce significant eosinophil migration into the lumen of the glands. Eosinophil recruitment was accompanied by a distinct population of lymphocytes consisting mainly of activated (MHC class II and CD25+) T cells. L-selectin was expressed at reduced levels on both neutrophils and eosinophils collected from the mammary gland compared with cells present in the blood of the same sheep. In addition, VLA-4 and beta 1-integrin were down-regulated or negative in mammary eosinophils compared with strong expression in the blood while neutrophils were negative for these markers in both mammary washes and blood. Eosinophils in blood and mammary glands were negative for MHC class II, CD25 and CD4. Mast cells and lymphocyte aggregates were present in the tissue of glands chronically stimulated with parasite larvae while eosinophils were only present if the gland had been recently stimulated. These studies show that detailed in vivo analysis of leucocyte migration can be easily performed in the sheep mammary infusion model which allows non-invasive and repeated sampling of inflammatory cells before and after tissue migration.     

Cost-Effective Approach for Continuous Major Ion and Nutrient Concentration Estimation in a River

Major ion and nutrient concentration monitoring and estimation are important factors in management and interpretations on river health, particularly in the context of total maximum daily load limits. Spatial and temporal (daily, seasonally, yearly, etc.) variations commonly complicate investigations and can produce unrepresentative results, particularly in systems with large seasonal or daily variation in river parameters or concentrations as a result of physical loading or biogeochemical activity (e.g., photosynthesis and respiration). This study combines an observed relationship between electrical conductivity and major ions, including nitrate, and continuous colorimetric estimation of ammonium and phosphate to permit cost-effective real-time estimation of river concentrations for major ions and nutrients for surface water quality monitoring. Data collected from sites both up- and downstream of a major city were used to evaluate the method. Constant total dissolved solids (TDS) to electrical conductivity (EC) relationships were observed at both the upgradient (TDS = 696EC; r2 = 0.93) and downgradient (TDS = 684EC; r2 = 0.90) sites. The resulting predicted estimations of major ion and nutrient concentrations for each site had average errors of less than 5%. Combining this method with a modified continuous colorimetric method for ammonia and phosphate allows for the continuous estimation of major ion and nutrient concentrations in a river system.     

Ammonia-induced changes in pancreatic hormones and plasma amino acids in patients with liver cirrhosis

The contribution of hyperammonemia to plasma amino acid imbalance in patients with liver disease was assessed in 10 subjects with chronic hepatitis and in 17 advanced cirrhotics. Insulin, glucagon, and plasma amino acids were determined both in the basal state and 45 min after oral ammonium chloride, at doses used in the ammonia-tolerance test. In cirrhotics, ammonia increased to 3 times basal values, in association with a rise in insulin and, more marked, in glucagon. Aromatic amino acids and free tryptophan further increased, while a significant fall in branched-chain amino acids and glutamate was observed. The increase in ammonia levels strongly correlated with the increase in glucagon (r = 0.707). Two patients, with large esophageal varices, showed signs of disturbed consciousness, in association with a marked rise in ammonia and in the ration of free tryptophan to the sum of neutral amino acids. In patients with chronic hepatitis, whose ammonia levels rose slightly, minor variations in pancreatic glucoregulatory hormones and plasma amino acids were observed, as also happened in 10 healthy subjects following ammonium chloride ingestion. Our data fit with the hypothesis that the plasma amino acid imbalance of cirrhotics may be partly due to ammonia-induced changes in pancreatic hormones.     

Leaching of chrysocolla with Ammonia-Ammonium carbonate solutions

Chrysocolla was leached in solutions of ammonium hydroxide and ammonium carbonate as a function of the variables: temperature (25 to 55 ‡C), ammonia-ammonium ratio (0.0:1.0 to 1.0:0.0), total ammonia concentration (0.25 to 6.0 M), and particle size (100 to 400 mesh). A model of the leaching behavior was deduced based on: (1) the activation energy of 60.75 kJ/mole (14.51 kcal/mole) for 3 M total NH₃ which was dependent on both total ammonia concentration and temperature; (2) first-order dependence of rate on [(NH₄)₂CO₃]; (3) dependence of initial reaction rate on reciprocal of particle diameter; and (4) morphological evidence from SEM and ED AX measurements of diffusion and leaching occurring primarily in surface microcracks and not in the submicroscopic pores. In addition to the importance of diffusion through microcracks in rate control chemical reaction at active surface sites to produce the species, CuNH ₃ ^{2 +}, is also important. Only a fraction of the Cu atoms react that are exposed to lixiviant. Higher ammonia-ammonium ion concentrations, higher temperatures, or much longer times are required for more refractory Cu atoms to dissolve. Formerly Graduate Student, Department of Metallurgy and Metallurgical Engineering, University of Utah     

Effects of gastric mucosal blood flow (GMBF) on the role of adaptive cytoprotection of rat gastric mucosa

It has been reported that ingestion of an ammonium-containing diet produces hyperammonemia without encephalopathy, thus permitting the study of the specific effects of ammonia toxicity. The present study investigated the rat cerebral somatostatinergic system using this experimental model of hyperammonemia. Wistar rats were fed a high ammonia diet prepared by mixing a standard diet with ammonium acetate (20% w/w); in addition, 5 mM of ammonium acetate was added to their water supply. Control rats were fed with a standard diet. The animals were sacrificed at 3, 7 and 15 days of ammonia ingestion. Ammonia levels in blood had increased approximately 3-fold at 7 days of ammonia ingestion. These changes were associated with a significant decrease in the specific binding of somatostatin (SS) to putative receptors sites in the frontoparietal cortex and hippocampus at 7 and 15 days after starting the high ammonia diet. Scatchard analysis shows that the decrease in SS binding resulted from a decrease in the number of available SS receptors rather than a change in receptor affinity. No changes in the somatostatin-like immunoreactivity content (SSLI) were detected in either brain area at the three study times. These results suggest that hyperammonemia alone can affect the rat brain somatostatinergic system. However, the animal model of hyperammonemia used here is insufficient to produce encephalopathy despite the significant increase in serum ammonia.     

Solubility of ammonium paratungstate in aqueous diammonium phosphate and ammonia solution and its implications for a scheelite leaching process

Scheelite could be digested with an aqueous diammonium phosphate and ammonia solution in an autoclave. However, unexpected ammonium paratungstate (APT) crystallisation and tungstophosphoric acid formation occasionally occurred. In order to provide a theoretical basis for the scheelite leaching process, the solubility of APT in solution was determined by the isothermal solution equilibrium method. The effects of initial ammonia and diammonium phosphate concentration as well as temperature on APT solubility and tungstophosphoric acid formation were investigated. Results showed that APT solubility increased with increasing ammonia concentration, diammonium phosphate concentration and temperature, while tungstophosphoric acid concentration in the leaching solution increased with decreasing ammonia concentration and increasing diammonium phosphate concentration. In order to verify the effects of these factors on the leaching of scheelite, scheelite digestion experiments using diammonium phosphate and ammonia solution were also carried out.     

Mechanism of pyocyanin- and 1-hydroxyphenazine-induced lung neutrophilia in sheep airways

Pyocyanin (Pyo) and 1-hydroxyphenazine (1-HP) are extracellular products of Pseudomonas aeruginosa. To test whether these products were capable of producing an inflammatory response in the airways, combinations of Pyo and 1-HP at concentrations of 10(-4) and 10(-5) M were instilled into sheep airways, and indexes of inflammation were assessed by bronchoalveolar lavage (BAL) 24 h later. Challenge with the phenazines caused a significant dose-dependent increase in the number of cells and neutrophils recovered by BAL. Control challenges produced no such changes. The lung neutrophilia was accompanied by an increased concentration of albumin in BAL. The increases in BAL neutrophils and albumin could be blocked by treating the sheep with the 5-lipoxygenase inhibitor zileuton. Neither 1-HP nor Pyo was chemotactic to neutrophils when tested in vitro, but when alveolar macrophages (AM) were cultured in vitro in the presence of both Pyo and 1-HP (1 microM), the supernatants caused neutrophil chemotaxis. Analysis of AM culture supernatants incubated with the combination of pigments showed significant increases in leukotriene B4 and interleukin-8, and blocking these mediators separately or together reduced AM supernatant-induced neutrophil chemotaxis. We conclude that local instillation of Pyo and 1-HP can initiate an inflammatory response in the airways of sheep in vivo. This effect can be explained, in part, by the release of chemotactic factors produced by AM.     

Modeling Nitrogen Transport in Duckweed Pond for Secondary Treatment of Swine Wastewater

A mathematical model was developed to describe nitrogen transport in duckweed-covered static ponds for nutrient recovery from swine lagoon water. A finite difference technique was used to solve the partial differential equations describing the ammonia transport and concentration in the pond. The key parameters in the model include the diffusion coefficient of ammonium in the medium (D) and kinetic constant of nitrogen uptake by duckweed (k). Using one order of magnitude parameter variations, the simulations showed that the model was clearly much more sensitive to D than to k, indicating the process of nitrogen removal in a static pond by duckweed is diffusion limited. Laboratory testing was conducted with Spirodela punctata 7776, a duckweed strain, to calibrate the model. The calibration of the model with experimental data yielded a new ammonium transport coefficient (T) that is 85 times of D value. Model results showed good agreement with depth-wise experimental ammonium concentration and the model also demonstrates that intermittent mixing every 3 h can enhance ammonium uptake. Additionally, an apparent drop in pH near the duckweed mat at the surface was observed that may explain low rates of ammonia emission from duckweed ponds.     

氨法加压浸出钴铜氧化矿工艺

氨法浸出是基于目标金属与氨形成配合离子进入溶液,实现目标金属与部分杂质的分离,因此浸出过程具有选择性。对钴、铜与氨的配合机制及亚硫酸钠还原性能的影响因素进行了分析。结果表明:提高cNH3/cMe有利于形成稳定性高的钴、铜氨配合离子;降低cSO42-/cSO32-,提高体系pH可降低还原剂还原电位。实验过程采用加压氨浸工艺,在NH3-NH4+-H2O体系中浸出钴铜氧化矿中的钴和铜,研究了总氨浓度、氨铵比、液固比、浸出温度、还原剂用量对氧化矿中钴和铜浸出率的影响。结果表明,在总氨浓度7 mol.L-1、氨铵比2∶1、液固比6∶1、浸出温度100℃、还原剂亚硫酸钠用量为三价钴含量(摩尔比)4倍的最优条件下,钴浸出率可达到95.2%,铜浸出率可达到95.8%。浸出液后续处理工艺简单,氨及铵盐可实现闭路循环,对环境友好。     

氨浸液中游离氨对钼酸铵生产的影响

论述了钼酸铵中氨浸液的组成及主要组分的赋存状态，提出了氨浸液中游离氨浓度与钼酸铵生产的关系和作用机理，从理论和应用方面作了深入探讨。     

氨法浸出电镀废渣中镍铜的工艺

实验采用NH3-NH4+-H2O体系浸出电镀废渣中镍和铜,通过正交试验研究了总氨浓度、氨铵比、液固比、温度、浸出时间对浸出率的影响.结果表明,在总氨浓度为6 mol/L、氨铵比为1∶1、液固比为8∶1、浸出温度80℃、浸出时间3h的最优条件下,镍的浸出率可达到82％,铜的浸出率可达到95％.     

Effect of Fasciola hepatica excretory-secretory products on the metabolic burst of sheep and human neutrophils

F. hepatica excretory-secretory (ES) products were found to inhibit superoxide output in phorbol myristate acetate (PMA)-stimulated sheep and human neutrophils as measured by spectrophotometry. Luminol-enhanced chemiluminescence by PMA-stimulated neutrophils from both species was again inhibited. However, nitric oxide output by PMA-stimulated human neutrophils was significantly increased in the presence of ES products, whilst in sheep it was inhibited. No major effects were noted using resting neutrophils. The results are discussed in relation to the evolution of parasite defence mechanisms.