Synthesis of immunoglobulin by normal and antigenically stimulated fetal sheep.

The rate of appearance, quantity and immunochemical character of serum immunoglobulins which appear normally during the development of fetal sheep and following the injection of antigens at different stages of gestation have been studied. After 70 days' gestation a percentage of normal fetal sheep synthesise IgM. Although the concentration of IgM in the circulation of these animals was very low, the precentage with IgM increased with fetal age. A few late term fetuses were detected which also had IgG1 in their circulation, although none were detected with IgG2 or IgA. Fetuses injected with antigens before 71 days' gestation only synthesised IgM, while fetuses injected after 79 days' gestation synthesised both IgM and IgG1. Neither IgG2 nor IgA were detected by single-radial immunodiffusion analyses during the first 14 days of primary immune responses to a variety of antigens, although trace amounts of IgG2 were detected late in responses occurring in older fetuses. The immunoglobulins synthesised by antigenically stimulated fetal sheep appeared identical to adult sheep 19S IgM, 7S IgG1 and 7S IgG2 respectively, when analysed by immunoelectrophoresis, isoelectric focusing and G200 Sephadex column chromatography.     

Aluminum-induced decrease in CO2 assimilation in citrus seedlings is unaccompanied by decreased activities of key enzymes involved in CO2 assimilation

'Cleopatra' tangerine (Citrus reshni Hort. ex Tanaka) seedlings were irrigated daily for 8 weeks with 1/4 strength Hoagland's nutrient solution containing 0 (control) or 2 mM aluminum (Al). Leaves from Al-treated plants had decreased CO2 assimilation and stomatal conductance, but increased intercellular CO2 concentrations compared with control leaves. On a leaf area basis, 2 mM Al increased activities of key enzymes in the Calvin cycle, including ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), stromal fructose-1,6-bisphosphatase (FBPase), and a key enzyme in starch synthesis, ADP-glucose pyrophosphorylase (AGPase), compared with control leaves. Aluminum had no effect on cytosolic FBPase activity, but it decreased sucrose phosphate synthase (SPS) activity. Aluminum had no effect on area-based concentrations of carbohydrates, glucose-6-phosphate (G6P) and fructose 6-phosphate (F6P) or the G6P:F6P ratio, but it decreased the area-based concentration of 3-phosphoglycerate (PGA). Photochemical quenching coefficient (qP) and electron transport rate through PSII were greatly reduced by Al. Non-photochemical quenching coefficient (NPQ) was less affected by Al than qP and electron transport rate through PSII. We conclude that the reduced rate of CO2 assimilation in Al-treated leaves was probably caused by a combination of factors such as reduced electron transport rate through PSII, increased closure of PSII reaction centers and increased photorespiration.     

Effects of Nitrogen Rate and Harvest Moisture Content on Physicochemical Properties and Milling Yields of Rice

Field studies were conducted from 2011 to 2013 near Stuttgart, Arkansas. The impacts of nitrogen rate (0, 45, 90, 135, and 180 kg of N/ha) and harvest moisture content (HMC) (23, 19, and 15%, wet basis) on physicochemical properties and milling yields were determined. Trends were similar for the cultivars evaluated: Cheniere, CL XL745, and Wells. Milled rice yields were only minimally impacted by either N rate or HMC level. Increasing N rate reduced kernel length and thickness of brown rice, chalkiness of brown rice and head rice, and viscosities of head rice flour, and it increased brown rice and head rice crude protein content and head rice yield (HRY). In terms of milling yields and head rice functionality, these data suggest that N rates as low as 90 kg of N/ha could be utilized, should production recommendations be changed. Significant interactions between N rate and HMC level were infrequent and were associated with the 0 kg of N/ha rate, unrealistic for rice production. Decreasing HMC from 23 to 19% reduced kernel length and thickness and increased crude protein content and chalkiness; further decreasing HMC to 15% also increased kernel fissuring and decreased HRY.     

Detection of castor contamination by real-time polymerase chain reaction

Due to the potential for intentional contamination of food with crude preparations containing ricin, a real-time PCR method was developed for the detection of castor plant material in ground beef. One primer pair was identified and confirmed to be castor-specific and efficient for amplification of ricin in DNA extracts from castor or beef matrices. Of three different DNA extraction protocols compared, the hexadecyltrimethylammonium bromide (CTAB) method yielded the highest quality of DNA for QPCR assay. The detection limit for castor contamination in ground beef samples was <0.001% (<10 microg of castor acetone powder per gram of beef, corresponding to 0.5 microg of ricin), indicating excellent sensitivity for the assay, well below the threshold for oral toxicity.     

Insoluble fraction of buckwheat (Fagopyrum esculentum Moench) protein possessing cholesterol-binding properties that reduce micelle cholesterol solubility and uptake by Caco-2 cells

Buckwheat (Fagopyrum esculentum Moench) protein (BWP) exhibits hypocholesterolemic activity in several animal models by increasing fecal excretion of neutral and acidic sterols. In the current study, the ability of BWP to disrupt micelle cholesterol solubility by sequestration of cholesterol was investigated. When BWP (0.2%) was incubated with cholesterol and micelle lipid components prior to micelle formation, cholesterol solubility was reduced 40%. In contrast, cholesterol solubility was not decreased when BWP (0.2%) was incubated after micelle formation and incorporation of soluble cholesterol. Buckwheat flour, from which BWP was derived, had no significant effect on cholesterol solubility. Cholesterol uptake in Caco-2 cells from micelles made in the presence of BWP (0.2%) was reduced by 47, 36, 35, and 33% when compared with buckwheat flour, bovine serum albumin, casein, and gelatin, respectively. Reduction in cholesterol uptake in Caco-2 cells was dose-dependent, with maximum reductions at 0.1-0.4% BWP. In cholesterol-binding experiments, 83% of the cholesterol was associated with an insoluble BWP fraction, indicating strong cholesterol-binding capacity that disrupts solubility and uptake by Caco-2 cells.     

T lymphocyte responses during early enteric Mycobacterium avium subspecies paratuberculosis infection in cattle

Johne's disease (JD) is a costly intestinal disease of ruminants caused by Mycobacterium avium subspecies paratuberculosis (Map), which is transmitted to perinatal calves by the fecal-oral route. Disease control efforts focus on identification and culling of infected cattle from herds; therefore failure to identify animals early is a major obstacle to reducing transmission. Development of host immunity during early JD remain incompletely characterized so detecting subclinical JD using immunologic techniques is a substantial challenge in the field. Development of a test with high sensitivity and specificity is a major research goal with significant implications for the cattle industry. The objectives of this study were to compare early Map-specific T lymphocyte responses in naive, experimentally Map infected and Map vaccinated calves using a subcutaneous matrigel biopolymer-based assay. We examined the phenotype of recruited lymphocytes and local interferon gamma (IFNγ) production within subcutaneously placed matrigel containing Map antigen 30 days after experimentally induced intestinal Map infection or Map vaccination. We show that IFNγ-secreting CD4+ T cells are recruited to matrigel sites in vaccinated but not infected or naïve calves. γδ T cells recruited to matrigel sites of Map-infected calves were mostly WC1-, while γδ T cells recruited to matrigel sites of Map-vaccinated calves were predominantly WC1+. IFNγ at matrigel sites was a discriminating factor between infected calves, naïve calves and vaccinated calves. These data contribute to our understanding of early anti-Map immunity, and may be useful for detecting early intestinal Map infections in calves or for enhancing our ability to discriminate between Map-infected and Map-vaccinated calves.     

An investigation of avoidance by Antarctic krill of RRS James Clark Ross using the Autosub-2 autonomous underwater vehicle

The autonomous underwater vehicle (AUV) Autosub-2 was deployed on eight missions ahead of RRS James Clark Ross in the northern Weddell Sea and in the Bransfield Strait, Southern Ocean, to assess avoidance of the research vessel by Antarctic krill Euphausia superba. The AUV was equipped with the same type of scientific echosounder as the research vessel (Simrad EK500 operating at 38 and 120 kHz) and measured the density of krill along transect acoustically (g m⁻² wet mass) prior to the ship’s arrival. We hypothesised that if krill avoided the ship, perhaps in response to radiated noise, then the ship should detect less krill than the AUV which is known to have much lower noise levels than the ship. We were unable to detect any significant difference between the density of krill detected by the ship or the AUV, either at the transect level or at finer scales within transects. We conclude, therefore, that avoidance by krill of RRS James Clark Ross will not significantly bias acoustic estimates of krill abundance by this vessel.     

Translocation of cave fish (Poecilia mexicana) within and between natural habitats along a toxicity gradient

During ecological speciation, diverging taxa have the potential to remain in close spatial proximity (i.e., sympatry or micro‐allopatry) theoretically allowing for continued contact and gene flow. In a system where incipient speciation of populations of the Atlantic molly (Poecilia mexicana) appears to be driven by abiotic factors, we investigated whether one of these factors, hydrogen sulphide (H₂S) toxicity, also constitutes an effective barrier to slow migration within and between habitats. We addressed this experimentally by translocating individuals from high toxicity to lower toxicity within a toxic cave and by translocating individuals from the toxic cave to a nontoxic surface habitat. Using a stepwise‐backwards Cox regression, we found that overall mortality was low, but statistically significant mortality occurred when individuals were transferred from higher toxicity to lower toxicity. In addition, only males were negatively affected by being transferred from low levels of toxicity to nontoxic, surface waters. This indicates that in addition to abiotic factors, other mechanisms, like predation and sexual selection, must be important in maintaining population separation.