Extraction of DNA from amniotic fluid cells for the early prenatal diagnosis of genetic disease

Ten-ml samples of amniotic fluid were taken from pregnancies being terminated at 8–14 weeks' gestation. DNA was extracted from the amniotic cells by sequential centrifugation and analysed using the polymerase chain reaction (PCR). Fifteen samples were analysed for evidence of maternal contamination using Mfd5 oligo-nucleotide primers for repeat polymorphisms. Ten amniotic fluid samples were tested for the Delta-F₅₀₈ deletion characteristic of cystic fibrosis to demonstrate a diagnostic application for the technique. In each case, DNA extracted from fetal tissue from the same pregnancy was included in the controls. In 14 of the 15 cases tested with the Mfd5 primers, both the amniotic fluid DNA and the fetal DNA showed no evidence of contaminating DNA. In one case, neither the amniotic fluid cells nor the fetal cells yielded results. In nine of the ten cases tested with the Delta-F₅₀₈ primers, the amniotic fluid cell DNA provided accurate information about the genetic status of the fetus; in the tenth, the fetal DNA failed to amplify. The results indicate that adequate DNA can be extracted from amniotic fluid from 8 weeks' gestation onward and these samples are suitable for prenatal diagnosis using PCR.     

The effect of metapleural gland secretion on the growth of a mutualistic bacterium on the cuticle of leaf-cutting ants

In Acromyrmex octospinosus leaf-cutting ants the metapleural glands produce an array of antibiotic compounds that serve as a general defence against unwanted microbes on the cuticle. Leaf-cutting ants also grow mutualistic Pseudonocardiaceae bacteria on their cuticle that produce antibiotics controlling a microfungal parasite of their fungus gardens. Interaction between this bacterium and gland secretion therefore seems unavoidable. We document the typical development of bacterial growth on the cuticle of young major workers, show that growth starts a few days after eclosion, and that the maximal cover is reached after 2–3 weeks and gradually declines when workers mature. Experimental closure of the metapleural glands had no effect on the initial exponential growth phase of the bacterium, but significantly reduced the cover during the decline phase. The age-dependent abundance of the bacterium and its partial dependence on metapleural gland secretion support the hypothesis that the abundance of this mutualist is actively regulated.     

The influence of source and timing of nitrogen fertilizers on yield and nitrogen use efficiency of four sweet potato cultivars

Nitrogen (N) availability is a very important determinant of crop yield. It is also one of the most expensive inputs in crop production. Thus, the efficient management of N by farmers with limited resource is a very important part of successful soil and crop management system. A field study was conducted on a Norfolk sandy loam soil at the George Washington Carver Agricultural Experiment Station, Tuskegee, AL, to evaluate the influence of fertilizer source and timing on the yield of four sweet potato [Ipomoea batatas (L.)] cultivars. Two N sources (urea, CO(NH₂)₂ and ammonium nitrate, NH₄NO₃), four sweet potato cultivars (‘Georgia Jet’, GA-Jet; ‘TU-82-155’, TU-155; ‘TU-1892’; and ‘Rojo Blanco’, Rojo-Bl) and three times (zero, single, and split applications) were used in a factorial designed experiment. Plants were harvested after 80 and 120 days and total and marketable yields determined. Nitrogen recovery efficiency, yield efficiency and physiological efficiency were also determined. Total and marketable yields of early maturing cultivars were significantly higher than of late maturing cultivars (P<0.05). The single application of N resulted in significantly higher yield in storage roots than split application (P<0.05). Physiological efficiency values were highly correlated with total marketable yields. Recovery rates increased with time. Late maturing cultivars tended to have higher N recovery and physiological efficiency than early maturing cultivars. These results suggest that cultivar maturity group should play an important part in N fertilization recommendations for sweet potato.     

Profitability of intensification technologies among smallholder maize farmers in the forest-savanna transition zone of Nigeria

Sustaining the productive capacity of the land resource under the existing land-use system and increasing population pressure requires the use of appropriate technologies that will enhance farm productivity and income levels.This paper examines the effect on farm profitability of smallholder maize farmers’ use of intensification technologies in tropical agriculture. The survey was conducted in southwest Nigeria. A multi-stage sampling technique was used to select 300 respondents from two major agroclimatic zones in Osun State. Primary data were collected using a structured questionnaire and complemented with focus group discussions (FGDs), while the State’s Ministry of Agriculture provided secondary data.Pressure on land was found high because farmland was continuously cropped for an average of 10 years with a mean fallow period of 2 years only. Technologies used to intensify agriculture included inorganic fertilizer, organic manure, alley cropping, and tree planting.Results from costs and returns analysis showed that average net returns to inorganic fertilizer use ranked highest in the two zones, while tree planting ranked lowest. There was a significant ecoregional difference on the average net returns/year earned by users of mineral fertilizer. While organic manure use earned higher net returns than alley cropping practice in the rain forest, the reverse was the case for the derived savanna. FGDs revealed that respondents’ preferred to use inorganic fertilizer despite its inadequate supply because, apart from enhancing higher returns and net gains, it also improved output level and provided an opportunity for the continuous use of the scarce land.These results imply that economic returns play a critical role in farmers’ use of intensification technologies and consequently affect their resource management decisions. Policy measures aimed at promoting research on appropriate technologies that are profitable on farmers’ fields will be more effective in enhancing farmers’ use of intensification technologies.     

Groundwater pollution due to discharge of tannery effluents in Upper Palar basin, Tamilnadu, India: an assessment through mass transport modeling

The groundwater regime in Upper Palar basin, Tamilnadu has been highly contaminated in several locations due to discharge of effluents from a large number of tanneries. At some places total dissolved solids (TDS) concentration in groundwater was found as high as 8000 mg/l. Transmissivity and storativity of the regional aquifer were estimated at a few locations. The porosity and dispersivity values were not determined in the field. These parameters were assumed based on data available for similar geological formations elsewhere. The aquifer conceptualization thus arrived at formed the basis of a numerical groundwater flow model which was constructed using the finite difference method. The flow model was calibrated for steady state and then for transient condition for the period of 1984-92. The computed heads and calibrated parameters of the flow model were used to compute groundwater velocities. The migration of contaminants for a 20 year period was computed using the hydraulic heads and effective porosity value in a pathline model using FLOWPATH software. Mass transport model was constructed using Method of Characteristics (MOC) computer code in a separate model. The seepage rate of effluent is assumed at a rate of 30% of that discharged on the surface. The mass concentration of solute in the effluent reaching the water table was assumed as 40%, the same as in the surface effluent. The mass transport model was calibrated for a 20 year period. Prediction of contaminant migration from different clusters in the basin was analyzed. The prediction results indicated elevated TDS concentration of more than 4000 mg/l from most clusters. Also the area of the contaminated zone is likely to double in 20 years from contaminated zone of 1992.     

Breccias of the Adnet Formation: indicators of a Mid-Liassic tectonic event in the Northern Calcareous Alps (Salzburg/Austria)

Stratigraphy, lithology and depositional structures of Liassic red limestone-breccias of the Adnet Formation, including the 'Adnet Scheck', were studied at several outcrops of the Northern Calcareous Alps (NCA) south-east of Salzburg. A four-fold lithostratigraphic division is proposed for the Adnet Formation of the Osterhorn Mountains: the hemipelagic Schmiedwirt (Sinemurian) and Kehlbach (Carixian) members are separated from the pelagic Saubach Member (Toarcian) by a layer of amalgamated breccias (Scheck Member, probably Domerian to early Toarcian). Several other breccia beds occur locally from the base of the Kehlbach Member up to the lower Saubach Member. Although the sediments overlying the Scheck Member breccias are of coeval age, the ages of the underlying strata are very different. This can be explained by submarine Liassic erosion during a period of resedimentation from the middle Carixian until the early Toarcian. At least 10–15?m of partly lithified sediments were eroded by gravity flows. The entire Kehlbach Member and up to two-thirds of the Schmiedwirt Member were removed at Adnet. The breccias originated from submarine debris flows. Repeated flows over a long period and the depositional setting exclude a triggering by sea-level fluctuations. Most probably they arose from tectonically triggered slumps and slides of superficial sediments. The 'Scheck' was initiated on the steep upper slope of the drowned Triassic Adnet reef and flowed to the north-east.The Pliensbachian to early Toarcian period of tectonic activity indicated by the breccias was the most important during the Liassic in the Osterhorn Mountains and other parts of the NCA. From the large-scale regional distribution of the breccias and in accord with published data, a roughly north-east trending strike-slip fault zone is proposed, crossing the NCA south of the Osterhorn block, with a peaking activity during the Pliensbachian to early Toarcian as the cause of the tectonic movements.     

Latest precambrian (Cadomian) zircon ages, Nd isotopic systematics and P-T evolution of granitoid orthogneisses of the Erzgebirge, Saxony and Czech Republic

Single zircons from two orthogneiss complexes, the Grey Gneiss and Red Gneiss, the lowermost tectonic units in the Erzgebirge, were dated. The grey Freiberg Gneiss is of igneous origin and has a ²⁰⁷Pb/²⁰⁶Pb emplacement age of 550±7 Ma. A quartz monzonite from Lauenstein contains idiomorphic zircons with a mean ²⁰⁷Pb/²⁰⁶Pb age of 555±7 Ma as well as xenocrysts ranging in age between 850 and 1910 Ma. Red gneisses from the central Erzgebirge contain complex zircon populations, including numerous xenocrysts up to 2464 Ma in age. The youngest, idiomorphic, zircons in all samples yielded uniform ²⁰⁷Pb/²⁰⁶Pb ages between 550±9 and 554±10 Ma. Nd isotopic data support the interpretation of crustal anatexis for the origin of both units. _Nd(t) values for the grey gneisses are –7.5 and –6.0 respectively, (mean crustal residence ages of 1.7–1.8 Ga). The red gneisses have a wider range in _Nd(t) values from –7.7 to –2.8 (T _DM ages of 1.4–1.8 Ga). The zircon ages document a distinct late Proterozoic phase of granitoid magmatism, similar in age to granitoids in the Lusatian block farther north-east. However, Palaeozoic deformation as well as medium pressure metamorphism ( 8 kbar/600–650° C) are identical in both gneiss units and distinguish these rocks from the Lusatian granitoids. The grey and red gneisses were overthrust by units with abundant high-pressure relicts and a contrasting P-T evolution. Zircon xenocryst and Nd model ages in the range 1000–1700 Ma are similar to those in granitoid rocks of Lusatia and the West-Sudetes, and document a pre-Cadomian basement in parts of east-central Europe that, chronologically, has similarities with the Sveconorwegian domain in the Baltic Shield.     

Chorionic villus sampling: Analysis of fetal losses to delivery,placental pathology,and cervical microbiology

A population of 1639 patients were seen for chorionic villus sampling (CVS). Embryonic death was identified at ultrasound in 5.3 per cent of patients. The number of patients undergoing CVS was 1551, with 1416 transcervical procedures and 135 transabdominal procedures. The most common indication for CVS was advanced maternal age. Spontaneous pregnancy losses identified by increased risk of pregnancy loss with increasing aspiration attempts. The total fetal loss for this population was 5.4 per cent with the pregnancy loss estimated due to procedure being 1.2 per cent. Analysis of placentae from patients having CVS and amniocen-tesis showed no differences. Microbiological assessment prior to CVS was similar to previous publications.