Cell death localization in situ in laboratory reared honey bee (Apis mellifera L.) larvae treated with pesticides

In this study, cell death detected by DNA fragmentation labeling and phosphatidylserine (PS) localization was investigated in the honey bee (Apis mellifera L.) midgut, salivary glands and ovaries after treating larvae with different pesticides offered via an artificial diet. To do this, honey bee larvae reared in an incubator were exposed to one of nine pesticides: chlorpyrifos, imidacloprid, amitraz, fluvalinate, coumaphos, myclobutanil, chlorothalonil, glyphosate and simazine. Following this, larvae were fixed and prepared for immunohistologically detected cellular death using two TUNEL techniques for DNA fragmentation labeling and Annexin V to detect the localization of exposed PS specific in situ binding to apoptotic cells. Untreated larvae experienced ∼10% midgut apoptotic cell death under controlled conditions. All applied pesticides triggered an increase in apoptosis in treated compared to untreated larvae. The level of cell death in the midgut of simazine-treated larvae was highest at 77% mortality and statistically similar to the level of cell death for chlorpyrifos (65%), imidacloprid (61%), myclobutanil (69%), and glyphosate (69%) treated larvae. Larvae exposed to fluvalinate had the lowest midgut columnar apoptotic cell death (30%) of any pesticide-treated larvae. Indications of elevated apoptotic cell death in salivary glands and ovaries after pesticide application were detected. Annexin V localization, indicative of apoptotic cell deletion, had an extensive distribution in the midgut, salivary glands and ovaries of pesticide-treated larvae. The data suggest that the tested pesticides induced apoptosis in tissues of honey bee larvae at the tested concentrations. Cell death localization as a tool for a monitoring the subclinical and sub-lethal effects of external influences on honey bee larval tissues is discussed.     

Follow-up of mercury levels in fish,human hair and urine in the Madeira and Tapajós basins,Amazon, Brazil

Informal economy of gold mining has contaminated some important river basins in Amazon. Follow-up studies on critical compartments showed some areas with high Hg levels in fish as well as in human hair samples. Average Hg in piscivorous fish in the Madeira river itself was 846 ppb (N=284) with a maximum of 3921 ppb. Mercury in fish from non polluted areas in this basin shows high variability, even for single species. A seasonal variation in Hg content was observed, with higher values at the end of the dry season. In the upper Tapajós basin comparable values were found for fish but with a definite decreasing trend downstream. Average value for piscivorous fish in the whole Tapajós basin is 482 ppb (N=122) with a maximum value of 3770 ppb. Hair Hg was higher in fishing villages in the Tapajós (average: 17 ppm; with N=432 and maximum value of 176 ppm) than in the Madeira (average: 9 ppm; N=169; maximum 71 ppm), and data from some areas of the Tapajós suggest a decrease with time. Mercury was much higher in urine of goldshop workers in Santarém (low Tapajós) than in Alta Floresta (high Tapajbs) and show a decreasing trend in both cases, probably related to the significant decline in gold mining activities during the study period (1986–1994).     

Significant increases in pulping efficiency in C4H-F5H-transformed poplars: improved chemical savings and reduced environmental toxins

The gene encoding ferulate 5-hydroxylase (F5H) was overexpressed in poplar (Populus tremula x Populus alba) using the cinnamate-4-hydroxylase (C4H) promoter to drive expression specifically in cells involved in the lignin biosynthetic pathway and was shown to significantly alter the mole percentage of syringyl subunits in the lignin, as determined by thioacidolysis. Analysis of poplar transformed with a C4H-F5H construct demonstrated significant increases in chemical (kraft) pulping efficiency from greenhouse-grown trees. Compared to wild-type wood, decreases of 23 kappa units and increases of >20 ISO brightness units were observed in trees exhibiting high syringyl monomer concentrations. These changes were associated with no significant modification in total lignin content and no observed phenotypic differences. C4H-F5H-transformed trees could increase pulp throughputs at mills by >60% while concurrently decreasing chemicals employed during processing (chemical pulping and bleaching) and, consequently, the amount of deleterious byproducts released into the environment.     

Re-analysis of species associational data using bootstrap significance tests

Re-analysis with the bootstrap technique of species associational data obtained in 1973 at a waste discharge site showed that stations previously accepted as apparently similar could actually be significantly different. Two station pairs with almost identical similarities (Bray-Curtis Coefficients of 0.85 and 0.84) were found to be significantly different (P 0.040) in the first case, but not significantly different in the second (P 0.78). The significantly different pair reflected differences that can arise between sampling stations impacted by light deposits of mine tailings. The other clustered pair (not significantly different) comprised two depauperate stations that had received heavy tailings deposition. A similarity of 0.67 between samples obtained using two types of samplers (Van Veen and Ponar) at one station showed a significant difference (P 0.0002). This supported previous suspicion that the samplers were collecting differently, although this had not been demonstrated by the cluster analysis used in 1973. However the use of either sampler's data in the cluster analysis had only trivial effect on similarity levels, and no effect on the significance tests.     

Direct drilling,shallow tine-cultivation and ploughing on a silt loam soil, 1974–1980

Results are given for an experiment in which direct drilling, shallow tine-cultivation and ploughing were compared on a silt loam soil over 7 years. A rotation of cereal and oilseed rape crops was established to minimise a possible interaction between disease, particularly take-all (Gaeumannomyces graminis), and cultivation treatment. Over the six harvest years shallow tillage produced on average a 6 and 9% heavier yield than ploughing and direct drilling, respectively. The importance of surface soil conditions at the time of seedling establishment was clearly evident. In years when the passage of the direct drill caused little soil disturbance and seeds were placed in a smeared slot, and particularly when crop residues were pressed into the slot, plant populations were diminished and so were yields. The degree of soil shattering by the passage of the drill and the consequent friability of the tilth varied between the extremes of water content of the top soil (0–5 cm). Direct drilling produced larger yields relative to ploughing after the first three seasons and this may reflect the increased organic matter content and stability of the soil aggregates in the surface layer (0–2.5 cm) which have already been reported for this soil. These changes may have facilitated the greater friability of the soil and the creation of tilth by the passage of the drill which ensured uniform germination and rapid establishment of the seedlings.The site was characterised by variations in the depth of topsoil over gravel (< 50 to > 100 cm), and the deepest soil gave the heaviest yield. This effect was relatively greater in dry seasons but it never interacted with the effect of tillage method.     

Exploring the spatial relations between cereal yield and soil chemical properties and the implications for sampling

Abstract. In general, agricultural management has focused on differences between fields or on the gross differences within them. Recent developments in agricultural technology, yield mapping, Global Positioning Systems and variable rate applications, have made it possible to consider managing the considerable variation in soil and other properties within fields. This system is known as precision agriculture. More precise management of fields depends on a better understanding of the factors that affect crop input decisions. This paper examines the spatial variation in crop yield, soil nutrient status and soil pH within two agricultural fields using geostatistics. The observed properties vary considerably within each field. The relation between yield and the measured soil properties appears to be weak in general. However, the range of spatial correlation for yield, shown by the variogram, is similar to that of the soil chemical properties. In addition the latter changed little over two years. This suggests that information on the scale of variation of soil chemical properties can be derived from yield maps, which can also be used as a guide to a suitable sampling interval for soil properties.     

Evaluation of First-Feeding Regimens for Larval Nassau Grouper Epinephelus straitus and Preliminary, Pilot-Scale Culture through Metamorphosis

Two 10-day hatchery experiments were conducted to evaluate s-type (Hawaiian strain) and ss-type (Thailand strain) rotifers Brachionus plicatilis and cryogenically preserved oyster Crassostrea gigas trochophores as first feeds for larval Nassau grouper Epinephelus striatus. Newly hatched grouper larvae were reared at densities of 11.2–20.8/L in 500-L tanks at 36–38 ppt salinity, 25–26 C, and under a 11-h light: 13-h dark photoperiod. Beginning on day 2 posthatching (d2ph), prey were maintained at a density of 20 individuals/mL, while phytoplankton (Nanochloropsis oculata) was maintained at 500 × 10³ cells/mL. In experiment 1, survival and growth were higher (P < 0.05) for fish fed small s-type rotifers (mean lorica length = 117 μm; fish survival = 7.96%) selected by sieving than for fish fed non-selected rotifers (mean lorica length = 161 μm; fish survival = 2.13%). These results demonstrated the advantage of small prey size and suggested that super-small (ss-type) rotifer strains would be beneficial. In experiment 2, three feeding regimens were compared: 1) ss-type rotifers (mean lorica length = 147 μm); 2) oyster trochophores (mean diameter = 50 μm) gradually replaced by ss-type rotifers from d5ph; and 3) a mixed-prey teatment of 50% oyster trochophores and 50% ss-type rotifers. Survival was higher (P < 0.05) for larvae fed mixed prey (15.6%) than for those fed rotifers (9.73%) or trochophores and rotifers in sequence (2.55%), which also showed the slowest growth. Oyster trochophores, although inadequate when used exclusively, enhanced survival when used in combination with rotifers, possibly by improving size selectivity and dietary quality. In a pilot-scale trial, larvae were cultured through metamorphosis in two 33.8-m³ outdoor tanks. Fertilized eggs were stocked at a density of 10 eggs/L and larvae were fed ss-type rotifers from d2ph-d20ph, newly hatched Artemia from d15ph-d18ph, 1-d-old Artemia nauplii from d18ph-d62ph. Survival on d62ph was 1.17%, with a total of 5,651 post-metamorphic juveniles produced.     

Strain variation in Dermatophilus congolensis demonstrated by cross-protection studies

Cross-protection studies were conducted with vaccines prepared from two isolates of Dermatophilus congolensis (designated strain 1 and strain 2). The vaccines were prepared as either heat-inactivated, washed, formalized filamentous phase bacterium, mixed with alum as an adjuvant, and inoculated intramuscularly (type A vaccine) or sedimented live filaments inoculated intradermally (type B vaccine). The vaccinated sheep were challenged with D. congolensis zoospores of one or other strain. Challenge sites were observed for the presence and severity of lesions. Serum antibody levels to D. congolensis were monitored after vaccination and challenge. Type A and B vaccines from both strains produced some reduction in the severity of lesions when sheep were challenged with strain 1 but not with strain 2. Unvaccinated control sheep developed more severe and persistent lesions when challenged with strain 2 than controls challenged with strain 1. Serum antibody levels to the type B vaccine prepared from strain 1 were significantly higher (P less than 0.05) than antibody levels to type B vaccine from strain 2. These findings showed there was significant variation in virulence and antigenicity between these two isolates of D. congolensis.     

Experimental reproduction of ovine vulvitis with bacteria of the haemophilus/histophilus group

Ovine vulvitis was experimentally reproduced by intravaginal inoculation of a mixture of bacterial strains of the histophilus/haemophilus group isolated from field cases. Grossly, the experimental vulvitis was identical to the field condition, and bacteria indistinguishable from the inoculated strains were reisolated.     

Clinical and immunologic response of cattle to administration of a vaccine containing modified-live bovine respiratory syncytial virus

Healthy yearling beef and dairy cattle were inoculated with a vaccine containing modified-live bovine respiratory syncytial virus (ML-BRSV), and sequential changes in clinical signs of disease, blood leukocyte subsets, BRSV-specific antibody titer, and in vitro lymphocyte blastogenic responses were monitored. Vaccination with ML-BRSV did not cause pyrexia, local or systemic hypersensitivity reaction, or respiratory tract disease. Episodes of leukopenia, abnormalities in lymphocyte subsets, or depression of phytomitogen-induced blastogenic responses were not observed subsequent to vaccination. Exposure to ML-BRSV resulted in at least a 16-fold increase in serum neutralizing antibody titer, with no increase seen in nonvaccinated contact controls. Significant BRSV-specific lymphocyte blastogenic responses were not detected, using one dose of several BRSV antigen preparations in a whole blood culturing system.