Fate of benzoxazinone allelochemicals in soil after incorporation of wheat and rye sprouts

Growing cereals (especially rye), which are incorporated into the soil to increase soil fertility or organic matter content, is a common practice in crop rotation. The additional sanitizing effect of this incorporation has often been appreciated and is said to be due to leaching of benzoxazinones and subsequent formation of benzoxazolinones. In this study wheat (Stakado) and rye (Hacada) sprouts were incorporated into soil in amounts that simulated agricultural practice. By extraction and subsequent LC-MS analysis the disappearance and appearance of benzoxazinones, benzoxazolinones, and phenoxazinones in soil were followed. In the wheat experiments 6-methoxybenzoxazolin-2-one (MBOA) was detected as the main compound. 2-Hydroxy-7-methoxy-1,4-benzoxazin-3-one (HMBOA) and 2-hydroxy-1,4-benzoxazin-3-one (HBOA) were detected as well. No phenoxazinones were detected. For the rye experiment the picture was more complex. In the first 2 days of incubation MBOA and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA) were detected as the main allelochemicals along with HBOA, HMBOA, and benzoxazolin-2-one (BOA), in decreasing order. Later in the incubation period some 2-amino-3H-phenoxazin-3-one (APO) was detected and the amount of HBOA increased considerably and decreased again. The profiling of the benzoxazinone metabolites and their derivates in soil was dynamic and time-dependent. The highest concentrations of most of the compounds were seen at day 1 after incorporation. A maximum concentration was reached at day 4 for a few of the compounds. This study is the first of its kind that shows the dynamic pattern of biologically active benzoxazinone derivates in soil after incorporation of wheat and rye sprouts. Methods for organic synthesis of HBOA and HMBOA were developed as part of the study.     

Transformation of benzoxazinones and derivatives and microbial activity in the test environment of soil ecotoxicological tests on Poecilus cupreus and Folsomia candida

Benzoxazinones, such as 2,4-dihydroxy-7-methoxy-1,4-benzoxazin-3-one (DIMBOA) and 2,4-dihydroxy-1,4-benzoxazin-3-one (DIBOA), and benzoxazolinones, such as 6-methoxy-2-benzoxazolinone (MBOA) and 2-benzoxazolinone (BOA), are biologically active secondary metabolites found in cereals. Because these compounds could be exploited as part of a strategy for reducing the use of synthetic pesticides, ecotoxicological tests were performed recently. In this paper, the transformation of the compounds in the test environment of the ecotoxicological tests was studied. DIMBOA was degraded and partly transformed to MBOA during the period of ecotoxicological testing of the compounds. During testing of MBOA on Poecilus cupreus test media the analysis showed that at the initial concentrations of 2 and 10 mg kg(-1) no MBOA was left after 45 days of testing, but the metabolite 2-amino-phenoxazin-3-one (AMPO) was formed. During testing of BOA on both Folsomia candida and Poecilus cupreus the more biologically active compound 2-amino-phenoxazin-3-one (APO) was formed. Thus, the ecotoxicological test results on MBOA and BOA were partly due to the microbial transformation of the compounds during the time of testing.     

Chemical ecology in wheat plant-pest interactions. how the use of modern techniques and a multidisciplinary approach can throw new light on a well-known phenomenon: allelopathy

A new holistic approach on research into allelopathy was launched in the FATEALLCHEM project. The project was financed by the European Commission in the 5th Framework Programme and involved agronomists, biologists, analytical chemists, organic chemists, environmental chemists, ecotoxicologists, and modelers. Benzoxazinones from wheat (Triticum aestivum L.) were the main group of compounds studied in the project. The project showed that future assessments of an extensive use of allelopathic crops must include the development of validated analytical methods, considerations of relevant concentrations, studies on soil transformation, ecotoxicological studies on individual compounds and mixtures, evaluation on human and mammal toxicity, and joint effect studies on weeds, insects, and pathogens. The project results clearly showed the relevance of optimizing the exploitation of cereal benzoxazinones. Crop rotation is a very traditional practice, which was given less importance for decades but is now regaining its importance in agricultural practice as a means of controlling weed seed banks and soilborne diseases and pests. When using cereals as catch crops and green manure, the allelopathic properties of the cereals could now be much more extensively exploited, choosing varieties with optimal production of benzoxazinones and optimizing the time of sowing in relation to the formation of bioactive metabolites.     

Effect of exercise on activation of the p38 mitogen-activated protein kinase pathway, c-Jun NH2 terminal kinase, and heat shock protein 27 in equine skeletal muscle

OBJECTIVE: To investigate the effects of exercise on activation of mitogen-activated protein kinase (MAPK) signaling proteins in horses. ANIMALS: 6 young trained Standardbred geldings. PROCEDURE: Horses performed a 20-minute bout of exercise on a treadmill at 80% of maximal heart rate. Muscle biopsy specimens were obtained from the vastus lateralis and pectoralis descendens muscles before and after exercise. Amount of expression and intracellular location of phosphospecific MAPK pathway intermediates were determined by use of western blotting and immunofluorescence staining. RESULTS: Exercise resulted in a significant increase in phosphorylation of p38 pathway intermediates, c-Jun NH2 terminal kinase (JNK), and heat shock protein 27 (HSP27) in the vastus lateralis muscle, whereas no significant changes were found in phosphorylation of extracellular regulated kinase. In the pectoralis descendens muscle, phosphorylation of p38 and HSP27 was significantly increased after exercise. Immunohistochemical analysis revealed fiber-type- specific locations of phosphorylated JNK in type 2a/b intermediate and 2b fibers and phosphorylated p38 in type 1 fibers. Phosphorylated HSP27 was strongly increased after exercise in type 1 and 2a fibers. CONCLUSIONS AND CLINICAL RELEVANCE: The p38 pathway and JNK are activated in the vastus lateralis muscle after a single 20-minute bout of submaximal exercise in trained horses. Phosphorylation of HSP27 as detected in the study reported here is most likely induced through the p38 signaling pathway.     

Edge and area effects on avian assemblages and insectivory in fragmented native forests

Disentangling the confounded effects of edge and area in fragmented landscapes is a recurrent challenge for landscape ecologists, requiring the use of appropriate study designs. Here, we examined the effects of forest fragment area and plot location at forest edges versus interiors on native and exotic bird assemblages on Banks Peninsula (South Island, New Zealand). We also experimentally measured with plasticine models how forest fragment area and edge versus interior location influenced the intensity of avian insectivory. Bird assemblages were sampled by conducting 15?min point-counts at paired edge and interior plots in 13 forest fragments of increasing size (0.5?C141?ha). Avian insectivory was measured as the rate of insectivorous bird attacks on plasticine models mimicking larvae of a native polyphagous moth. We found significant effects of edge, but not of forest patch area, on species richness, abundance and composition of bird assemblages. Exotic birds were more abundant at forest edges, while neither edge nor area effects were noticeable for native bird richness and abundance. Model predation rates increased with forest fragmentation, both because of higher insectivory in smaller forest patches and at forest edges. Avian predation significantly increased with insectivorous bird richness and foraging bird abundance. We suggest that the coexistence of native and exotic birds in New Zealand mosaic landscapes enhances functional diversity and trait complementation within predatory bird assemblages. This coexistence results in increased avian insectivory in small forest fragments through additive edge and area effects.     

Copper-associated hepatic cirrhosis in a Friesian horse

A 6-year-old Friesian stallion was examined because of signs of exercise intolerance, stiff gait and symmetrical hind weakness, and increased serum liver enzymes. On presentation, the horse showed muscle atrophy of the hindquarters. Neurological investigation showed no abnormalities. Laboratory findings revealed a prolonged prothrombin time and increased levels of alkaline phosphatase (AF), aspartate aminotransferase (ASAT), gamma-glutamyl-transferase (GGT), lactate dehydrogenase (LDH), and bile acids. Histological evaluation of the liver revealed severe cirrhosis and intracytoplasmic greyish brown granules in almost all hepatocytes, sinusoidal Kuppfer cells, and macrophages. These granules stained strongly for copper. Treatment to slow hepatic fibrosis was advised and included oral prednisolone administration for at least 1 month. A diet to support liver function was formulated by a nutritional specialist, and vitamin E was advised as dietary supplement to support neuromuscular function. Soon after diagnosis, the animal showed signs of intravascular haemolysis, with the presence of Heinz bodies in peripheral blood smears, and haemoglobinuria. On the basis of this haemolytic crisis and the poor prognosis of the chronic hepatic disease, the horse was euthanized at the owners' request. Although we could not establish the cause of the hepatic copper accumulation, this case report highlights that excessive copper in the liver should be considered in the differential diagnosis of hepatic cirrhosis and Heinz body anaemia in the horse.     

An investigation on the presence of Chlamydiaceae in Swedish dogs

Background

Bacteria belonging to the family Chlamydiaceae cause a broad spectrum of diseases in a wide range of hosts, including man, other mammals, and birds. Upper respiratory and genital diseases are common clinical problems caused by Chlamydiaceae. Very little is known about chlamydial infections in dogs. Few clinical reports on natural disease in dogs describe mainly conjunctival and upper respiratory signs, and the role of Chlamydiaceae in genital disease is unclear. The present study aimed at studying the prevalence of Chlamydiaceae in healthy dogs and in dogs with genital or upper respiratory disease, including conjunctivitis.

Methods

A real-time polymerase chain reaction (PCR) for Chlamydiaceae was used to detect any chlamydial species within this family. Swab samples from the conjunctiva and the mucosal membranes of the oropharynx, rectum and genital tract were taken from 79 dogs: 27 clinically healthy dogs, 25 dogs with clinical signs from the genital tract and 28 dogs with conjunctivitis. There were 52 female and 27 male dogs. From 7 of the male dogs, additional semen samples were analysed.

Results

No Chlamydiaceae were detected from any dog.

Conclusions

Although the number of dogs that was included is limited, the results suggest that cases of Chlamydiaceae in dogs probably are related to infection from other species, and that dogs in general do not harbour Chlamydiaceae. Bacteria belonging to the family Chlamydiaceae do not seem to be of major importance for genital or ocular disease in Swedish dogs.     

Soil organic carbon–stock changes in Flemish grassland soils from 1990 to 2000

Total soil organic‐carbon (SOC) stocks for grassland soils in Flanders (N Belgium) were determined for the Kyoto Protocol reference year 1990 and 2000 in order to investigate whether these soils have been CO₂ sinks or sources during that period. The stocks were calculated by means of detailed SOC datasets, which were available at the community scale for the whole of Flanders. The total SOC stocks for Flemish grassland soils (1 m depth) were estimated at 38 Mt SOC in 1990 and 34 Mt SOC in 2000. The loss of SOC resulted from a decrease in the SOC content of grassland soils (71%) and could also partly (29%) be explained by a decline in grassland area. Significant decreases in %SOC for the 0–6 cm depth layer were found for the 1990s for the coarser‐textured soils with SOC losses ranging between –0.3% and –0.5% over the 10 y period. Specific management practices that disturb the SOC balance such as conversion to temporary grassland and a reduction of animal‐manure application are hypothesized to have contributed to the observed loss of SOC stocks. We furthermore conducted an analysis of uncertainty of the 1990 and 2000 grassland SOC–stocks calculation using Monte Carlo analysis. Probability‐distribution functions were determined for each of the inputs of the SOC‐stock calculation, enabling us to assess the uncertainty on the 1990 and 2000 SOC stocks. The frequency distributions of these simulated stocks both closely approached lognormal distributions, and their 95%‐confidence intervals ranged between 150% and 50% of the calculated mean SOC stock. The standard error on the measured decrease in SOC stocks in Flemish grassland soils during the 1990s was calculated to be 7–8 Tg SOC, which is equivalent to twice this decrease. This clearly shows that large‐scale changes in SOC stocks are uncertainty‐ridden, even when they are based on detailed datasets.     

Analysis of genetic diversity in a sweet potato (Ipomoea batatas L.) germplasm collection from Tanzania as revealed by AFLP

Sweet potato (Ipomoea batatas L.) is the fifth most important crop in the developing countries after rice, wheat, maize and cassava. The amplified fragment length polymorphism (AFLP) method was used to study the genetic diversity and relationships of sweet potato accessions in the germplasm collection of Sokoine University of Agriculture, Morogoro and Sugarcane Research Institute, Kibaha, Tanzania. AFLP analysis of 97 sweet potato accessions using ten primer combinations gave a total of 202 clear polymorphic bands. Each one of the 97 sweet potato accessions could be distinguished based on these primer combinations. Estimates of genetic similarities were obtained by the Dice coefficient, and a final dendrogram was constructed with the un-weight pair-group method using arithmetic average. AFLP-based genetic similarity varied from 0.388 to 0.941, with a mean of 0.709. Cluster analysis using genetic similarity divided the accessions into two main groups suggesting that there are genetic relationships among the accessions. Principal Coordinate analysis confirmed the pattern of the cluster analysis. Analysis of molecular variance revealed greater variation within regions (96.19%) than among regions (3.81%). The results from the AFLP analysis revealed a relatively low genetic diversity among the germplasm accessions and the genetic distances between regions were low. A maximally diverse subset of 13 accessions capturing 97% of the molecular markers diversity was identified. We were able to detect duplicates accessions in the germplasm collection using the highly polymorphic markers obtained by AFLP, which were found to be an efficient tool to characterize the genetic diversity and relationships of sweet potato accessions in the germplasm collection in Tanzania.