United States Department of Agriculture-Agricultural Research Service research on pre-harvest prevention of mycotoxins and mycotoxigenic fungi in US crops

Mycotoxins (ie toxins produced by molds) are fungal metabolites that can contaminate foods and feeds and cause toxic effects in higher organisms that consume the contaminated commodities. Therefore, mycotoxin contamination of foods and feeds results is a serious food safety issue and affects the competitiveness of US agriculture in both domestic and export markets. This article highlights research accomplished by Agricultural Research Service (ARS) laboratories on control of pre-harvest toxin contamination by using biocontrol, host-plant resistance enhancement and integrated management systems. Emphasis is placed on the most economically relevant mycotoxins, namely aflatoxins produced by Aspergillus flavus, Link, trichothecenes produced by various Fusarium spp and fumonisins produced by F verticillioides. Significant inroads have been made in establishing various control strategies such as development of atoxigenic biocontrol fungi that can outcompete their closely related, toxigenic cousins in field environments, thus reducing levels of mycotoxins in the crops. Potential biochemical and genetic resistance markers have been identified in crops, particularly in corn, which are being utilized as selectable markers in breeding for resistance to aflatoxin contamination. Prototypes of genetically engineered crops have been developed which: (1) contain genes for resistance to the phytotoxic effects of certain trichothecenes, thereby helping reduce fungal virulence, or (2) contain genes encoding fungal growth inhibitors for reducing fungal infection. Gene clusters housing the genes governing formation of trichothecenes, fumonisins and aflatoxins have been elucidated and are being targeted in strategies to interrupt the biosynthesis of these mycotoxins. Ultimately, a combination of strategies using biocompetitive fungi and enhancement of host-plant resistance may be needed to adequately prevent mycotoxin contamination in the field. To achieve this, plants may be developed that resist fungal infection and/or reduce the toxic effects of the mycotoxins themselves, or interrupt mycotoxin biosynthesis. This research effort could potentially save affected agricultural industries hundreds of millions of dollars during years of serious mycotoxin outbreaks.     

BSE and scrapie diagnosis in Germany

The detection of pathological prion protein is considered as pathognomonic for the diagnosis of transmissible spongiform encephalopathies. According to the EU regulations cattle older than 30 months of age (Germany 24 months) and slaughtered for human consumption must be tested by using BSE rapid tests. Likewise must be fallen stock and clinically affected animals. This article gives an overview over the diagnostic hierarchy and organization of the diagnostic system for BSE and scrapie in Germany. All suspect cases found by rapid testing are reinvestigated and clarified by the National reference laboratory for these diseases which is part of the recently founded Institute of Novel and Emerging Infectious Diseases at the Federal Research Centre for Virus Diseases of Animals located on the isle of Riems. Until the end of 2001 130 BSE cases were confirmed out of 230 submissions.     

A comparison of 2 different suture patterns for skin closure of canine ovariohysterectomy

The purpose of this study was to compare postoperative wound healing in canine ovariohysterectomy following the use of an absorbable monofilament poliglecaprone 25 suture in 2 different skin closure techniques, the buried continuous subcuticular (BCS) suture pattern and the simple interrupted (SI) suture pattern. These 2 skin closure techniques were evaluated against a nonabsorbable polypropylene monofilament suture in an SI pattern. Wounds were assessed by using a semiquantitative scoring system at 18 to 24 hours and 10 to 14 days, postoperatively. Results indicated that the BCS closure using poliglecaprone 25 demonstrated a higher rate of tissue reactivity initially (18-24 hours postoperatively), as compared with the SI closure using either suture material. By 10 to 14 days postoperatively, poliglecaprone 25 used in a BCS closure was associated with significantly lower wound scores than was the same material used in an SI closure. It was concluded that the BCS closure may effect a better cosmetic appearance to the skin closure in a canine ovariohysterectomy at the time of the recheck appointment. Furthermore, by obviating the need for suture removal, use of the BCS pattern may eliminate the requirement for this return appointment.     

Closure of median sternotomy in dogs: suture versus wire

Twenty normal, large-breed dogs underwent median sternotomy. Median sternotomies were closed with 20-gauge orthopedic wire in 10 dogs and no. 2 polybutester in 10 dogs. Closure with suture was faster than with wire (6.7 +/- 1.8 minutes versus 9.1 +/- 1.9 minutes, respectively). Significant differences were not observed in degree of postoperative pain or wound complication rates. Sternotomies closed with wire showed a trend to be more stable and had significantly less displacement on radiographic evaluation at 28 days. All sterna closed with wire examined histopathologically showed evidence of chondral or osteochondral bridging, while sterna closed with suture only showed fibrous union.     

A double-blind,randomized, placebo-controlled study of pimobendan in dogs with dilated cardiomyopathy

A double-blind, randomized, placebo-controlled study was conducted to examine the effect on heart failure class and survival of pimobendan, an oral calcium-sensitizing inodilator, in dogs with dilated cardiomyopathy (DCM). Pimobendan (0.3-0.6 mg/kg body weight/d) or placebo was administered to English Cocker Spaniels (CSs; n = 10) and Doberman Pinschers (DPs: n = 10) that had DCM in addition to background therapy of furosemide, enalapril, and digoxin. Addition of pimobendan to standard triple therapy was associated with a significant improvement in heart failure class, regardless of breed (P < .02, Mann-Whitney rank sum test). Overall, 8 of 10 animals in the pimobendan-treated group, and 1 of 10 animals in the placebo group improved their heart failure status by at least I modified New York Heart Association functional class after initial stabilization (P = .005, Fisher's exact test). Pimobendan had no significant effect on survival in the CSs (P = 0.77, log-rank test), but DPs treated with pimobendan had significantly longer survival times compared with placebo (P < .02, log-rank test), with a median survival time of 329 days in the pimobendan group compared with 50 days in the placebo group, and a hazard ratio of 3.4 (95% confidence interval 1.4-39.8). Pimobendan resulted in significant improvement in heart failure class when added to standard therapy in this group of dogs with DCM, and may have contributed to improved survival in DPs.     

Use of the Cell-Dyn 3500 to predict leukemic cell lineage in peripheral blood of dogs and cats

Background— Morphology and cytochemistry are the foundation for classification of leukemias in dogs and cats. Advances in automated hematology instrumentation, immunophenotyping, cytogenetics, and molecular biology are significantly improving our ability to recognize and classify spontaneous myeloproliferative and lymphoproliferative disorders. Objective— The purpose of this study was to assess the utility of flow cytometry‐based light scatter patterns provided by the Cell‐Dyn 3500 (CD3500) automated hematology analyzer to predict the lineage of leukemic cells in peripheral blood of dogs and cats. Methods— Leukemic cells from 15 dogs and 6 cats were provisionally classified using an algorithm based on the CD3500 CBC output data and were subsequently phenotyped by enzyme cytochemistry, immunocytochemistry, indirect flow cytometry, and analysis of antigen receptor gene rearrangement. Results— The algorithm led to correct predictions regarding the ontogeny of the leukemic cells (erythroid/megakaryocytic potential, myeloid leukemia, monocytic leukemia, chronic granulocytic leukemia, lymphoid leukemia) in 19/21 animals. Mismatches in the WBC impedance count and the WBC optical count in conjunction with microscopic assessment of blasts in the blood were useful for predicting myeloproliferative disorders with erythroid or megakaryocytic potential. The leukocyte light scatter patterns enabled distinction among myeloid leukemias (represented by acute myelomonocytic leukemia, acute monocytic leukemia, chronic granulocytic leukemia) and lymphocytic leukemias (including acute and chronic lymphocytic leukemias). One case of acute lymphocytic leukemia was misidentified as chronic lymphocytic leukemia. Conclusions— Algorithmic analyses can be applied to data generated by the CD3500 to predict the ontogeny of leukemic cells in the peripheral blood of dogs and cats. This rapid and quantitative technique may be used to improve diagnostic decisions, expand therapeutic choices, and increase prognostic accuracy.     

Molecular Variability Within Diaporthe/Phomopsis helianthi from France

ABSTRACT Diaporthe/Phomopsis helianthi causes brown stem canker of sunflower (Helianthus annuus) and is responsible for considerable yield loss. This species shows considerable variation for morphological characters, growth, and pathogenicity. Molecular variability of two sample groups was assessed with amplified fragment length polymorphism (AFLP) markers. Isolates of the first sample were collected from infected sunflower tissues from the main regions in France where the crop is grown, whereas isolates from the second sample came from stems within a single field of sunflower. A soybean strain was taken as an outgroup for AFLP analyses. Within sample one, the greatest genetic distance among isolates was 0.97, whereas it was 0.44 within sample two isolates. For the whole of France, the average genetic distance was 0.68, whereas in the one field it was 0.12. Nei's genetic diversity indices were 0.20 and 0.06 for France and for one field, respectively. The greatest genetic distance was found between isolates from the most northern crops. The greatest genetic distance between D. helianthi isolates and the strain isolated from soybean was similar to that observed for D. helianthi isolates from different geographical areas. The problems in defining the genus Phomopsis are discussed. It is shown that internal transcribed spacer sequencing could be a useful criteria for Diaporthe/Phomopsis species determination. The considerable genetic variability of the pathogen could lead to the occurrence of new strains that could be more aggressive or more resistant to chemical control.