Recovery of brodifacoum in vomitus following induction of emesis in dogs that had ingested rodenticide bait

AIM: To assess the benefit of inducing emesis in dogs that have ingested rodenticide bait containing brodifacoum (BDF), by determining the amount of BDF in bait recovered from the vomitus relative to the estimated amount consumed.

METHODS: Between 2014 and 2015 samples of vomitus from seven dogs that ingested rodenticide baits containing BDF were submitted by veterinarians in New Zealand. All seven dogs had been given apomorphine by the veterinarian and vomited within 1 hour of ingesting the bait. Some or all of the bait particles were retrieved from each sample and were analysed for concentrations of BDF using HPLC. Based on estimations of the mass of bait consumed, the concentration of BDF stated on the product label, and the estimated mass of bait in the vomitus of each dog, the amount of BDF in the vomited bait was calculated as a percentage of the amount ingested.

RESULTS: For five dogs an estimation of the mass of bait ingested was provided by the submitting veterinarian. For these dogs the estimated percentage of BDF in the bait retrieved from the vomitus was between 10–77%. All dogs were well after discharge but only one dog returned for further testing. This dog had a normal prothrombin time 3 days after ingestion.

CONCLUSIONS AND CLINICAL RELEVANCE: The induction of emesis within 1 hour of ingestion can be a useful tool in reducing the exposure of dogs to a toxic dose of BDF. The BDF was not fully absorbed within 1 hour of ingestion suggesting that the early induction of emesis can remove bait containing BDF before it can be fully absorbed.     

Veränderungen der Vegetation durch experimentelle Erdgasbehandlung

Changes in vegetation caused by experimental leakage of natural gas The response of vegetation to natural gas leakage was studied in Central Bohemia, Czechoslovakia, in 1978–84. The gas used was of the following chemical composition: 93·8% methane, 3·6% ethane, 0·8% propane, 0·25% butane, 0·3% carbon dioxide. The test crops were planted in 60·5 m plots containing underground bores from which gas was released into the soil through distribution pipes. In addition, the response of some weed species was recorded. The important effects of contamination observed were restricted growth and decrease in the number of individuals present. A change in the green colour of leaves was recorded in Medicago sativa, Brassica oleracea var. acephala, Secale cereale, Lolium multiflorum. Beta vulgaris and Zea mays. An obvious shift of developmental stages was found in Solanum tuberosum. Helianthus annuus and Zea mays (delay of recruitment and blooming). Restricted or even failure of reproduction can be considered the typical effect of exposure to natural gas: this holds true especially for Allium cepa, Triticum aestivum, Hordeum distichon, Trifolium sp.div., Medicago sativa, and most of the weedy species. Various growth deformations, mostly found in the underground parts of the plants (tubers and roots in Solanum tuberosum, Beta vulgaris), can be ascribed to the unfavourable conditions of soil aeration. Plants of Chenopodium album and Medicago sativa growing in the vicinity of the gas release point had an increased number of stomata. There were changes in the course of the reflectance curves estimated in the population of Trifolium pratense subsp. sativum. There was a conspicuous decrease of the near infrared reflectance in the stand exposed to gas in comparison with the control. Decrease of coverage and density of the stands reduced species diversity and changes in the species' composition (reduction of sensitive species) are the general consequences of natural gas leakage. The first symptoms on the vegetation were observed after 15–30 days which corresponds to about 80–150 m³ of discharged gas.     

Nematicidal activity of essential oils and their components against the root-knot nematode

ABSTRACT Nematicidal activity of essential oils extracted from 27 spices and aromatic plants were evaluated in vitro and in pot experiments. Twelve of the twenty-seven essential oils immobilized more than 80% of juveniles of the root-knot nematode Meloidogyne javanica at a concentration of 1,000 mul/liter. At this concentration, most of these oils also inhibited nematode hatching. Essential oils of Carum carvi, Foeniculum vulgare, Mentha rotundifolia, and Mentha spicata showed the highest nematicidal activity among the in vitro tested oils. These oils and those from Origanum vulgare, O. syriacum, and Coridothymus capitatus mixed in sandy soil at concentrations of 100 and 200 mg/kg reduced the root galling of cucumber seedlings in pot experiments. The main components of these essential oils were tested for their nematicidal activity. Carvacrol, t-anethole, thymol, and (+)-carvone immobilized the juveniles and inhibited hatching at >125 mul/liter in vitro. Most of these components mixed in sandy soil at concentrations of 75 and 150 mg/kg reduced root galling of cucumber seedlings. In 3-liter pot experiments, nematicidal activity of the essential oils and their components was confirmed at 200 and 150 mg/kg, respectively. The results suggest that the essential oils and their main components may serve as nematicides.     

Parasitism of <Emphasis Type="Italic">Trichoderma</Emphasis> on <Emphasis Type="Italic">Meloidogyne javanica</Emphasis> and role of the gelatinous matrix

Trichoderma (T. asperellum-203, 44 and GH11; T. atroviride-IMI 206040 and T. harzianum-248) parasitism on Meloidogyne javanica life stages was examined in vitro. Conidium attachment and parasitism differed beween the fungi. Egg masses, their derived eggs and second-stage juveniles (J2) were parasitized by Trichoderma asperellum-203, 44, and T. atroviride following conidium attachment. Trichoderma asperellum-GH11 attached to the nematodes but exhibited reduced penetration, whereas growth of T. harzianum-248 attached to egg masses was inhibited. Only a few conidia of the different fungi were attached to eggs and J2s without gelatinous matrix; the eggs were penetrated and parasitized by few hyphae, while J2s were rarely parasitized by the fungi. The gelatinous matrix specifically induced J2 immobilization by T. asperellum-203, 44 and T. atroviride metabolites that immobilized the J2s. A constitutive-GFP-expressing T. asperellum-203 construct was used to visualize fungal penetration of the nematodes. Scanning electron microscopy revealed the formation of coiling and appressorium-like structures upon attachment and parasitism by T. asperellum-203 and T. atroviride. Gelatinous matrix agglutinated T. asperellum-203 and T. atroviride conidia, a process that was Ca²⁺-dependent. Conidium agglutination was inhibited by carbohydrates, including fucose, as was conidium attachment to the nematodes. All but T. harzianum could grow on the gelatinous matrix, which enhanced conidium germination. A biomimetic system based on gelatinous-matrix-coated nylon fibers demonstrated the role of the matrix in parasitism: T. asperellum-203 and T. atroviride conidia attached specifically to the gelatinous-matrix-coated fibers and parasitic growth patterns, such as coiling, branching and appressoria-like structures, were induced in both fungi, similarly to those observed during nematode parasitism. All Trichoderma isolates exhibited nematode biocontrol activity in pot experiments with tomato plants. Parasitic interactions were demonstrated in planta: females and egg masses dissected from tomato roots grown in T. asperellum-203-treated soil were examined and found to be parasitized by the fungus. This study demonstrates biocontrol activities of Trichoderma isolates and their parasitic capabilities on M. javanica, elucidating the importance of the gelatinous matrix in the fungal parasitism.     

Evaluation of Trichoderma spp. as a Biocontrol Agent Against Soilborne Fungi and Plant-parasitic Nematodes in Israel

The optimal conditions required to market Trichoderma as a biocontrol agent against soilborne fungi and nematodes are discussed. These include a proper formulation, an efficient delivery system, and alternative methods for Trichoderma's application.The implementation of Trichoderma in integrated pest management (IPM) can be achieved using a soil treatment which combines reduced amounts of biocides/fungicides and the Trichoderma preparation. Biocontrol activity can be increased by combining two (or more) types of biocontrol agents. Moreover, the construction of a genetically modified Trichoderma can lead to the improvement of certain traits which are absent or not highly expressed in the native microorganism isolated from its natural habitat.Different Trichoderma harzianum and T. lignorum isolates were tested for their nematicidal activity against the root-knot nematode Meloidogyne javanica. In short-term experiments, improved growth of nematode-infected plants and decreases in the root-galling index and the number of eggs per gram of root were achieved when nematode-infested soils were pre- exposed to the T. harzianum preparations. A long-term experiment resulted in improved growth and higher yield of nematode-infected plants, but no significant change in the galling index, either by pre-exposure of the fungus to the soil or by enrichment in the root-ball.As biocontrol is an integral part of the IPM philosophy, judicious use of Trichoderma against soilborne pathogens, when demonstrated to be consistently effective, practical and economic, can serve as a model for the introduction and implementation of other biocontrol means into IPM.     

Effects of Addition of Tissue‐Type Plasminogen Activator in In Vitro Fertilization Medium on Bovine Embryo Development and Quality

Plasminogen activators/Plasmin system plays pivotal role in regulating reproductive functions of mammals. Here, we examined the effects of modification of in vitro fertilization medium (IVF medium) with the addition of tissue‐type plasminogen activator (t‐PA), on bovine embryo development and quality, assessed by quantification of expression of various genes related to metabolism, oxidation, implantation and apoptosis. In addition, plasminogen activator activity (PAA) and plasminogen activator inhibition (PAI) were measured in the spent media. After conventional IVM, 2016 cumulus‐oocyte complexes (COCs) were divided into four groups with modified composition of the IVF medium containing t‐PA and/or its inhibitor epsilon‐aminocaproic acid (control, t‐PA, t‐PA+ε‐ACA, ε‐ACA). Presumptive zygotes were cultured for 8 days in synthetic oviductal fluid (SOF) medium; gene expression studies were carried out on morulae and blastocysts. t‐PA alone significantly suppressed cleavage and blastocyst formation rates, but this effect was neutralized by the addition of ε‐ACA. PAA in the treated group was significantly reduced by ε‐ACA, but without total elimination. Significant differences were detected in the expression of genes related to apoptosis and/or cell cycle arrest (BAX, BCL2L1, KAT2B) between embryos produced in t‐PA‐modified media and controls, giving an overall notion that the inferior developmental competence of treated embryos may be attributed to apoptotic phenomena induced by t‐PA. In conclusion, it appears that excessive t‐PA content in the IVF media, suppresses blastocyst formation rate, possibly due to induction of apoptotic phenomena.     

Enzyme-linked immunosorbent assay (ELISA) for the detection of spring viraemia of carp virus (SVCV) in tissue homogenates of the carp, Cyprinus carpio L.

Abstract. An enzyme-linked immunosorbent assay (ELISA) for the demonstration of the virus of spring viraemia of carp (SVCV) in liver, kidney and spleen homogenates, and in infected cell cultures is described. The sensitivity of the method is 10^2·8–10^3·5 TCID₅₀ 0·lml⁻¹ of the examined fluid. The specificity has been confirmed by the ELISA inhibition test and by results of virological examinations. Contamination with bacteria or fungi of samples taken from dead fish had no effect on the results of ELISA. Specific anti-SVCV sera were used successfully for the production of conjugates for the direct immunoperoxidase and immunofluorescence detection of SVCV in infected cell cultures.     

Abscisic acid, gibberellin and cell viability in cereal aleurone

The aleurone layer of cereals is a secretory tissue whose activity is regulated by abscisic acid (ABA) and gibberellins (GAs). Whereas GA triggers enzyme synthesis and secretion and initiates a program that culminates in cell death, ABA prevents enzyme production and cell death. Reactive oxygen species (ROS) are key players in regulating cell viability and GA sensitizes the aleurone cell to ROS. Sensitivity of GA-treated cells results in part from a reduction in steady-state amounts of mRNAs encoding enzymes that scavenge ROS. mRNAs encoding catalase, superoxide dismutase and ascorbate peroxidase are almost undetectable in aleurone layers 24 h after incubation in GA. For layers incubated in ABA, however, the amounts of these mRNAs increase. Western blotting and enzyme activity assays confirm that GA but not ABA reduced the amount and activity of ROS scavenging enzymes (Fath et al., 2001b). Substantial amounts of ROS are produced by enzymes engaged in lipid metabolism, and by the electron transport chain in the mitochondria. Aleurone layers contain abundant stores of triglycerides and ROS are produced as these lipids are rapidly converted to sugars. We hypothesize that the ROS produced in GA-treated aleurone cells bring about cell death by disrupting the plasma membrane. Aleurone cells incubated in ABA, on the other hand, are better able to maintain redox balance. ABA does not initiate rapid triglyceride metabolism, and the activities of ROS-scavenging enzymes remain high in ABA-treated cells. We conclude that GA initiates a metabolic cascade in aleurone cells that results in death from ROS. ABA maintains viability by keeping ROS under control. This revised version was published online in August 2006 with corrections to the Cover Date.