An easy and rapid method using microscopy to determine herbicide effects in Poaceae weed species

A new, easy, rapid and relatively inexpensive method using microscopy has been developed for the detection of herbicide effects in leaves of grass weed species displaying no visual signs of damage. The method has potential to be used as a tool to indicate future death of grass species due to herbicide exposure by observing phytochemical effects, i.e. early-warning effects. In the present study, Apera spica-venti (L.) Beauv., Bromus hordeaceus L., Alopecurus myosuroides Huds., Lolium perenne L. and Poa annua L. were exposed to lethal rates of four herbicides with different mode of action. The herbicides investigated were the thiocarbamate: prosulfocarb, the sulfonylurea: iodosulfuron, the aryloxyphenoxypropionate: fenoxaprop-P-ethyl and the organophosphate glyphosate. Autofluorescence of leaves was studied under a microscope using ultraviolet and blue light. The fluorescence of leaves treated to enhance flavonoids was also examined. To confirm the results, microspectrofluorometry was performed. Effects indicating future death of the grasses were observed in visually healthy leaves following treatment with prosulfocarb, glyphosate and iodosulfuron. No changes were detected following treatment with fenoxaprop-P-ethyl. After exposure to glyphosate or iodosulfuron, changes in the content of flavonoids and other compounds with a conjugation system and rigid structure and a decrease in the content of chlorophyll were detected in the leaves. Prosulfocarb treatment resulted in changes in the content of flavonoids and other compounds with a conjugation system and rigid structure and an increase in the content of chlorophyll in the leaves. The results obtained from microspectrofluorometry indicated that exposure to prosulfocarb caused a reduction in the flavonoids quercetin, naringenin and/or naringin.     

Upregulation of the PI3K/Akt Pathway in the Tumorigenesis of Canine Thyroid Carcinoma

Background

Information on the genetic events leading to thyroid cancer in dogs is lacking.

Hypothesis/Objectives

Upregulation of the PI3K/Akt pathway has an important role in the tumorigenesis of thyroid carcinoma in dogs.

Animals

Fifty‐nine dogs with thyroid carcinoma and 10 healthy controls.

Methods

Quantitative RT‐PCR was performed for VEGFR‐1, VEGFR‐2, EGFR, PIK3CA, PIK3CB, PDPK1, PTEN, AKT1, AKT2, COX‐2, and CALCA. Mutation analysis was performed for known hotspots of RAS (N, K, H), PIK3CA, BRAF, RET, and for the entire coding region of PTEN.

Results

Forty‐three dogs (73%) had follicular cell thyroid carcinoma (FTC) and 16 dogs (27%) had medullary thyroid carcinoma (MTC). The relative mRNA expressions of VEGFR‐1 (P < .001), VEGFR‐2 (P = .002), PDPK1 (P < .001), AKT1 (P = .009), and AKT2 (P < .001) were increased in FTC, and those of EGFR (P < .001), VEGFR‐1 (P = .036), and PIK3CA (P = .019) were increased in MTC when compared to normal thyroid glands. Mutation analysis of K‐RAS identified 2 activating missense mutations, which also have been described in thyroid cancer of humans. A G12R substitution was present in 1 FTC and an E63K substitution was present in 1 MTC. No functional mutations were found in the sequenced regions of H‐RAS, N‐RAS, PIK3CA, BRAF, RET, and PTEN.

Conclusions and Clinical Importance

The increased expression of several genes associated with PI3K/Akt signaling suggests the involvement of this pathway in the pathogenesis of thyroid carcinoma in dogs, warranting further research on pathway activation and gene amplification. The mutations most frequently associated with thyroid cancer in humans are rare in dogs.     

Clinical,Pathologic, and Immunohistochemical Prognostic Factors in Dogs with Thyroid Carcinoma

Background

Prognostic markers for dogs with thyroid tumors are limited.

Hypothesis/Objectives

To identify clinical, pathologic, and immunohistochemical prognostic factors for dogs with thyroid tumors.

Animals

Seventy dogs with thyroid neoplasia.

Methods

Retrospective study. Dogs with thyroid neoplasia were included when follow‐up information and formalin‐fixed paraffin‐embedded tumor samples were available. Immunohistochemistry (IHC) was performed for thyroglobulin, calcitonin, Ki‐67, and E‐cadherin. Correlation of tumor variables (diameter, volume, localization, scintigraphic uptake, thyroid function, IHC) with local invasiveness and metastatic disease was performed on all tumor samples. Forty‐four dogs treated by thyroidectomy were included in a survival analysis.

Results

Fifty dogs (71%) had differentiated follicular cell thyroid carcinoma (dFTC) and 20 (29%) had medullary thyroid carcinoma (MTC). At diagnosis, tumor diameter (P = .007; P = .038), tumor volume (P = .020), tumor fixation (P = .002), ectopic location (P = .002), follicular cell origin (P = .044), and Ki‐67 (P = .038) were positively associated with local invasiveness; tumor diameter (P = .002), tumor volume (P = .023), and bilateral location (P = .012) were positively associated with presence of distant metastases. Forty‐four dogs (28 dFTC, 16 MTC; stage I–III) underwent thyroidectomy. Outcome was comparable between dogs with dFTC and MTC. Macroscopic (P = .007) and histologic (P = .046) vascular invasion were independent negative predictors for disease‐free survival. Although time to presentation, histologic vascular invasion and Ki‐67 were negatively associated with time to metastases, and time to presentation was negatively associated with time to recurrence, no independent predictors were found. E‐cadherin expression was not associated with outcome.

Conclusions and Clinical Importance

Prognostic factors have been identified that provide relevant information for owners and clinicians.     

The protective immune response produced in dogs after primary vaccination with the LiESP/QA-21 vaccine (CaniLeish?) remains effective against an experimental challenge one year later

Control of canine leishmaniasis is an important objective for the benefit of dogs living in or visiting endemic areas and for public health because of the zoonotic nature of this disease. Resistance or susceptibility to developing canine leishmaniasis after exposure to Leishmania infantum is primarily determined by the ability of the immune system to develop an appropriate Th1-dominated specific response to the parasite. For this reason there is a need for effective canine vaccines that can decrease the number of dogs developing progressive infections. In this study, we followed the impact of the LiESP/QA-21 canine vaccine (composed of excreted-secreted proteins of L. infantum and the QA-21 saponin adjuvant), recently launched commercially in Europe, on selected humoral and cellular immune parameters following an infectious intravenous challenge with L. infantum promastigotes administered one year after the primary vaccine course. We also followed parasitological parameters to determine the parasitological status of the challenged dogs. In contrast to controls, vaccinated dogs retained significantly stronger cell-mediated immune responses against the parasite despite a virulent challenge and had significantly lower mean parasite burdens at the end of the study, associated with a lower probability of developing active infections. These results confirm that the immune responses generated by vaccination with LiESP/QA-21 are still effective against an intravenous challenge one year after the primary vaccine course.     

The SAHGA model to calculate the Spatial Ammoniacal Heterogeneity at the soil surface after fertiliser Granule Application

After dissolution of fertiliser granules, a high nitrogen concentration is recovered in the immediate vicinity of granules, which may enhance damaging processes like nitrite accumulation or ammonia volatilisation. Based on the diffusion equations of Cranck, the granule-soil microsite was modelled to obtain the actual fertilised surface plot and the effective rate of N application on this surface. Parameterisation of the diffusion coefficient of solutes consisted of a temperature and soil texture correction. The model was tested against an experimental data set obtained from soil incubations at two soil water contents (21.2% m³ m^–3 and 28.3% m³ m^–3) and two temperatures (4°C and 25°C) by comparing NH₄⁺ recovery at various distances from the granules. The simulated radius of the granule-soil microsite was more affected by the water content than by the temperature. The model is very accurate because 95–100% of total NH₄⁺ applied was recovered in the modelled surface depending on the experimental conditions (temperature and water content). The model was simple enough to be easily integrated into larger models dealing with surface-applied granule fertilisers.     

Evaluation of the in vivo anthelmintic properties of Mitragyna inermis (Willd.) as a livestock dewormer against parasitic hematophagous worm Haemonchus contortus infections in different breeds of lambs

Tropical Animal Health and Production - Gastrointestinal (GI) nematodes remain a major constraint on livestock production throughout the world. This study assessed the in vivo efficacy of the...     

Estimating illegal and unreported catches from marine ecosystems: a basis for change

To evaluate the impacts of fishing on marine ecosystems, the total extraction of fish must be known. Putting a figure on total extraction entails the difficult task of estimating, in addition to reported landings, discards, illegal and unmandated catches. Unreported catches cast various types of shadow, which may be tracked and estimated quantitatively. Some shadows of unreported catches are reviewed, for example, an innovative, well‐funded NGO publicizes illegal catch in the Southern Ocean. For various reasons, official figures often have the implicit but unacceptable assumption that such categories are null. We present an estimation procedure based on adjustment factors taken from observer reports, correspondents and published information that track changes in a regulatory regime, and hence reflect incentives and disincentives to misreport. Monte Carlo simulations address uncertainty using multiple sources of information to provide upper and lower estimates. Once in place, this method provides preliminary estimates that may be refined without disruption. The method is demonstrated for fisheries in Iceland and Morocco. We use a ‘by‐species’ approach for Icelandic cod and haddock, while the Moroccan catch is divided into demersal and pelagic categories. Results suggest that Icelandic cod catches may have been underestimated by between 1 and 14% at different times, and haddock by between 1 and 28%. Underestimation of Moroccan catches appears to have been as much as by 50%. These case studies show that it is possible to obtain estimates of misreporting, even when direct data are lacking. Our method encourages transparency because sources of information are presented so that uncertain values are easily identified, offering a basis for comment, collaboration and refinement in estimating illegal and unreported fishing.     

Characterization and quantification of proteins in lecithins

Several methods for extraction and quantification of proteins from lecithins were compared. Extraction with hexane-2-propanol-water followed by amino acid analysis is the most suitable method for isolation and quantification of proteins from lecithins. The detection limit of the method is 15 mg protein/kg lecithin, and the quantification limit is 50 mg protein/kg. The relative repeatability limits for samples containing 0-500 and 500-5000 mg protein/kg sample were 12.6 and 7.5%, respectively. The protein recovery ranged between 101 and 123%. The protein content has been determined in different kinds of lecithins. The results were as follows: standard soy lecithins (between 232 and 1338 mg/kg), deoiled soy lecithin (342 mg/kg), phosphatydylcholine-enriched soy lecithins (not detectable and 163 mg/kg), sunflower lecithins (892 and 414 mg/kg), and egg lecithin (50 mg/kg). The sodium dodecyl sulfate-polyacrylamide gel electrophoresis protein patterns of the standard soy and sunflower lecithins are very similar to those of soy flour. The protein profile of the egg lecithin shows several bands with a broad range of molecular masses. The molecular masses of the main proteins of soy lecithins and soy flour have been determined by matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) and ranged from 10.5 to 52.2 kDa. Most of the major proteins from soy and sunflower lecithins identified by MALDI-MS and electrospray tandem MS belong to the 11S globulin fraction, which is one of the main fractions of soy and sunflower seeds. In addition, the seed maturation protein P34 from the 7S globulin fraction of soy proteins has also been identified in soy lecithins. This protein has been reported as the most allergenic protein in soybean.