Evaluation of spectrophotometric and HPLC methods for shikimic acid determination in plants: models in glyphosate-resistant and -susceptible crops

Endogenous shikimic acid determinations are routinely used to assess the efficacy of glyphosate in plants. Numerous analytical methods exist in the public domain for the detection of shikimic acid, yet the most commonly cited comprise spectrophotometric and high-pressure liquid chromatography (HPLC) methods. This paper compares an HPLC and two spectrophotometric methods (Spec 1 and Spec 2) and assesses the effectiveness in the detection of shikimic acid in the tissues of glyphosate-treated plants. Furthermore, the study evaluates the versatility of two acid-based shikimic acid extraction methods and assesses the longevity of plant extract samples under different storage conditions. Finally, Spec 1 and Spec 2 are further characterized with respect to (1) the capacity to discern between shikimic acid and chemically related alicyclic hydroxy acids, (2) the stability of the chromophore (t1/2), (3) the detection limits, and (4) the cost and simplicity of undertaking the analytical procedure. Overall, spectrophotometric methods were more cost-effective and simpler to execute yet provided a narrower detection limit compared to HPLC. All three methods were specific to shikimic acid and detected the compound in the tissues of glyphosate-susceptible crops, increasing exponentially in concentration within 24 h of glyphosate application and plateauing at approximately 72 h. Spec 1 estimated more shikimic acid in identical plant extract samples compared to Spec 2 and, likewise, HPLC detection was more effective than spectrophotometric determinations. Given the unprecedented global adoption of glyphosate-resistant crops and concomitant use of glyphosate, an effective and accurate assessment of glyphosate efficacy is important. Endogenous shikimic acid determinations are instrumental in corroborating the efficacy of glyphosate and therefore have numerous applications in herbicide research and related areas of science as well as resolving many commercial issues as a consequence of glyphosate utilization.     

Prevalence of echocardiographic evidence of cardiac disease in apparently healthy cats with murmurs

The objective of this prospective study was to determine the prevalence of echocardiographic evidence of heart disease in apparently healthy cats with heart murmurs. Thirty-two privately owned domestic cats were examined. All cats were considered healthy on the basis of history and physical examination, except for the finding of a heart murmur on auscultation. Cats on any medications (besides regular flea, tick and heartworm preventative) or that were pregnant or lactating were excluded from this study. The prevalence of echocardiographic evidence of heart disease in this population of cats was 53%. Therefore, identification of a heart murmur on routine physical examination in apparently healthy cats warrants further investigation.     

Evaluation of Pet‐Related Management Factors and the Risk of Salmonella spp. Carriage in Pet Dogs from Volunteer Households in Ontario (2005–2006)

The purpose of this study was to determine pet‐related management factors that may be associated with the presence of Salmonella spp. in feces of pet dogs from volunteer households. From October 2005 until May 2006, 138 dogs from 84 households in Ontario were recruited to participate in a cross‐sectional study. Five consecutive daily fecal samples were collected from each dog and enrichment culture for Salmonella spp. was performed. A higher than expected number of the dogs (23.2%; 32/138) had at least one fecal sample positive for Salmonella, and 25% (21/84) of the households had at least one dog shedding Salmonella. Twelve serotypes of Salmonella enterica subsp. enterica were identified, with the predominant serotypes being Typhimurium (33.3%; 13/39), Kentucky (15.4%; 6/39), Brandenburg (15.4%; 6/39) and Heidelberg (12.8%; 5/39). Univariable logistic regression models were created with a random effect for household to account for clustering. Statistically significant risk factors for a dog testing positive included having contact with livestock, receiving a probiotic in the previous 30 days, feeding a commercial or homemade raw food diet, feeding raw meat and eggs, feeding a homemade cooked diet, and having more than one dog in the household. In two‐variable models that controlled for feeding raw food, the non‐dietary variables were no longer statistically significant. These results highlight the potential public health risk of including raw animal products in canine diets.     

Analysis of lidocaine and its major metabolite, monoethylglycinexylidide, in elk velvet antler by liquid chromatography with UV detection and confirmation by electrospray ionization tandem mass spectrometry

A sensitive liquid chromatographic (LC) method with UV detection was developed for the determination of residues of lidocaine (LID) and its major metabolite, monoethylglycinexylidide (MEGX), in elk velvet antler. The drugs were extracted from alkaline velvet antler homogenates, cleaned up on a C(18) solid-phase extraction cartridge, and separated on an Inertsil ODS-3 (3.0 x 250 mm, 5 microm) column using an isocratic mobile phase made up of 0.05 M phosphate buffer (pH 4.0)/acetonitrile (88:12, v/v) at a flow rate of 1.0 mL/min. The limits of quantification for LID and its major metabolite, MEGX, were 10 and 20 ng/g, respectively. The method was validated and used to measure the concentration of residues of LID and MEGX in elk velvet antlers harvested after either LID anesthesia or application of a drug-free control method (electro-anesthesia, EA). No LID or MEGX residues were detected in any of the antlers harvested after EA application. No MEGX residues were detected in any of the velvet antlers harvested after LID application, but residues of LID ranging in concentration from 68 to 4300 ng/g were detected in the three sections of the velvet antlers harvested after LID administration. LC-tandem mass spectrometry was used to confirm the presence of lidocaine detected in the velvet antlers.     

Management of 60 cases of diaphragmatic rupture

Sixty cases of diaphragmatic rupture in the dog and cat were reviewed. There was a 90 per cent success rate following surgical repair. There were six deaths; two of these resulted from pulmonary collapse and hydrothorax, two cats from pulmonary oedema, one cat with cardiac arrest and one due to hepatic necrosis. In none of the deaths was the presence or absence of chest drains implicated. Apart from cases of gastric dilation, all animals benefited from 12 to 24 hours cage rest and stabilisation before surgery. Simple anaesthetic and surgical guidelines for the management of diaphragmatic rupture are described.     

Effects of anabolic steroids on liver cell ultrastructure in sheep.

Castrated male sheep were sham inplanted or implanted with estradiol, trenbolone acetate or trenbolone acetate in combination with estradiol, and slaughtered after 56 days. Ultrastructural and morphometric examination of the livers of steroid-treated sheep showed an increase in the volume density of the cell occupied by rough endoplasmic reticulum and microbodies. In four of the eight sheep treated with trenbolone acetate, either alone or in combination with estradiol, enlarged mitochondria with crystalline inclusions were seen. The increase in hepatic rough endoplasmic reticulum may be of benefit to the sheep whereas the mitochondrial changes after anabolic steroid use indicate a cellular lesion of unknown significance for health.     

Virus infections in cattle and sheep in Scotland 1975-1978

Over the four-year period 1975-78 attempts were made to identify virus in samples from 1231 outbreaks of disease in cattle, and 682 outbreaks of disease in sheep. A virus identification was possible in 26 per cent of these disease outbreaks. The nature and significance of these virus infections is discussed.     

Identification of the ethylene precursor, 1-aminocyclopropane-1-carboxylic acid (ACC), in pollen

Within 15 min after application of pollen to the stigma of carnation flowers (Dianthus caryophyllus L. cultivar ‘White Sim’), ethylene (C₂H₄) production by the gynoecium had increased substantially. Pollen germination did not start until 1 h after pollination. Analysis of materials removed from pollen by a brief rinse with an aqueous solvent showed the presence of high concentrations of the C₂H₄ precursor, 1-aminocyclopropane-1-carboxylic acid (ACC). This compound, which was found to be present in pollen from a number of flower species and which increased in concentration as the anthers developed, may be an important mediator of the early response of flowers to pollination.