Gas chromatography for detection of citrus infestation by fruit fly larvae (Diptera: Tephritidae)

Tephritid fruit flies are serious economic pests worldwide. As larvae, they feed and develop within the pulp of host fruits, making infestation difficult to detect by visual inspection. At U.S. ports of entry, incoming produce shipments are checked for infestation by manually cutting open a small sample of fruit and searching for tephritid larvae. Consequently, there is a need for more sensitive, high-throughput screening methods. This study evaluated gas chromatography (GC) as a potential technology for improved detection of hidden infestation. Grapefruits (Citrus × paradisi Macfad.) infested with immature stages of the Caribbean fruit fly Anastrepha suspensa (Loew) (Diptera: Tephritidae) were examined to determine if infested fruit emitted a chemical profile distinct from that of non-infested fruit. Peaks identified by GC analysis were grouped into three classes. Chemicals detected in similar quantities in all samples, or slightly elevated in infested samples, were regarded as non-diagnostic background volatiles. Chemicals highly elevated after oviposition, during the last instar exit stage, and in experimentally-pierced fruit were interpreted to be indicators of citrus peel injury, and included d-limonene and β-ocimene. Chemicals elevated exclusively in the larval infestation stages were considered indicators of feeding damage and potentially diagnostic of infestation, and included hexyl butanoate and an unidentified compound. The peaks associated with injury and feeding were also detectable with a portable ultra-fast GC analyzer that required less than 80 s per sample. Further studies will investigate the potential application of these results for development of a rapid, non-destructive screening method for detection of tephritid infestation.     

Pathologic and immunohistochemical findings in an outbreak of systemic toxoplasmosis in a mob of red kangaroos

Toxoplasma gondii is a zoonotic protozoan pathogen that infects many endothermic vertebrates, including humans; the domestic cat and other felids serve as the definitive host. Macropodids are considered highly susceptible to toxoplasmosis. Here, we describe the clinical, pathologic, and immunohistochemical findings of an outbreak of systemic toxoplasmosis in a mob of 11 red kangaroos (Macropus rufus), with high morbidity (73%) and mortality (100%) rates. Affected animals had either severe and rapidly deteriorating clinical conditions or sudden death, which was correlated with widespread necrotizing lesions in multiple organs and intralesional T. gondii organisms identified via MIC3-specific immunohistochemistry and confirmed by REP529-specific rtPCR. Quantification of parasites demonstrated the highest parasite density in pulmonary parenchyma compared with other tissues. Our study highlights the continued importance of this severe condition in Australian marsupials.     

A faster inoculation assay for Armillaria using herbaceous plants

Armillaria (honey fungus) is a virulent necrotrophic pathogen that causes Armillaria root disease. Conventional Armillaria inoculation assays use young saplings as hosts and consequently are cumbersome, frequently conducted outdoors, and take many years from establishment to analysis of infection. We have developed and evaluated a faster inoculation assay for Armillaria that uses herbaceous plants as hosts, is carried out in controlled conditions, and reduces experimental durations to 3 months. Plant species of known susceptibility to Armillaria and comparisons between virulent A. mellea and opportunistic A. gallica were used to validate the assay. Mortality and diagnostic symptoms of Armillaria root disease such as epiphytic rhizomorphs and mycelial fans were used to assess levels of infection. We also attempted to reduce assay preparation time by substituting woody inocula with agar inocula, but typical symptoms of Armillaria root disease were only observed on plants infected with woody inocula. Through our assay, we identified five new potential herbaceous hosts of Armillaria: Kniphofia hirsuta, Hordeum vulgare, Lobelia cardinalis, Nicotiana tabacum and Helenium hoopesii – further expanding the extensive list of plants with susceptibility to Armillaria and suggesting infection of herbaceous species may be more widespread than currently acknowledged.     

Management strategies for forage rape (Brassica napus L. cv Goliath): Impact on dry-matter yield,plant reserves,morphology and nutritive value

Limited information is available on the grazing management principles of forage rape (Brassica napus L.), particularly in relation to grazing height and intensity and the impact of these on dry-matter (DM) yield and nutritive value. A glasshouse study was undertaken to investigate the effect of three defoliation heights (plant height at harvest; DH: 40, 70 and 90 cm; L, M and H DH respectively) and three defoliation intensities (height at which plants were cut; DI: 5, 20 and 35 cm of residual height; H, M and L DI respectively) on forage rape (cv Goliath) yield and nutritive value at two harvests (harvest 1, H1 and harvest 2, H2), and the impact of nitrogen (N) and water soluble carbohydrate (WSC) reserves on regrowth. Increasing DH from L to H increased estimated total DM yield (H1 plus H2) from 0.5 to 4.6 t DM/ha but DI did not affect yield. Dry-matter yield was optimized at 90 cm DH, but greater nutritive value was achieved by harvesting at lower levels of DH. Despite high in vitro DM digestibility (IVDMD; 852–889 g/kg), harvesting at 90 cm DH could not meet the protein requirement of lactating dairy cows and harvesting at lower levels risks nitrate poisoning. Our results indicate the optimum DH may be between 70 and 90 cm DH, and 20 and 35 cm DI, which requires further studies.     

Promotion of pistillate flowering inCucurbita by 2-chloroethylphosphonic acid

Summary All accessions tested of four cultivated species ofCucurbita responded to seedling treatment with 250 ppm 2-chloroethylphosphonic acid by developing only pistillate flowers for an extended period. This growth regulator is a promising means of facilitating hybrid seed production of squash. A similar treatment did not induce flowering of wild species ofCucurbita that were vegetative under the environmental conditions of the experiment.Approved by the Director of the New York State Agricultural Experiment Station for publication as Journal Paper No. 1737.     

Hypocalcemia in a critically ill patient

Objective: To present a case of clinical hypocalcemia in a critically ill septic dog. Case summary: A 12‐year old, female spayed English sheepdog presented in septic shock 5 days following hemilaminectomy surgery. Streptococcus canis was cultured from the incision site. Seven days after surgery, muscle tremors were noted and a subsequent low serum ionized calcium level was measured and treated. Intensive monitoring, fluid therapy, and antibiotic treatment were continued because of the sepsis and hypocalcemia, but the dog was euthanized 2 weeks after surgery. New or unique information provided: Low serum ionized calcium levels are a common finding in critically ill human patients, especially in cases of sepsis, pancreatitis, and rhabdomyolysis. In veterinary patients, sepsis or streptococcal infections are not commonly thought of as a contributing factor for hypocalcemia. Potential mechanisms of low serum ionized calcium levels in critically ill patients include intracellular accumulation of calcium ions, altered sensitivity and function of the parathyroid gland, alterations in Vitamin D levels or activity, renal loss of calcium, and severe hypomagnesemia. Pro‐inflammatory cytokines and calcitonin have also been proposed to contribute to low ionized calcium in the critically ill. Many veterinarians rely on total calcium levels instead of serum ionized calcium levels to assess critical patients and may be missing the development of hypocalcemia. Serum ionized calcium levels are recommended over total calcium levels to evaluate critically ill veterinary patients.