Evaluation of microbial culture techniques for the isolation of Pythium insidiosum from equine tissues.

The purpose of this study was to evaluate the effects of sample handling, storage, and culture techniques on the isolation of Pythium insidiosum from infected equine tissues. Tissue and kunker samples obtained immediately posteuthanasia from a horse with subcutaneous pythiosis were used to assess the effects of sample type (kunkers vs. tissues), media type (selective vs. nonselective), storage technique, and storage time on P. insidiosum isolation rate. Overall, isolation rates were higher from fresh kunkers (94.6%) and stored kunkers (76.4%) than from fresh tissues (8.3%) or stored tissues (4.6%). Isolation of P. insidiosum also occurred more often on antibiotic-containing media than on nonselective media for both fresh and stored samples. For samples that were stored for 1-3 days prior to culture, P. insidiosum isolation rates were highest for the following techniques: kunkers stored at room temperature and plated on selective media (100%), kunkers stored at 4 C and then plated on either nonselective (91.7%) or selective (95.8%) media, kunkers stored on cold packs and then plated on either nonselective (93.8%) or selective (100%) media, kunkers stored in ampicillin solution and plated on selective media (100%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). For samples stored for 4-5 days, P. insidiosum isolation rates were highest for kunkers stored at 4 C and then plated on either nonselective (81.3%) or selective (87.5%) media, kunkers stored in ampicillin solution and then plated on selective media (87.5%), and kunkers stored in ampicillin/gentocin solution and plated on selective media (87.5%). Results of this study suggest that optimal isolation rates of P. insidiosum from infected equine tissues are achieved by culturing fresh kunkers on selective media. For samples that cannot be processed immediately, acceptable handling techniques include storage at room temperature for up to 3 days, refrigeration for up to 5 days, shipping on cold packs, and storage in antibiotic solution, each combined with subsequent inoculation on selective media.     

Potential uses for geographic information system-based planning and decision support technology in intensive food animal production

Although geographic information systems (GIS) have been used in many disciplines, the available technology in planning and decision support has only recently begun to be used in intensive animal production, in areas such as confined animal feeding operations. GIS-based planning and decision support systems have the potential to enhance many aspects of intensive animal production, such as disease monitoring and prevention, emergency management and nutrient waste disposal. Current uses of GIS in animal production are reviewed. Potential uses are illustrated using the example of the poultry industry on the Delmarva Peninsula, USA.     

Preliminary observations of genetic susceptibility of elk (Cervus elaphus nelsoni) to chronic wasting disease by experimental oral inoculation.

To compare the genetic susceptibility of elk (Cervus elaphus nelsoni) with various alleles of the PRNP gene, which encodes the normal cellular prion protein, to chronic wasting disease (CWD), eight 8-month-old elk calves of 3 genotypes (2 132MM, 2 132LM, and 4 132LL) were orally dosed with CWD-infected brain material from elk. During postinoculation (PI) month 23, both 132MM elk had lost appetite, developed clinical signs of weight loss and central nervous system (CNS) dysfunction, and were euthanized. Two other elk (both 132LM) developed similar clinical signs of disease and were euthanized during PI month 40. All 4 affected elk had microscopic lesions of spongiform encephalopathy (SE), and PrPres, the disease-associated form of the prion protein, was detected in their CNS and lymphoid tissues by use of immunohistochemical (IHC) and Western blot (WB) techniques. These findings indicate that elk with MM and LM at codon 132 are susceptible to orally inoculated CWD. All 4 LL elk are alive at PI year 4 and are clinically normal, which suggests that 132LL elk may have reduced susceptibility to oral infection with CWD-infected material or may have prolonged incubation time.     

Diagnosis and management of a retrobulbar abscess of periapical origin in a domestic rabbit.

Retrobulbar abscessation is the most common orbital disease of rabbits, and the lesions are notoriously difficult to treat successfully. This article describes the diagnosis, surgical treatment, and long-term management of an extensive abscess located within the right maxilla and retrobulbar space of a domestic rabbit. Preoperative assessment and long-term monitoring were achieved with a combination of radiography, ophthalmic ultrasonography, and CT. Extraoral rigid endoscopy via a fenestration in the maxilla was used to facilitate surgical debridement of the lesion, and extraction of multiple cheek teeth has been fundamental to achieving control. The long-term use of topical and systemic antibiotic preparations and nonsteroidal anti-inflammatory medication is discussed.     

Identification of Hammondia heydorni oocysts by a heminested-PCR (hnPCR-AP10) based on the H. heydorni RAPD fragment AP10

Toxoplasma gondii, Hammondia hammondi, Neospora caninum, Neospora hughesi and Hammondia heydorni are members of the Toxoplasmatinae sub-family. They are closely related coccidians with similarly sized oocysts. Molecular diagnostic techniques, especially those based on polymerase chain reaction (PCR), can be successfully applied for the differentiation of Hammondia-like oocysts. In this paper, we describe a rapid and simple method for the identification of H. heydorni oocysts among other members of the Toxoplasmatinae sub-family, using a heminested-PCR (hnPCR-AP10) based on a H. heydorni RAPD fragment available in molecular database. DNA of oocysts of H. heydorni yielded a specific fragment of 289-290 bp in the heminested-PCR assay. No product was yielded when the primers were used for the amplification of DNA extracted from T. gondii, N. caninum, N. hughesi and H. hammondi, thus allowing the differentiation of H. heydorni among other members of the Toxoplasmatinae sub-family. The hnPCR-AP10 was capable of detecting H. heydorni genetic sequences from suspensions with at least 10 oocysts. In conclusion, the hnPCR-AP10 proved to be a reliable method to be used in the identification of H. heydorni oocysts from feces of dogs.     

Contributions of marine infrastructures to marine planning and protected area networking

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Nutrient Contribution and Release Kinetics of Vermicompost Amended Pine Bark

The composting of organic waste materials using earthworms yields the value-added product vermicompost (VC). The use of VC as a growing substrate amendment for containerized horticultural crop production is a sustainable nutrient management approach which can benefit crop production by providing plant nutrients and improving crop growth. In order to provide nutrient management guidelines for users of VC amended container substrates, a study was conducted to quantify nutrients supplied to substrates solution and to develop nutrient release kinetic models to predict plant available nutrient based on total initial loading rates. Experimental treatments and results were expressed on a volume basis (4-L container) for easy interpretation and application by growers of crops in containers.     

Cherry picking by pseudomonads: After a century of research on canker,genomics provides insights into the evolution of pathogenicity towards stone fruits

Bacterial canker disease is a major limiting factor in the growing of cherry and other Prunus species worldwide. At least five distinct clades within the bacterial species complex Pseudomonas syringae are known to be causal agents of the disease. The different pathogens commonly coexist in the field. Reducing canker is a challenging prospect as the efficacy of chemical controls and host resistance may vary against each of the diverse clades involved. Genomic analysis has revealed that the pathogens use a variable repertoire of virulence factors to cause the disease. Significantly, strains of P. syringae pv. syringae possess more genes for toxin biosynthesis and fewer encoding type III effector proteins. There is also a shared pool of key effector genes present on mobile elements such as plasmids and prophages that may have roles in virulence. By contrast, there is evidence that absence or truncation of certain effector genes, such as hopAB, is characteristic of cherry pathogens. Here we highlight how recent research, underpinned by the earlier epidemiological studies, is allowing significant progress in our understanding of the canker pathogens. This fundamental knowledge, combined with emerging insights into host genetics, provides the groundwork for development of precise control measures and informed approaches to breed for disease resistance.     

Stress steroid levels and the short-term impact of routine dehorning in female southern white rhinoceroses (Ceratotherium simum simum)

Rhinoceros populations in Africa are under severe threat as a result of surging poaching rates and risk-mitigation strategies are continuously adapted in an attempt to ensure the survival of the species. This study compared faecal glucocorticoid metabolite (fGCM) levels of two age classes of limited free-ranging female white rhinos with fGCM levels of adult free-ranging female white rhinos. Subsequently, fGCM alterations in the limited free-ranging animals were monitored following routine dehorning as a measure of the animals’ short-term physiological stress response. Baseline fGCM levels differed significantly between tested groups, with both free-ranging and limited free-ranging adult animals showing significantly higher fGCM levels compared with limited free-ranging juvenile females. In contrast, baseline fGCM levels did not differ significantly between limited free-ranging and free-ranging adult individuals. Routine dehorning procedures resulted in a short-term stress response expressed by a significant increase in fGCM levels 48 h post-dehorning, with stress steroid levels returning to pre-dehorning concentrations 72 h after the procedure.