Prevalence of Anaplasma phagocytophilum in domestic felines in the United States

Anaplasma phagocytophilum is among the more common tick-borne disease agents in the United States. It is of veterinary and public health significance as dogs, cats, and human beings are known to be susceptible. A. phagocytophilum is transmitted trans-stadially by either nymphs or adults of either the black-legged tick (Ixodes scapularis) or the western black-legged tick (Ixodes pacificus). Little information is available regarding either the prevalence of this agent in cats or the dynamics of vector transmission. Four hundred and sixty feline blood samples from sites throughout the United States were assayed for antibodies to A. phagocytophilum using an indirect immunofluorescence assay (IFA). Results of the prevalence study showed that 20 samples (4.3%) were positive for A. phagocytophilum antibodies by IFA at a 1:50 dilution, however these results could not be confirmed by PCR analysis. PCR analysis for other cross-reacting Ehrlichia/Anaplasma spp. was also negative. These results demonstrate that natural infection of A. phagocytophilum in cats is uncommon.     

A role for thrombin receptor signaling in endothelial cells during embryonic development

The coagulation protease thrombin triggers fibrin formation, platelet activation, and other cellular responses at sites of tissue injury. We report a role for PAR1, a protease-activated G protein-coupled receptor for thrombin, in embryonic development. Approximately half of Par1-/- mouse embryos died at midgestation with bleeding from multiple sites. PAR1 is expressed in endothelial cells, and a PAR1 transgene driven by an endothelial-specific promoter prevented death of Par1-/- embryos. Our results suggest that the coagulation cascade and PAR1 modulate endothelial cell function in developing blood vessels and that thrombin's actions on endothelial cells-rather than on platelets, mesenchymal cells, or fibrinogen-contribute to vascular development and hemostasis in the mouse embryo.     

Mapping Infestations of Potato Cyst Nematodes and the Potential for Spatially Varying Application of Nematicides

The most important constraint to potato production in the UK is the damage caused by the potato cyst nematodes (PCN) Globodera pallida and Globodera rostochiensis. These are serious pests, capable of causing substantial yield loss. Modern management systems depend heavily on nematicides which, at c. £360 ha^–1 for granular and c. £550 ha^–1 for fumigant nematicides, are costly to use. Mapping field infestations of PCN gives growers the option of applying nematicide variably across their fields. We intensively sampled a field, infested with G. pallida, before and after potatoes were grown and used the results to consider decisions the grower might have taken and to examine the consequences of various actions. Sampling intensity is important in generating accurate maps. In our results, spatial independence in PCN counts occurred at about 60 m, although there was also evidence of spatial independence at a range of 10–20 m in intensively sampled areas of the field. A strategic requirement to keep PCN population densities small, rather than the more tactical objective of avoiding yield loss, would mean blanket treatment of infested fields with granular nematicide. Maps could then be used to target hot spots of PCN infestation for additional treatment with fumigant. This procedure would avoid blanket treatment with both types of nematicide, thereby diminishing the cost of chemicals applied and reducing possible environmental damage. However, the inverse relationship between pre-planting population density and multiplication rate of PCN makes it difficult to devise safe spatial application procedures, especially when the pre-planting population density is just less than the detection threshold.     

The effect of Johne's vaccination on tuberculin testing in farmed red deer (Cervus elaphus)

AIM: To assess the degree of interference with bovine tuberculin testing in farmed red deer that vaccination of young deer with an oil-adjuvanted vs aqueous formulation of live attenuated Mycobacterium paratuberculosis Strain 316F vaccines would be likely to cause, and to compare immunological responses between vaccine formulations.

METHODS: Five-month-old red deer (n=45) were randomly allocated to three treatment groups of 15 animals, which received either no vaccine, a single 2-ml dose of an oil-adjuvanted formulation or two 2-ml doses, 6 weeks apart, of an aqueous formulation of live attenuated M. paratuberculosis Strain 316F vaccine injected subcutaneously (S/C) in the neck (Control, Oil-adjuvant Ptb, and Aqueous Ptb groups, respectively). Injection-site reactions were described and measured on Weeks 3, 6 and 9. Animals were weighed and lymphocyte transformation tests (LTT) and antibody enzyme-linked immunosorbent assays (ELISA) using avian, bovine and Johnin tuberculin were conducted on blood samples collected at Weeks 0, 6, 12, 15, 24, 27, 36 and 39. A bovine mid-cervical skin test (MCT) was applied at Week 12, and comparative cervical skin tests (CCTs) at Weeks 24 and 36. At Week 42, the animals were slaughtered at a commercial deer slaughter premises and subjected to rigorous meat inspection.

RESULTS: Two animals were eliminated at the start of the trial due to a positive cross-reaction with bovine tuberculin in the initial LTT. Almost all animals reacted to the MCT at Week 12, with mean skin thicknesses of 3.9, 2.9 and 1.0 mm for the Oil- adjuvant Ptb, Aqueous Ptb and Control groups, respectively. When the CCT was conducted at Week 24, 2/15 Oil-adjuvant Ptb, 2/14 Aqueous Ptb and 1/14 Control animals were classified as CCT-positive to bovine tuberculin. By Week 36, all animals were CCT-negative. The Oil-adjuvant Ptb vaccination resulted in high persistent levels of antibody that reacted with bovine tuberculin, compared with negligible levels in the Aqueous Ptb group.

Overall, a single dose of the Oil-adjuvant Ptb vaccine in deer stimulated a vigorous, cross-reactive immune response, evidenced by high LTT, skin-test and antibody reactions to bovine tuberculin, with both cell-mediated and humoral characteristics. By comparison, two doses of the Aqueous Ptb vaccine produced less cross-reactivity and a bias towards a cell-mediated response. The Oil-adjuvant Ptb vaccine resulted in moderate injection- site lesions that were quite persistent, whereas the Aqueous Ptb vaccine resulted in smaller nodules that regressed more quickly.

CONCLUSIONS: Vaccination of farmed deer with an oil-adjuvanted Johne's vaccine has the potential to cause significant interference with routine tuberculin skin testing. The cross-reactivity should decline with time and the CCT should be able to clear MCT-positives, but there is a risk of false-positives to the blood test for tuberculosis (BTB), due to high persistent levels of antibody. The CCT could be used as a primary skin test in vaccinated deer on some farms.

The Aqueous Ptb caused fewer problems with skin testing and produced significantly less bovine antibody than the Oil-adjuvant Ptb, but stimulated persistent cell-mediated immune responses that may provide some protection against Johne's disease.     

Phenotypic and Genotypic Heterogeneity among Streptococcus iniae Isolates Recovered from Cultured and Wild Fish in North America,Central America and the Caribbean Islands

Abstract

Streptococcus iniae, the etiological agent of streptococcosis in fish, is an important pathogen of cultured and wild fish worldwide. During the last decade outbreaks of streptococcosis have occurred in a wide range of cultured and wild fish in the Americas and Caribbean islands. To gain a better understanding of the epizootiology of S. iniae in the western hemisphere, over 30 S. iniae isolates recovered from different fish species and geographic locations were characterized phenotypically and genetically. Species identities were determined biochemically and confirmed by amplification and sequencing of the 16S rRNA gene. Repetitive-element palindromic PCR fingerprinting as well as biochemical and antimicrobial susceptibility profiles suggest that a single strain of S. iniae was responsible for two different disease outbreaks among reef fishes in the Caribbean, one in 1999 and another in 2008. Interestingly, a majority of the isolates recovered from cultured fish in the Americas were genetically distinct from the Caribbean isolates and exhibited a trend toward higher minimal inhibitory concentration with respect to several antibiotics as well as greater genetic variability. The biological significance of this genetic variability is unclear, but it could have implications for future vaccine development and treatment.

Received April 20, 2014; accepted July 7, 2014     

Oviposition and Larval Habitat Preferences of the Saltwater Mosquito, Aedes vigilax, in a Subtropical Mangrove Forest in Queensland, Australia

Abstract Our aim was to investigate the oviposition and larval habitats of the saltwater mosquito Aedes vigilax (Skuse) (Diptera: Culicidae) in a mangrove forest system in subtropical Queensland, Australia. Eggshells (indicators of oviposition) and larvae were sampled in three habitat classes that were depicted in a schematic model. Two classes were in depressions or basins, either with hummocks or dense pneumatophore substrates, both of which retained water after tidal flooding. The third class was in freely flushed mangroves that corresponded with more frequent tidal connections than the depression classes. ANOVA and Tukey-Kramer tests were used to analyze the data. The null hypotheses were rejected: the hummock class was a significant habitat based on both eggshell and larval data. The conclusion was that mosquito production in the mangrove system was distributed unevenly between habitat classes, and that the hummock class had conditions suited to the requirements of the immature stages of Ae. vigilax. This research has the potential to inform mosquito management strategies by focusing treatment on the problem habitats and underpinning habitat modifications including reducing water retention in the basins.     

Commercial Evaluation of a Continuous Micronutrient Fortification Process for Nixtamal Tortillas

The corn tortilla plays an integral role in the Mexican diet and is an ideal vehicle for micronutrient fortification. Approximately 60% of corn tortillas in Mexico are produced from nixtamal, with the remainder prepared from masa flour. A process for continuous fortification of nixtamal tortillas was evaluated in two commercial mills in Mexico. A commercial powder dosifier was used to add micronutrient premix containing iron, zinc, folic acid, niacin, riboflavin, and thiamin to nixtamal (1 g/kg) as it was milled. After training and preliminary sampling, mills produced fortified tortillas unassisted for four weeks. Masa flow rates over a four‐day period were 10–12 kg/min in both plants. Premix flow from the dosifier showed good stability, with an average coefficient of variation of 1.6%. Initial results indicated consistency in the fortification process, with significantly increased variation during the four‐week production period. Fortified tortillas had significantly higher levels of all nutrients tested. Micronutrient losses were <11% in all cases except folic acid, which showed an 80% loss. Despite processing losses, fortification resulted in a nearly fivefold increase in folic acid compared with control tortillas. The new fortification process is technically viable and was well received by millers.