BackgroundDetermination of the interactions between hematophagous arthropods and their hosts is a necessary component to understanding the transmission dynamics of arthropod-vectored pathogens. Current molecular methods to identify hosts of blood-fed arthropods require the preservation of host DNA to serve as an amplification template. During transportation to the laboratory and storage prior to molecular analysis, genetic samples need to be protected from nucleases, and the degradation effects of hydrolysis, oxidation and radiation. Preservation of host DNA contained in field-collected blood-fed specimens has an additional caveat: suspension of the degradative effects of arthropod digestion on host DNA. Unless effective preservation methods are implemented promptly after blood-fed specimens are collected, host DNA will continue to degrade. Preservation methods vary in their efficacy, and need to be selected based on the logistical constraints of the research program.MethodsWe compared four preservation methods (cold storage at -20 °C, desiccation, ethanol storage of intact mosquito specimens and crushed specimens on filter paper) for field storage of host DNA from blood-fed mosquitoes across a range of storage and post-feeding time periods. The efficacy of these techniques in maintaining host DNA integrity was evaluated using a polymerase chain reaction (PCR) to detect the presence of a sufficient concentration of intact host DNA templates for blood meal analysis. We applied a logistic regression model to assess the effects of preservation method, storage time and post-feeding time on the binomial response variable, amplification success.ResultsPreservation method, storage time and post-feeding time all significantly impacted PCR amplification success. Filter papers and, to a lesser extent, 95 % ethanol, were the most effective methods for the maintenance of host DNA templates. Amplification success of host DNA preserved in cold storage at -20 °C and desiccation was poor.ConclusionsOur data suggest that, of the methods tested, host DNA template integrity was most stable when blood meals were preserved using filter papers. Filter paper preservation is effective over short- and long-term storage, while ethanol preservation is only suitable for short-term storage. Cold storage at -20 °C, and desiccation of blood meal specimens, even for short time periods, should be avoided.

Feeding upon vertebrate blood by mosquitoes permits transmission of diverse pathogens, including viruses, protozoa, and nematodes. Despite over a century of intensive study, no mosquito species is known to specialize on non-vertebrate hosts. Using molecular analyses and field observations, we provide the first evidence, to our knowledge, that a mosquito, Uranotaenia sapphirina, specializes on annelid hosts (earthworms and leeches) while its sympatric congener, Uranotaenia lowii, feeds only on anurans (frogs and toads). Our results demonstrate that Ur. sapphirina feeds on annelid hosts (100% of identified blood meals; n = 72; collected throughout Florida), findings that are supported by field observations of these mosquitoes feeding on Sparganophilus worms and freshwater leeches. These findings indicate that adult mosquitoes utilize a much broader range of host taxa than previously recognized, with implications for epidemiology and the evolution of host use patterns in mosquitoes.

Haemaphysalis longicornis Neumann (Asian longhorned tick) is an exotic and invasive tick species presenting a health and economic threat to the United States (U.S.) cattle industry due to its ability to transmit pathogens and infest hosts in large numbers. The objective of this study was to evaluate available products at causing H. longicornis mortality in a laboratory bioassay. The efficacy of products was evaluated at label rates using H. longicornis nymphs collected from a cattle farm in eastern Tennessee in two different bioassays (spray or dip) against untreated controls. After exposure, ticks were transferred to clean petri dishes and checked for mortality at 0, 1, 2, 3, 4, 21, 24, and 48 h post exposure. No mortality occurred in the untreated controls, whereas all treated ticks were dead within 24 h of exposure (P < 0.0001). These findings support the hypothesis that currently available spray and pour-on products are effective at causing H. longicornis mortality. We conclude that these acaricides can be used as a component to prevent H. longicornis dispersal and for control in the U.S.

The horn fly, Haematobia irritans irritans (L.), is one of the most important external parasites of cattle in North America and elsewhere. Horn fly adults have an intimate association with cattle, their primary host. With their often-high numbers and by feeding up to 38 times per day per fly, horn flies stress cattle. The resulting productivity loss is valued at more than 2.3 billion USD in the United States. Insecticides are commonly used to mitigate direct injury from feeding and indirect injury from disease transmission. This paper discusses horn fly biology, distribution, and management. Emphasis is on promising new approaches in novel insecticides, repellents, biological control, vaccines, animal genetics, and sterile insect technology that will lead to effective preventative tactics and the integration of smart technologies with horn fly management. We conclude with a discussion of research needs necessary to shift horn fly integrated pest management to an emphasis on preventative tactics and the precision use of reactive techniques.

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