West Nile (WN) virus, a mosquito-transmitted virus native to Africa, Asia, and Europe, was isolated from two species of mosquitoes, Culex pipiens and Aedes vexans, and from brain tissues of 28 American crows, Corvus brachyrhynchos, and one Cooper's hawk, Accipiter cooperii, in Connecticut. A portion of the genome of virus isolates from four different hosts was sequenced and analyzed by comparative phylogenetic analysis. Our isolates from Connecticut were similar to one another and most closely related to two WN isolates from Romania (2.8 and 3.6 percent difference). If established in North America, WN virus will likely have severe effects on human health and on the health of populations of birds.

Using peptide nanoparticle technology, we have designed two novel vaccine constructs representing M2e in monomeric (Mono-M2e) and tetrameric (Tetra-M2e) forms. Groups of specific pathogen free (SPF) chickens were immunized intramuscularly with Mono-M2e or Tetra-M2e with and without an adjuvant. Two weeks after the second boost, chickens were challenged with 107.2 EID50 of H5N2 low pathogenicity avian influenza (LPAI) virus. M2e-specific antibody responses to each of the vaccine constructs were tested by ELISA. Vaccinated chickens exhibited increased M2e-specific IgG responses for each of the constructs as compared to a non-vaccinated group. However, the vaccine construct Tetra-M2e elicited a significantly higher antibody response when it was used with an adjuvant. On the other hand, virus neutralization assays indicated that immune protection is not by way of neutralizing antibodies. The level of protection was evaluated using quantitative real time PCR at 4, 6, and 8 days post-challenge with H5N2 LPAI by measuring virus shedding from trachea and cloaca. The Tetra-M2e with adjuvant offered statistically significant (P < 0.05) protection against subtype H5N2 LPAI by reduction of the AI virus shedding. The results suggest that the self-assembling polypeptide nanoparticle shows promise as a potential platform for a development of a vaccine against AI.

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West Nile fever outbreaks in the USA have caused over 700 human deaths, primarily due to neurological disease. The usual transmission route of West Nile virus (WNV) involves mosquito bites; however, alternative routes, including intrauterine infection, have also been reported. Here, the pathogenicity of WNV in mice during gestation has been investigated. An extremely high mortality rate was observed in pregnant mice (98 %, 60/61) compared with non-pregnant mice (52 %, 28/53; P<0.001), independent of the infecting dose or the week of pregnancy. Antibody titres were similar between pregnant and non-pregnant mice and between surviving and nonsurviving animals. WNV RNA titres in brains were also similar between pregnant and non-pregnant mice. WNV RNA could be detected in placentas and fetuses. These observations suggest strongly that, in the mouse model, pregnancy increases the risk of severe WNV infection and may help to understand the pathogenic mechanisms involved in WNV infection during pregnancy.