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.

ABSTRACTThe efficacies oftrans-cinnamaldehyde (TC) and eugenol (EG) for reducingSalmonella entericaserovar Enteritidis colonization in broiler chickens were investigated. In three experiments for each compound, 1-day-old chicks (n= 75/experiment) were randomly assigned to five treatment groups (n= 15/treatment group): negative control (-veS. Enteritidis, -ve TC, or EG), compound control (-veS. Enteritidis, +ve 0.75% [vol/wt] TC or 1% [vol/wt] EG), positive control (+veS. Enteritidis, -ve TC, or EG), low-dose treatment (+veS. Enteritidis, +ve 0.5% TC, or 0.75% EG), and high-dose treatment (+veS. Enteritidis, +ve 0.75% TC, or 1% EG). On day 0, birds were tested for the presence of any inherentSalmonella(n= 5/experiment). On day 8, birds were inoculated with ∼8.0 log10CFUS. Enteritidis, and cecal colonization byS. Enteritidis was ascertained (n= 10 chicks/experiment) after 24 h (day 9). Six birds from each treatment group were euthanized on days 7 and 10 after inoculation, and cecalS. Enteritidis numbers were determined. TC at 0.5 or 0.75% and EG at 0.75 or 1% consistently reduced (P< 0.05)S. Enteritidis in the cecum (≥3 log10CFU/g) after 10 days of infection in all experiments. Feed intake and body weight were not different for TC treatments (P> 0.05); however, EG supplementation led to significantly lower (P< 0.05) body weights. Follow-upin vitroexperiments revealed that the subinhibitory concentrations (SICs, the concentrations that did not inhibitSalmonellagrowth) of TC and EG reduced the motility and invasive abilities ofS. Enteritidis and downregulated expression of the motility genesflhCandmotAand invasion geneshilA,hilD, andinvF. The results suggest that supplementation with TC and EG through feed can reduceS. Enteritidis colonization in chickens.

Tuberculosis is one of the important public health problems in Egypt. However, limited information on the Mycobacterium tuberculosis genotypes circulating in Egypt is available. A total of 151 M. tuberculosis strains were characterized by spoligotyping. The results revealed that 74.8% of M. tuberculosis isolates grouped into 13 different clusters, while 25.2% had unique spoligotype patterns. Comparison with an international spoligotyping database (the SITVIT2 database) showed that types SIT53 (T1 variant) and SIT54 (Manu2 variant) were the most common types between cluster groups. In addition, new shared types SIT2977, SIT2978, and SIT2979 were observed. The results identified for the first time an unusually high proportion of ancestral Manu strains of M. tuberculosis from patients in Egypt. The percentage of the Manu clade in this study (27.15%) was significantly higher than its overall representation of 0.4% in the SITVIT2 database. We show that in Egypt tuberculosis is caused by a predominant M. tuberculosis genotype belonging to the ancestral Manu lineage which could be a missing link in the split between ancestral and modern tubercle bacilli during the evolution of M. tuberculosis.Despite the availability of antituberculosis (anti-TB) drugs, the disease burden of human TB remains a very serious and widespread public health problem. Many of the 22 countries most affected by human TB identified by the World Health Organization (WHO) are developing countries (11). Although Egypt is not on the WHO list of 22 high-TB-burden countries, it is considered one of the high-burden countries in WHO's Eastern Mediterranean region, where TB constitutes the second most important public health problem after schistosomiasis (21). Every year, the National Tuberculosis Control Program of the Ministry of Health and Population (MOHP) registers over 12,000 new TB patients; more than 50% of the cases are sputum smear-positive pulmonary TB. On the basis of an annual risk of infection of 0.32%, it is estimated that about 8,000 people receive a diagnosis of TB at facilities other than those of MOHP (25).In the context described above, it is important to have a precise picture of the predominant and emerging Mycobacterium tuberculosis clones in Egypt, in order to have a first snapshot of the epidemiology of TB. The advent of molecular typing methods has given a new boost to epidemiological studies of TB in recent years (3); the most commonly used methods are IS6110-based restriction fragment length polymorphism (RFLP) (IS6110-RFLP) and spoligotyping (12,22). As opposed to IS6110-RFLP, which was considered the "gold standard" for the genotyping of M. tuberculosis in the early 1990s (22) and which is time-consuming and labor-intensive and requires large amounts of highly purified DNA, PCR-based spoligotyping avoids the problems associated with the slow growth of these bacteria, thereby offering the possibility of the rapid recognition of outbreaks (2,4,12). The clinical usefulness of spoligotyping is determined by its rapidity both in d...

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