Mohammad Q. Al-Natour

Replication and transmission of H9N2 influenza viruses in ferrets: evaluation of pandemic potential.

H9N2 avian influenza A viruses are endemic in poultry of many Eurasian countries and have caused repeated human infections in Asia since 1998. To evaluate the potential threat of H9N2 viruses to humans, we investigated the replication and transmission efficiency of H9N2 viruses in the ferret model. Five wild-type (WT) H9N2 viruses, isolated from different avian species from 1988 through 2003, were tested in vivo and found to replicate in ferrets. However these viruses achieved mild peak viral titers in nasal washes when compared to those observed with a human H3N2 virus. Two of these H9N2 viruses transmitted to direct contact ferrets, however no aerosol transmission was detected in the virus displaying the most efficient direct contact transmission. A leucine (Leu) residue at amino acid position 226 in the hemagglutinin (HA) receptor-binding site (RBS), responsible for human virus-like receptor specificity, was found to be important for the transmission of the H9N2 viruses in ferrets. In addition, an H9N2 avian-human reassortant virus, which contains the surface glycoprotein genes from an H9N2 virus and the six internal genes of a human H3N2 virus, showed enhanced replication and efficient transmission to direct contacts. Although no aerosol transmission was observed, the virus replicated in multiple respiratory tissues and induced clinical signs similar to those observed with the parental human H3N2 virus. Our results suggest that the establishment and prevalence of H9N2 viruses in poultry pose a significant threat for humans.

Effect of Different Levels of Maternally Derived Antibodies on Protection Against Infectious Bursal Disease Virus

Abstract Fertile eggs were obtained from three different broiler breeder flocks with different levels of virus neutralizing antibodies to infectious bursal disease virus. Egg yolk from these flocks was tested for antibody titers by the virus neutralization test. Flock I eggs had no antibodies, flock II had medium level antibodies (1:200–1600; geometric mean = 1:975), and flock III had a high level of antibodies (1:1600–6400; geometric mean = 1:3365). Chicks from the above flocks were challenged each with 102 50% embryo infective dose of the IN serotype 1 variant virus at 1, 2, and 4 wk of age and examined at 5 and 11 days postchallenge. The average organ/body weight ratios were calculated and statistically analyzed. Chicks with no maternal antibodies were not protected at any age. Chicks with medium levels of maternal antibodies were protected when challenged at 1 and 2 wk of age. Chicks with high levels of maternally derived antibodies were protected when challenged at all the ages tested. The above results were statistically significant (P < 0.05).

Interspecies and intraspecies transmission of triple reassortant H3N2 influenza A viruses

1. AbstractThe triple reassortant H3N2 viruses were isolated for the first time from pigs in 1998 and are known to be endemic in swine and turkey populations in the United States. In 2004, we isolated two H3N2 triple reassortant viruses from two turkey breeder flocks in Ohio and Illinois. Infected hens showed no clinical signs, but experienced a complete cessation of egg production. In this study, we evaluated three triple reassortant H3N2 isolates of turkey origin and one isolate of swine origin for their transmission between swine and turkeys. Although all 4 viruses tested share high genetic similarity in all 8 genes, only the Ohio strain (A/turkey/Ohio/313053/04) was shown to transmit efficiently both ways between swine and turkeys. One isolate, A/turkey/North Carolina/03, was able to transmit from pigs to turkeys but not vice versa. Neither of the other two viruses transmitted either way. Sequence analysis of the HA1 gene of the Ohio strain showed one amino acid change (D to A) at residue 190 of the receptor binding domain upon transmission from turkeys to pigs. The Ohio virus was then tested for intraspecies transmission in three different avian species. The virus was shown to replicate and transmit among turkeys, replicate but does not transmit among chickens, and did not replicate in ducks. Identifying viruses with varying inter- and intra-species transmission potential should be useful for further studies on the molecular basis of interspecies transmission.

Flock-level prevalence of Eimeria species among broiler chicks in northern Jordan

Six chicks (3-6 weeks of age) were taken randomly from each of 200 broiler farms in northern Jordan, these chicks were submitted for post-mortem and parasitological examinations. Seven Eimeria spp. were identified: E. acervulina, E. brunetti, E. maxima, E. necatrix, E. mivati, E. mitis, and E. tenella. Half (50%) of the farms surveyed had all six chicks infected, 23% of the farms were free of the infection. E. tenella was the most prevalent species (39%) followed by E. necatrix (12%), E. brunitti (12%), and E. maxima (10%). Prevalences did not vary by flock size. Also, neither the use of coccidiostat nor previous coccidiosis clinical outbreaks was associated with the prevalence of coccidiosis.

PATHOGENICITY, ATTENUATION, AND IMMUNOGENICITY OF INFECTIOUS BURSAL DISEASE VIRUS

An investigation was conducted in specific-pathogen-free chickens on the pathogenicity of bursa-derived and tissue culture-attenuated classic (STC) and variant (IN) serotype 1 strains of infectious bursal disease virus. The IN bursa-derived virus caused bursal inflammation, necrosis, and atrophy earlier than the bursa-derived STC virus. Both viruses lost their pathogenicity after four passages in BGM-70 cells. A statistically significant level (P < 0.05) of protection was observed in SPF chickens vaccinated with the attenuated IN virus used as a live or inactivated vaccine followed by homologous (IN) and heterologous (STC) challenge with bursa-derived viruses.

Genetic Comparison of H9N2 AI Viruses Isolated in Jordan in 2003

Abstract During the 1990s, several outbreaks of avian influenza (AI), caused by viruses of the H9N2 subtype, were reported in poultry in various parts of the world. Currently, this infection seems to be endemic in poultry in the Middle and Far East, and the extensive circulation of H9N2 in poultry represents a risk factor for infection of humans, because viruses of this subtype have been sporadically introduced into the human population. Little is known about the gene constellation of the H9N2 viruses that are currently circulating in the Middle East; thus, gene sequences of eight IA viruses of the H9N2 subtype isolated in Jordan in 2003 from poultry were analyzed. The results of this investigation show that all eight Jordanian isolates are closely related to each other and to other H9N2 isolates from the Middle East. Seven of eight genes of the Jordanian strains show a percentage of homology not higher than 95% with the genes of two H9N2 strains, A/HK/1073/99 and A/HK/1074/99, isolated from humans in Hong Kong. The M gene is more closely related to the corresponding gene of the two H9N2 human isolates from Hong Kong (97.7–98.2% homology). This homology suggests that the M gene of the Jordanian and human Hong Kong strains could derive from a common ancestor.

Interference Between Mild and Pathogenic Strains of Infectious Bursal Disease Virus in Chickens

Abstract Infectious bursal disease virus is a contagious, immunosuppressive disease of young chickens that is controlled by vaccination. Cross-protection occurs between different strains of the virus as a result of shared neutralizing epitopes. However, interactions between two antigenically similar strains (a mild and a pathogenic) coinfecting the same host have not been investigated. Groups of specific-pathogen-free chickens were inoculated with a mild strain followed by a pathogenic strain at 0, 16, 24, or 48 hr postinoculation (PI) with a mild strain. Virus persistence and the predominant strain of the virus were determined by reverse transcriptase–polymerase chain reaction and restriction fragment length polymorphism analysis, respectively, in bursas at 2, 4, 8, 14, and 21 days PI with the pathogenic strain. Severity of infection was assessed by the bursa/body weight ratios and histopathologic lesion scores. The mild virus interfered with replication of the pathogenic virus. The greatest interference was observed when the pathogenic strain was inoculated 24 hr PI with the mild strain. The interference phenomenon observed might be due to competition for host receptor sites or production of cytokine(s). This interference phenomenon could have practical implications for vaccine usage and protection.

Weighted dual functions for Bernstein basis satisfying boundary constraints

In this paper, we consider the issue of dual functions for the Bernstein basis which satisfy boundary conditions. The Jacobi weight function with the usual inner product in the Hilbert space are used. Some examples of the transformation matrices are given. Some figures for the weighted dual functions of the Bernstein basis with respect to the Jacobi weight function satisfying boundary conditions are plotted. We discuss special cases of the Jacobi weight function as the Legendre weight function and the Chebyshev weight functions of the first, second, and third kinds.

Molecular characterization of a recent Newcastle disease virus outbreak in Jordan.

Newcastle disease (ND) is a highly contagious viral disease and is a continuous threat to the poultry industry worldwide. In the early months of 2011, several devastating ND outbreaks occurred in Jordan affecting broilers, layers and breeders. The fusion gene of the isolated Newcastle disease virus (NDV) was partially amplified by RT-PCR, then directly sequenced. The NDV isolates were found to have the motif112RRQKRF117. This motif and a mean death time (MDT) of 46 h are indicative of the velogenic nature of these NDV isolates. Phylogenetic analysis showed that the new NDV strain belongs to the lineage 5d (Aldous et al., 2003) and is closely related to the Chinese strain SG/Liaoning/2009. NDV outbreaks in 2010 and 2011 have been noted in neighboring countries. Based on the high nucleotide similarity between our isolated NDV isolates and the Chinese NDV strain, the origin of these recent NDV isolates might be from China.

Survival and growth of Salmonella Typhimurium, Escherichia coli O157:H7 and Staphylococcus aureus in eggplant dip during storage

Eggplant dip is an internationally popular appetizer, prepared in some instances under uncertain hygienic conditions with inconsistent refrigeration. This study examined the effects of citric acid on the survival of pathogenic microorganisms (Salmonella Typhimurium, Escherichia coli O157:H7 and Staphylococcus aureus) and naturally present organisms (lactic acid bacteria, LAB, aerobic bacteria, APC and yeast and mold, YM) in eggplant dip during storage. Eggplant dip with 0, 0.2, 0.4, 0.6 or 0.8% citric acid was inoculated with S. Typhimurium, E. coli O157:H7 or S. aureus and stored at 4, 10 and 21 °C for ≤15 d. Throughout the study, the survival of the inoculated microorganisms was monitored, and LAB, APC, YM numbers and pH were determined. There was no significant (p>0.05) effect of citric acid on inoculated S. Typhimurium and E. coli O157:H7. Salmonella and E. coli O157:H7 survived >7d with little reduction in viability. Reduction of S. aureus viability increased with citric acid concentration and reached>3.0 log10 CFU/g by 15 d at 4 °C. Citric acid had no effect (p>0.05) on the background YM during storage at 4, 10 and 21 °C or LAB stored at 4 and 10 °C, while at 21 °C, 0.6 and 0.8% citric acid significantly reduced LAB. Citric acid had no effect (p>0.05) on the APC in samples stored at 4 °C but it had significant effects on samples stored at 10 and 21 °C. Work reported showed that the use of citric acid at 0.4-0.8% can inhibit the growth of S. aureus in eggplant dip, but adequate refrigeration is essential to minimize risk from this and other pathogens in this product.