Infected Ducks Research Articles

PATHOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY OF P. MULTOCIDA CAPSULAR TYPE A IN TISSUES OF CHICKENS AND DUCKS INFECTED WITH FOWL CHOLERA

Sixteen chickens' flocks and six ducks᾿ flocks were surveyed for presence of fowl cholera (FC) during the period from July 2013 to November 2016. A total of 81 freshly dead birds (51 chickens of average age 50 to 180 days and 30 ducks of average age 30-100 days) were received from different farms and house breeding in Dakahlia Governorate at Mansoura Laboratory of Animal Health Research Institute for further laboratory examination. FC was diagnosed based on case history, postmortem lesions, bacteriological isolation, polymerase chain reaction (PCR), capsular typing, virulence factor detection (Tox A gene), and immunohistochemistry (IHC). In this study, Pasteurella multocida (P. multocida) were bacteriologically isolated from 22 chickens and 16 ducks. The infection occurred in 4 layer chickens flocks, 2 breeder flocks and 3 broiler flocks. Meanwhile, all affected flocks of ducks were broiler. Age susceptibility in chickens varied as the age ranged from 30-54 days in broiler flocks, 45-150 days in layer flocks during the rearing period and before start egg production, 70-150 days in breeder flocks during the rearing period. In ducks the susceptible age ranged from 30-100 days. The most affected cases of chickens were from farm housing and the most affected cases of ducks were from back yard housing. Mortality rate in chickens was 0.03 to 0.36 and in ducks was 33.3 to 46.6%. Main gross findings in affected chickens and ducks were corn meal liver, fibrinous perihepatitis and fibrinous pericarditis. Swollen hock joint and thickened foot pad were grossly detected in ducks only. The histopathological lesions in the infected ducks were more severe than those detected in the infected chickens that characterized by multiple granulomata in most examined organs. Therefore, the IHC positive reaction against antigen of P. multocida serotype A were more intensely stained and widely distributed in all examined organs of infected ducks than in chickens.

Preliminary study of the UL55 gene based on infectious Chinese virulent duck enteritis virus bacterial artificial chromosome clone

BackgroundLethal Duck Enteritis Virus (DEV) infection can cause high morbidity and mortality of many species of waterfowl within the order Anseriformes. However, little is known about the function of viral genes including the conserved UL55 gene among alpha herpes virus due to the obstacles in maintenance and manipulation of DEV genome in host cells.MethodsIn this paper, we constructed an infectious bacteria artificial chromosome (BAC) clone of the lethal clinical isolate duck enteritis virus Chinese virulent strain (DEV CHv) by inserting a transfer vector containing BAC mini-F sequence and selection marker EGFP into UL23 gene using homologous recombination. UL55 deletion and its revertant mutant were generated by two-step RED recombination in E. coli on basis of rescued recombinant virus. The function of UL55 gene in DEV replication and its effect on distribution of UL26.5 protein were carried out by growth characteristics and co-localization analysis.ResultsThe complete genome of DEV CHv can be stably maintained in E. coli as a BAC clone and reconstituted again in DEF cells. The generated UL55 deletion mutant based on DEV CHv-BAC-G displayed similar growth curves, plaque morphology and virus titer of its parental virus in infected Duck Embryo Fibroblast (DEF) cells. Immunofluorescence assay indicated that the loss of UL55 gene do not affect the distribution of UL26.5 protein in intracellular. These data also suggest infectious BAC clone of DEV CHv will facilitate the gene function studies of DEV genome.ConclusionsWe have successfully developed an infectious BAC clone of lethal clinical isolate DEV CHv for the first time. The generated UL55 gene mutant based on that demonstrated this platform would be a very useful tool for functional study of DEV genes. We found the least known DEV UL55 is dispensable for virus replication and UL26.5 distribution, and it could be a very promise candidate locus for developing bivalent vaccine. Experiment are now in progress for testifying the possibility of UL55 gene locus as an exogenous gene insertion site for developing DEV vectored vaccine.

Disruption of the M949_RS01915 gene changed the bacterial lipopolysaccharide pattern, pathogenicity and gene expression of Riemerella anatipestifer

Riemerella anatipestifer is an important pathogen that causes septicemia anserum exsudativa in ducks. Lipopolysaccharide (LPS) is considered to be a major virulence factor of R. anatipestifer. To identify genes involved in LPS biosynthesis, we screened a library of random Tn4351 transposon mutants using a monoclonal antibody against R. anatipestifer serotype 1 LPS (anti-LPS MAb). A mutant strain RA1067 which lost the reactivity in an indirect ELISA was obtained. Southern blot and sequencing analyses indicated a single Tn4351 was inserted at 116 bp in the M949_RS01915 gene in the RA1067 chromosomal DNA. Silver staining and Western blot analyses indicated that the RA1067 LPS was defected compared to the wild-type strain CH3 LPS. The RA1067 displayed a significant decreased growth rate at the late stage of growth in TSB in comparison with CH3. In addition, RA1067 showed higher susceptibility to complement-dependent killing, more than 360-fold attenuated virulence based on the median lethal dose determination, increased bacterial adhesion and invasion capacities to Vero cells and significantly decreased blood bacterial loads in RA1067 infected ducks, when compared to the CH3. An animal experiment indicated that inactivated RA1067 cells was effective in cross-protecting of the ducks from challenging with R. anatipestifer strains WJ4 (serotype 1), Yb2 (serotype 2) and HXb2 (serotype 10), further confirming the alteration of the RA1067 antigenicity. Moreover, RNA-Seq analysis and real-time PCR verified two up-regulated and three down-regulated genes in RA1067. Our findings demonstrate that the M949_RS01915 gene is associated to bacterial antigenicity, pathogenicity and gene regulation of R. anatipestifer.

Identification of the ferric iron utilization gene B739_1208 and its role in the virulence of R. anatipestifer CH-1

Riemerella anatipestifer is an important bacterial pathogen in ducks and causes heavy economic losses in the duck industry. However, the pathogensis of this bacterium is poorly understood. In this study, a putative outer membrane hemin receptor gene B739_1208 in R. anatipestifer CH-1 was deleted to determine the relationship between iron uptake and virulence. The R. anatipestifer CH-1ΔB739_1208 mutants grew significantly more slowly than the wild-type bacteria in TSB liquid medium. Further characterization revealed that the R. anatipestifer CH-1ΔB739_1208 mutants were deficient in iron uptake. Animal experiments indicated that the median lethal dose of the wild-type RA-CH-1 in ducklings was 3.89×108, whereas the median lethal dose of the R. anatipestifer CH-1ΔB739_1208 mutant in ducklings was 5.68×109. The median lethal dose of the complementation strain in ducklings was 9.84×108. Additional analysis indicated that bacterial loads in the blood, liver, and brain tissues in the R. anatipestifer CH-1ΔB739_1208-infected ducklings were significantly decreased compared to those in the wild-type R. anatipestifer CH-1 infected ducklings. In a duck co-infection model with R. anatipestifer CH-1 and R. anatipestifer CH-1ΔB739_1208, the R. anatipestifer CH-1B739_1208 mutant was outcompeted by the wild-type R. anatipestifer CH-1 in the blood (P<0.002), livers (P<0.001) and brains (P<0.001) of infected ducks, indicating that B739_1208 gene expression provided a competitive advantage in these organs. Our results demonstrate that the B739_1208 gene is a virulence factor in R. anatipestifer CH-1.

Identification of Ribonuclease Z Gene from an Outbreak of Riemerella anatipestifer Infection in Ducks of Bangladesh

Duck septicemia is a deadly disease of duck causing huge economic losses in duck rearing areas. Despite its importance, no study was devoted to unravel the Riemerella (Pasteurella) anatepestifer in Bangladesh. Therefore, a systemic investigation was carried out to identify the isolate targeting ribonuclease Z gene from an outbreak in 2013-2014. Samples were collected from sixty randomly selected dead ducks from three outbreaks areas of North East Bangladesh (Netrokona district). Histopathological, cultural and sugar fermentation test of all samples revealed the infectivity with R. anatipestifer. Polymerase chain reaction specific to R. anatipestifer identified the 421 bp fragment of ribonuclease Z gene in thirty seven cases. Identified gene of the isolate was sequenced (Accession No. KU992381) and phylogenetic lineage was analyzed for the first time in Bangladesh. The phylogenetic analysis showed that Bangladeshi isolate R. anatipestifer BD 2014 resembles highest similarity with the R. anatipesrifer isolated from ducks of china. This study provide evidence that duck septicemia could be the main culprit of mortality in growing ducks in the duck rearing areas. Further investigation is needed to identify secondary or co-infection state as well as the prevalence of this infection including the economic burdens in duck industry in Bangladesh.

Experimental infection of mandarin duck with highly pathogenic avian influenza A (H5N8 and H5N1) viruses

A highly pathogenic avian influenza (HPAI) H5N8 virus was first detected in poultry and wild birds in South Korea in January 2014. Here, we determined the pathogenicity and transmissibility of three different clades of H5 viruses in mandarin ducks to examine the potential for wild bird infection. H5N8 (clade 2.3.4.4) replicated more efficiently in the upper and lower respiratory tract of mandarin ducks than two previously identified H5N1 virus clades (clades 2.2 and 2.3.2.1). However, none of the mandarin ducks infected with H5N8 and H5N1 viruses showed severe clinical signs or mortality, and gross lesions were only observed in a few tissues. Viral replication and shedding were greater in H5N8-infected ducks than in H5N1-infected ducks. Recovery of all viruses from control duck in contact with infected ducks indicated that the highly pathogenic H5 viruses spread horizontally through contact. Taken together, these results suggest that H5N8 viruses spread efficiently in mandarin ducks. Further studies of pathogenicity in wild birds are required to examine possible long-distance dissemination via migration routes.

Characterizations of H4 avian influenza viruses isolated from ducks in live poultry markets and farm in Shanghai.

H4 avian influenza virus is one of the most prevalent influenza virus subtypes in birds. The evolution and pathogenicity of H4 AIV in domestic birds of China remain largely unclear. In the present study, a total of eight H4 AIV strains isolated in duck farm and live poultry markets (LPM) were characterized. Phylogenetic analysis indicated that these strains are divided into two groups in the Eurasian lineage. Eight genes of MH-2/H4N6 isolated from a duck farm were closely related to three H4N6 viruses from LPM, suggesting a potential AIV link between farms and LPMs. Additionally, the HA, NA, PB2, NP, and NS genes of two other H4N6 viruses isolated in LPM clustered with that of MH-2/H4N6. However, the remaining genes were more closely related to other sublineages, suggesting that MH-2/H4N6-originated viruses reassorted with other viruses in LPM. All H4 viruses replicated in mouse lungs without prior adaptation and all viruses replicated and transmitted among ducks. 29-1/H4N2, MH-2/H4N6, and 420-2/H4N6 viruses caused systemic infection in infected ducks. However, most of the viruses were not adapted in chickens. The present results indicate a potential correlation of AIV between LPMs and farms and suggest that active surveillance of AIV in LPM is warranted in China.

Seroepidemiological Evidence for the Presence of Japanese Encephalitis Virus Infection in Ducks, Chickens, and Pigs, Bali-Indonesia

Background: The presence of various animals, such as: ducks, chickens and pigs in households increases the potential risks of zoonosis from animal to human. One of the diseases is Japanese encephalitis (JE). The seroepidemiological studies on the presence JE among animals especially those raised in household is very important for emerging and reemerging disease control program. Ducks, chickens and pigs have long been considered as carrier and even the amplifier hosts of Japanese encephalitis virus (JEV) replication. The presence of the animal hosts and mosquitoes as vector could result in transmission of the JEV to humans. Methods: A seroepidemiological study of the presence of Japanese encephalitis virus (JEV) was conducted by collecting sera and detecting the antibody against JEV in ducks, chickens and pigs in Bali. As pig is the amplifying animal of JEV, comparison JEV antibody between ducks reared in households with pig nearby and with no pig were also determined the presence of antibody against JEV was examined by using indirect enzyme-linked immunosorbent assay (ELISA). The serum samples with over cut off value (COV) of optical density were considered as those containing Ab against JEV. Results: Antibody against JEV was demonstrated in ducks (20.6%), chickens (36.7%) and pigs (32.2%) evaluated in this study. Moreover, there was no significant difference (p&gt;0.05) in the prevalence of antibody against JEV in ducks kept closely with pigs compared to the antibody in the ducks reared without pigs around. Conclusion: This study convinced that antibody against JEV is found in ducks, chickens and pigs in Bali. Indicating that these animals was infected or previously infected by the virus.

Construction of a recombinant duck enteritis virus vaccine expressing hemagglutinin of H9N2 avian influenza virus and evaluation of its efficacy in ducks.

H9 subtype avian influenza viruses (AIVs) remain a significant burden in the poultry industry and are considered to be one of the most likely causes of any new influenza pandemic in humans. As ducks play an important role in the maintenance of H9 viruses in nature, successful control of the spread of H9 AIVs in ducks will have significant beneficial effects on public health. Duck enteritis virus (DEV) may be a promising candidate viral vector for aquatic poultry vaccination. In this study, we constructed a recombinant DEV, rDEV-∆UL2-HA, inserting the hemagglutinin (HA) gene from duck-origin H9N2 AIV into the UL2 gene by homologous recombination. One-step growth analyses showed that the HA gene insertion had no effect on viral replication and suggested that the UL2 gene was nonessential for virus growth in vitro. In vivo tests further showed that the insertion of the HA gene in place of the UL2 gene did not affect the immunogenicity of the virus. Moreover, a single dose of 103 TCID50 of rDEV-∆UL2-HA induced solid protection against lethal DEV challenge and completely prevented H9N2 AIV viral shedding. To our knowledge, this is the first report of a DEV-vectored vaccine providing robust protection against both DEV and H9N2 AIV virus infections in ducks.

Different Duck Species Infected Intramuscularly with Duck-Origin Genotype IX APMV-1 Show Discrepant Mortality and Indicate Another Fatal Genotype APMV-1 to Ducks

Isolations of genotype IX (gIX) avian paramyxovirus type 1 (APMV-1) from various bird species have been more common recently, with isolates showing variable pathogenicity in different species of poultry. Here we sequenced the genome of a Muscovy duck origin gIX virus strain XBT14 and characterized the virulence and pathogenicity of this isolate in chickens and ducks. The genome sequence of strain XBT14 is 15,192 nt in length, containing multiple basic amino acids at the fusion protein cleavage site. The XBT14 strain shared 91.6%-91.9% nucleotide identities with early-genotype viruses (such as genotype III and IV) and shared 85.3%-85.9% nucleotide homologies with later genotype viruses (such as genotype VII). Pathogenicity tests showed that strain XBT14 could cause death in different duck breeds with a mortality rate of 44.4% in Muscovy duck, 25.9% in Sheldrake, and 11.1% in Cherry Valley duck, respectively. Similar mortality discrepancies were also observed in different ducks when infected with chicken-origin gIX virus strain F48E8. These results indicate that XBT14-like velogenic gIX APMV-1 (such as XBT14, F48E8, and GD09-2) could cause fatal infection in duck, and genotype IX is another genotype velogenic to duck as well as genotype VII. Accompanied by genetic differences in the vaccine strains or dominant strains prevailing in poultry, the virulent XBT14-like gIX viruses might become potentially endemic strains in poultry in the future.

Hepatitis Virus Infections in Poultry.

Viral hepatitis in poultry is a complex disease syndrome caused by several viruses belonging to different families including avian hepatitis E virus (HEV), duck hepatitis B virus (DHBV), duck hepatitis A virus (DHAV-1, -2, -3), duck hepatitis virus Types 2 and 3, fowl adenoviruses (FAdV), and turkey hepatitis virus (THV). While these hepatitis viruses share the same target organ, the liver, they each possess unique clinical and biological features. In this article, we aim to review the common and unique features of major poultry hepatitis viruses in an effort to identify the knowledge gaps and aid the prevention and control of poultry viral hepatitis. Avian HEV is an Orthohepevirus B in the family Hepeviridae that naturally infects chickens and consists of three distinct genotypes worldwide. Avian HEV is associated with hepatitis-splenomegaly syndrome or big liver and spleen disease in chickens, although the majority of the infected birds are subclinical. Avihepadnaviruses in the family of Hepadnaviridae have been isolated from ducks, snow geese, white storks, grey herons, cranes, and parrots. DHBV evolved with the host as a noncytopathic form without clinical signs and rarely progressed to chronicity. The outcome for DHBV infection varies by the host's ability to elicit an immune response and is dose and age dependent in ducks, thus mimicking the pathogenesis of human hepatitis B virus (HBV) infections and providing an excellent animal model for human HBV. DHAV is a picornavirus that causes a highly contagious virus infection in ducks with up to 100% flock mortality in ducklings under 6 wk of age, while older birds remain unaffected. The high morbidity and mortality has an economic impact on intensive duck production farming. Duck hepatitis virus Types 2 and 3 are astroviruses in the family of Astroviridae with similarity phylogenetically to turkey astroviruses, implicating the potential for cross-species infections between strains. Duck astrovirus (DAstV) causes acute, fatal infections in ducklings with a rapid decline within 1-2 hr and clinical and pathologic signs virtually indistinguishable from DHAV. DAstV-1 has only been recognized in the United Kingdom and recently in China, while DAstV-2 has been reported in ducks in the United States. FAdV, the causative agent of inclusion body hepatitis, is a Group I avian adenovirus in the genus Aviadenovirus. The affected birds have a swollen, friable, and discolored liver, sometimes with necrotic or hemorrhagic foci. Histologic lesions include multifocal necrosis of hepatocytes and acute hepatitis with intranuclear inclusion bodies in the nuclei of the hepatocytes. THV is a picornavirus that is likely the causative agent of turkey viral hepatitis. Currently there are more questions than answers about THV, and the pathogenesis and clinical impacts remain largely unknown. Future research in viral hepatic diseases of poultry is warranted to develop specific diagnostic assays, identify suitable cell culture systems for virus propagation, and develop effective vaccines.

Riemerella anatipestifer M949_1360 Gene Functions on the Lipopolysaccharide Biosynthesis and Bacterial Virulence

Riemerella anatipestifer causes septicemic and exudative diseases in poultry, resulting in major economic losses to the duck industry. Lipopolysaccharide (LPS), as an important virulence factor in Gram-negative bacteria, can be recognized by the immune system and plays a crucial role in many interactions between bacteria and animal hosts. In this study, we screened out one LPS defective mutant strain RAΔ604 from a random transposon mutant library of R. anatipestifer serotype 1 strain CH3, which did not react with the anti-CH3 LPS monoclonal antibody 1C1 in an indirect enzyme-linked immunosorbent assay. Southern blot analysis confirmed that the genome of RAΔ604 contained a single Tn4351 insert. Then, we found that the M949_1360 gene was inactivated by insertion of the transposon. Using silver staining and western blot analyses, we found that the LPS pattern of RAΔ604 was defective, as compared with that of the wild-type (WT) strain CH3. The mutant strain RAΔ604 showed no significant influence on bacterial growth, while bacterial counting and Live/dead BacLight Bacterial Viability staining revealed that bacterial viability was decreased, as compared with the WT strain CH3. In addition, the abilities of the mutant strain RAΔ604 to adhere and invade Vero cells were significantly decreased. Animal studies revealed that the virulence of the mutant strain RAΔ604 was decreased by more than 200-fold in a duck infection model, as compared with the WT strain CH3. Furthermore, immunization with live bacteria of the mutant strain RAΔ604 protected 87.5% ducks from challenge with R. anatipestifer serotype 1 strain WJ4, indicating that the mutant strain RAΔ604 could be used as a potential vaccine candidate in the future.

Increased efficacy of inactivated vaccine candidates prepared with Salmonella enterica serovar Typhimurium strains of predominant genotypes in ducks

Salmonella enterica serovar Typhimurium has been a major causative agent of food-borne human disease, mainly due to consumption of contaminated food animal products. In particular, ducks serve as a reservoir of serovar Typhimurium, and are one of the common sources of human infection. To prevent infection of ducks, and therefore minimize human infection, it is critical to control the persistent epidemic strains in ducks. Here, we analyzed the genetic diversity and virulence of serovar Typhimurium isolates from ducks in Korea to identify the predominant strains that might be used as efficient vaccine candidates for ducks. Among the isolates, 2 representative isolates (ST26 and ST76) of predominant genotypes were selected as vaccine strains on the basis of genotypic analysis by pulsed-field gel electrophoresis and DNA microarrays. Two-week-old ducks were then injected intramuscularly with inactivated vaccine candidates prepared using ST26 or ST76 (10(8) cfu/0.5 mL/duck or 10(9) cfu/0.5 mL/duck), and oral challenge with a highly virulent serovar Typhimurium strain (10(9) cfu/0.5 mL/duck) was carried out 2 wk later. Shedding of the challenge strain was significantly decreased in group 2 after vaccination. The antibody levels by enzyme-linked immunosorbent assay in all vaccinated groups were enhanced significantly (P < 0.05) compared to the unvaccinated control group. Overall, vaccination with ST26 or ST76 reduced bacterial shedding and colonization in internal organs, and induced elevated antibody response. In particular, serovar Typhimurium ST26 (10(8) cfu/0.5 mL/duck) was the most effective vaccine candidate, which can provide efficient protection against serovar Typhimurium in ducks with higher effectiveness compared to a commercial vaccine currently used worldwide.

The pathogenicity of novel duck reovirus in Cherry Valley ducks

The novel duck reovirus (NDRV) is an emerging, contagious infection. To better realize the pathogenic mechanism of NDRV in ducks, an infection experiment was conducted. The resulting data demonstrated that typical gross lesions were observed in the infected ducks. NDRV was able to replicate in various tissues, leading to these pathological lesions, especially on the liver and spleen. Real-time quantitative PCR showed that the expression of most innate immune-related genes was up-regulated and the antiviral innate immune response could be established in both the liver and spleen. This study indicates that NDRV is a pantropic virus. To resist viral infection, several pathogen recognition receptors can cooperatively recognize NDRV and initiate innate immunity, but the responses are different between different tissues. As far as we know, this is the first systematic investigation of the pathogenicity of NDRV in Cherry Valley ducks based on the host’s innate immunity, and these data will provide new insights into the further study of the disease.

The effect of Tembusu virus infection in different week-old Cherry Valley breeding ducks

To study the effect of Tembusu virus (TMUV) infection on Cherry Valley Breeding ducks of different ages, 350 five-week-old ducks were divided into 14 groups. Ducks in seven experimental group were respectively infected with 1.265×105 mean embryo lethal dose (ELD50) of TMUV-AHQY strain (in 4.2mL) by intravenous route. Ducks in control groups were inoculated with Phosphate-buffered Saline (PBS) in the same way. Clinical symptoms, gross and microscopic lesions, viral loads and serum antibodies were detected and recorded for 20days after infection. Some ducks infected at 7 and 21 week s of age showed severe clinical symptoms including depression and inappetence, and no obvious clinical symptoms were seen in other week-old infected ducks. Severe gross lesions including hepatomegaly, meningeal congestion, myocardial hemorrhage, intestinal, myocardial and pulmonary edema were observed in ducks infected at 7, 18 and 21 weeks of age. No or mild gross lesions were observed in ducks infected at 14 and 16 weeks of age. The main microscopic lesions including hyperaemia, degeneration and necrosis of different cells and inflammatory cellular infiltration mainly consisting of mononuclear cells or lymphocytes were observed in ducks infected at 7 and 21 week of age. But relatively intact structures and rare lymphocytic infiltration were presented in ducks infected at 14 and 16 weeks of age. Viral antigen was more frequently observed in organ slices collected from 7 week-old infected ducks and few positive staining was found in 14 and 16 week-old infected ducks. Less viral loads in different tissues and swabs were detected by a quantitative real-time PCR assay. The level of viral loads in the tissues of ducks infected at 14 and 16 weeks of age was very lower than that of ducks infected at 7 and 21 weeks of age. Meanwhile, less viral copy numbers were detected in swab samples collected from 14 and 16 week-old infected ducks. Ducks infected at 14-week-old developed significantly higher serum neutralizing antibody titers than those infected at other week of age. These results indicated that the effect of TMUV infection on Cherry Valley ducks is partly related to weeks of age. 7–10 week-old and 18–21 week-old ducks were more susceptible to TMUV infection, but 14–16 week-old ducks were more resistant to this disease.

Genome analysis of a low-pathogenic H5N2 influenza virus that kills chicken embryos

Veterinaria México OA ISSN: 2448-6760Cite this as:Steffani Hernández G, Chávez Maya F, Rojas Anaya E, Loza Rubio E, García Espinosa G. Genomic analysis of an atypical Mexican low-pathogenic H5N2 avian influenza virus. Veterinaria México OA. 2016;3(2). doi: 10.21753/vmoa.3.2.363We analysed the genome of a low-pathogenic avian H5N2 influenza virus isolated from the faeces of experimentally infected Pekin ducks and Leghorn-type chickens to determine its origin and molecular characteristics. The complete genomic sequence was determined using a Sanger-based genome sequencing method and was subsequently characterized by phylogenetic analysis and genetic comparison. The results of this study showed that 8 genomic segments corresponded to an avian influenza virus that were related with strains isolated in Mexico. Investigation of the haemagglutinin gene revealed the presence of few basic amino acids at the cleavage site and lack of a potential N-glycosylation site at position 11. The gene encoding the PB1 protein lacked PB1-F2 and the basic polymerase gene codes for PA-X. In addition, the basic polymerase gene contained the consensus ribosomal frameshifting motif TCC TTT CGT C, which is required for the expression of the PA-X. Molecular characteristics showed that the virus has features of a low-pathogenic H5 influenza virus with the exception of a potential N-glycosylation site at position 11. The genome information for this particular virus will provide a molecular map for further in vivo studies to identify why some influenza viruses can persist in chickens for long periods of time. Such information will be useful in countries such as Mexico, where the virus has been a poultry health problem since 1994 and has the potential to evolve high pathogenicity.Figure 1. Phylogenetic tree of the HA gene of A/chicken/Mexico/2007 (H5N2) Influenza A virus []. The phylogenetic tree was constructed using the neighbour-joining method. The percentages of replicate trees in which the associated taxa clustered together in a bootstrap test (1,000 replicates) are shown next to the branches. The evolutionary distances were computed using the Kimura 2-parameter method. The analysis involved 101 chicken nucleotide sequences. Evolutionary analyses were conducted in MEGA 5.05. A larger triangle size represents a larger number of nucleotide sequences with genetic relationship.

Genomic analysis of an atypical Mexican low-pathogenic H5N2 avian influenza virus

We analysed the genome of a low-pathogenic avian H5N2 influenza virus isolated from the faeces of experimentally infected Pekin ducks and Leghorn-type chickens to determine its origin and molecular characteristics. The complete genomic sequence was determined using a Sanger-based genome sequencing method and was subsequently characterized by phylogenetic analysis and genetic comparison. The results of this study showed that 8 genomic segments corresponded to an avian influenza virus that were related with strains isolated in Mexico. Investigation of the haemagglutinin gene revealed the presence of few basic amino acids at the cleavage site and lack of a potential N-glycosylation site at position 11. The gene encoding the PB1 protein lacked PB1-F2 and the basic polymerase gene codes for PA-X. In addition, the basic polymerase gene contained the consensus ribosomal frameshifting motif TCC TTT CGT C, which is required for the expression of the PA-X. Molecular characteristics showed that the virus has features of a low-pathogenic H5 influenza virus with the exception of a potential N-glycosylation site at position 11. The genome information for this particular virus will provide a molecular map for further in vivo studies to identify why some influenza viruses can persist in chickens for long periods of time. Such information will be useful in countries such as Mexico, where the virus has been a poultry health problem since 1994 and has the potential to evolve high pathogenicity.

Prevalence of Opisthorchis viverrini-Like Fluke Infection in Ducks in Binh Dinh Province, Central Vietnam.

Following the first report of Opisthorchis viverrini infection in a domestic duck in Phu My District of Binh Dinh Province, Central Vietnam, many other cases were observed in the province. We determined the infection rate and intensity of O. viverrini infection in ducks in 4 districts of the province. A total of 178 ducks were randomly selected from 34 farms for examination of flukes in the liver and gall bladder. An infection rate of 34.3% (range 20.7-40.4% among districts) was found; the intensity of infection was 13.8 worms per infected duck (range 1-100). These findings show the role of ducks as a host for O. viverrini, duck genotype, which is sympatric with the human O. viverrini genotype in this province. It also stresses the need for investigations on the zoonotic potential and the life cycle of this parasite.

Upregulation of duck interleukin-17A during Riemerella anatipestifer infection

Although IL-17 cytokines play critical roles in host defense immunity, dysregulated expression of these cytokines is associated with inflammation and autoimmune diseases. Riemerella anatipestifer is the most important infectious bacterium in the duck industry. Interestingly, not all avian species are equally susceptible to R. anatipestifer infection. This paper reports the first description of mortality rate, bacterial burden, and expression profiles of immune-related genes between ducks and chickens infected with R. anatipestifer. Ducks exhibited increased susceptibility to R. anatipestifer infection compared to chickens, as determined by mortality rate and bacterial burden. Comparative expression analyses of immune-related genes in R. anatipestifer-infected tissues obtained from both species revealed that TLR3, TLR7, IL-2, IL-4, and IFN-γ transcript levels were higher in chickens, whereas TLR4 and IL-17A transcript levels were higher in ducks. Marked increases in expression of IL-17A and IL-6, but not TGF-β, were associated with Th17 cell differentiation in duck splenic lymphocytes, but not in chicken splenic lymphocytes, stimulated with R. anatipestifer. Moreover, upregulation of IL-1β, IL-6, and IL-17A mRNA expressions, but not TGF-β, was confirmed in the liver and spleen of ducks infected with R. anatipestifer, indicating that IL-17A is strongly associated with Riemerella infection in ducks.

Genome Sequence of Riemerella anatipestifer Strain RCAD0122, a Multidrug-Resistant Isolate from Ducks

Riemerella anatipestifer is an important pathogenic bacterium in waterfowl and other avian species. We present here the genome sequence of R. anatipestifer RCAD0122, a multidrug-resistant strain isolated from infected ducks. The isolate contains at least nine types of antibiotic resistance-associated genes.