Major difference in clinical outcome and replication of a H3N1 avian influenza strain in young pullets and adult layers.

In this study, we investigated the pathogenicity, replication and tropism of the low pathogenic avian influenza (LPAI) strain A/chicken/Belgium/460/2019(H3N1) in adult SPF layers and young SPF males. The inoculated hens showed 58% mortality and a 100% drop in egg production in the second week post inoculation. The high viral loads in the cloacal samples coincided with the period of the positive immunohistochemistry of the oviduct, acute peritonitis and time of mortality, suggesting that the replication of H3N1 in the oviduct was a major component of the onset of clinical disease and increased level of excretion of the virus. In the inoculated young birds, the clinical signs were very mild with the exception of one bird. The results suggest that the time of replication of the virus was much shorter than in the adult layers; some of the young males did not show any proof of being infected at all. To conclude, the results of the study in young birds confirmed the intravenous pathogenicity test results but also showed that the clinical signs in adult layers were very severe. Based on the mortality without a bacterial component, complete drop of egg production and post mortem findings, this H3N1 strain is a moderately virulent strain, the highest category for LPAI strains. It is important to realize that if HPAI did not exist, this moderately virulent H3N1 virus would most likely to be considered as a very virulent virus.

Induction of cystic oviducts and protection against early challenge with infectious bronchitis virus serotype D388 (genotype QX) by maternally derived antibodies and by early vaccination.

Since the end of 2003, strains of the D388 serotype (QX genotype) of infectious bronchitis virus (IBV) have caused considerable damage to the Dutch poultry industry. In order to better understand the pathogenesis of infection caused by this infectious bronchitis variant and to be able to support the poultry industry with substantiated advice to prevent or decrease the damage caused by the D388 strain, several vaccination and challenge experiments were performed in young specific pathogen free layers, young layers with maternally derived antibodies against the D388 strain and young commercial broiler breeders. The experiments confirmed the field observations that the D388 strain of the QX genotype is a very pathogenic strain that is able to cause cystic oviducts in a high percentage of birds, mortality due to nephritis and respiratory distress with complete tracheal ciliostasis and airsacculitis. Vaccination programmes using different combinations of heterologous live vaccines at day 0 or at days 0 and 14 induced a reasonable to high level of protection in the trachea, kidney, oviduct and air sacs against challenge with the D388 strain at 28 days of age. However, for very early protection, maternally-derived D388-neutralizing antibodies were shown to be very important. Titres of 9 to 10 log(2) maternally-derived D388 virus-neutralizing antibodies, which provided partial protection against tracheal damage and a high protection against replication of D388 in the kidney after challenge at 6 or 10 days of age, could be achieved using a broad heterologous live priming and subsequent boosting using inactivated IBV vaccines containing two or three heterologous IBV antigens.

Efficacy of infectious bronchitis virus vaccinations in the field: association between the alpha-IBV IgM response, protection and vaccine application parameters.

Infectious bronchitis virus (IBV) is, in spite of vaccination, still a major cause of respiratory problems in broilers and of poor egg production in breeders and layers in many parts of the world. A possible cause of the insufficient protection induced by vaccination is an inadequate application of the vaccine. This paper reports the results of two field studies. In the first, the results of the alpha-IBV IgM enzyme-linked immunosorbent assay (ELISA) on post-vaccination sera were compared with the efficacy of the IBV vaccination against homologous challenge of the same broilers. The results showed that groups with at least 50% positive sera in the IgM ELISA at 10 days post vaccination had a high level of protection against challenge. Most groups of broilers with a low level of IgM ELISA positives had a low or moderate level of protection against challenge. In a second field study, the IgM response to IBV vaccination was compared with detailed information of the vaccination process of 360 spray-vaccinated flocks of about 2-week-old broilers, layer pullets, broiler breeders and broiler grandparents. The information included parameters such as flock size, type of chicken, housing, age of the chicken, application route, vaccine, dose, water quantity and temperature, ventilation and light management, combination with other vaccines and temperature of the house. The aim was to identify factors that might be associated positively or negatively with the IgM response and thereby with the expected level of protection against homologous challenge under field conditions. Significant associations were detected between the level of IgM response and factors regarding type of bird, flock size, housing type, ventilation management, light management, age/interval of vaccination, interval between vaccination and blood sampling, and temperature of the water that was used to reconstitute the vaccine. This knowledge can be useful to improve the average efficacy of IBV vaccination in the field.

The highly pathogenic avian influenza A (H7N7) virus epidemic in The Netherlands in 2003--lessons learned from the first five outbreaks.

Clinical signs and gross lesions observed in poultry submitted for postmortem examination (PME) from the first five infected poultry flocks preceding the detection of the primary outbreak of highly pathogenic avian influenza (HPAI) of subtype H7N7 during the 2003 epidemic in the Netherlands are described. The absence of HPAI from the Netherlands for more than 75 yr created a situation in which poultry farmers and veterinary practitioners did not think of AI in the differential diagnosis as a possible cause of the clinical problems seen. Increased and progressive mortality was not reported to the governmental authorities by farmers or veterinary practitioners. It took 4 days from the first entry of postmortem material to notify the governmental authorities of a strong suspicion of an AI outbreak on the basis of a positive immunofluoresence test result. The gross lesions observed at PME did not comply with the descriptions in literature, especially the lack of hemorrhagic changes in tissues, and the lack of edema and cyanosis in comb and wattles is noted. The following lessons are learned from this epidemic: a) in the future, increased and progressive mortality should be a signal to exclude AI as cause of disease problems on poultry farms; b) intensive contact between the veterinary practitioner in the field and the veterinarian executing PME is necessary to have all relevant data and developments at one's disposal to come to a conclusive diagnosis; c) in an anamnesis, reporting of high or increased mortality should be quantified in the future (number of dead birds in relation to the number of birds brought to the farm to start production, together with the timing within the production cycle), or else this mortality cannot be interpreted properly; d) if clinical findings such as high mortality indicate the possibility of HPAI, the pathologist should submit clinical samples to the reference laboratory, even if PME gives no specific indications for HPAI; e) the best way to facilitate early detection of an HPAI outbreak is to have the poultry farmer and/or veterinary practitioner immediately report to the syndrome-reporting system currently in operation the occurrence of high mortality, a large decrease in feed or water intake, or a considerable drop in egg production; f) in order to detect low pathogenic avian influenza infections that could possibly change to HPAI, a continuous serologic monitoring system has been set up, in which commercial poultry flocks are screened for antibodies against AI virus of subtypes H5 and H7.

A cross-sectional serological survey of the Dutch commercial poultry population for the presence of low pathogenic avian influenza virus infections.

After the discovery of poultry infected with highly pathogenic avian influenza (HPAI) virus of subtype H7N7 in the central area of The Netherlands on 28 February 2003, the hypothesis was put forward that an outbreak of the low pathogenic (LP) variant of H7N7 had preceded, unnoticed, the occurrence of the HPAI virus. Consequently, a cross-sectional serological survey of the Dutch poultry population was executed in the second week of March 2003. The basic requirements set were detection of a 5% prevalence of flocks exposed to LPAI virus with 95% confidence within the production type stratification level within each province in The Netherlands. Because of supposed higher risk of avian influenza infections in ducks, turkeys and free-range poultry, all the commercial flocks of these production types present in The Netherlands were sampled. The serological screening of 28018 sera from 1193 randomly selected poultry farms, located outside surveillance zones showed that LPAI H7 virus infections had occurred on three neighbouring farms all located in the southwest of The Netherlands. No antibodies against the neuraminidase N7 subtype were detected in the sera of these farms, indicating that the subtype was different from the HPAI H7N7 subtype that caused the avian influenza epidemic in 2003. In addition, evidence of infections with non-H5 or non-H7 subtypes of influenza A virus were obtained in two other farms located in the northeast and the southeast of The Netherlands. It was concluded that the HPAI subtype H7N7 outbreak was most likely not preceded by a significant circulation of a LPAI subtype H7N7 virus. Based on the Dutch experience, recommendations are made to detect avian influenza infections faster in the future.