Daria Pestka

The prevalence and genetic characterization of Chlamydia psittaci from domestic and feral pigeons in Poland and the correlation between infection rate and incidence of pigeon circovirus

Chlamydiosis is a zoonotic disease caused by Chlamydia psittaci that occurs in a wide range of bird species. High infection rates with C. psittaci are found in pigeons, which can act as vectors transmitting this bacterium to poultry and humans. Chlamydia shedding by pigeons is intermittent and can be activated by stressors or immunosuppression. The most common immunosuppressive factor for pigeons is a pigeon circovirus (PiCV) infection. The main aim of the study was to evaluate the prevalence of C. psittaci in Polish populations of domestic and feral pigeons (Columba livia) in the context of its correlation with PiCV infections. The second objective was to determine the genetic characteristics of Polish C. psittaci isolates. The study was conducted on 377 pigeon samples (276 domestic and 101 feral pigeons) collected from pigeons from different regions of Poland. The average prevalence of C. psittaci in the Polish pigeon population was determined at 6.8%, and it was higher in domestic than in feral pigeons. This is the first ever study to suggest a potential correlation between C. psittaci and PiCV infections, which could be attributed to the fact that there are 2 to 3 times more pigeons infected with C. psittaci and coinfected with PiCV than pigeons infected with C. psittaci alone. This trend was observed mainly in the population of sick pigeons. As many as 88.2% of isolates were recognized as belonging to genotype B, and the remaining isolates were identified as belonging to genotype E. The isolates analyzed in this study demonstrated low levels of genetic variation (96-100% homology among the isolates and in relation to reference strains). Chlamydia psittaci could be expected to spread across pigeon populations due to the high probability of mutual infections between birds and the increasing number of PiCV infections.

Occurrence and genetic diversity of pigeon circovirus strains in Poland.

Pigeon circovirus (PiCV) is an immunosuppressive agent widespread throughout the world, which causes a disease in pigeons called Young Pigeon Disease Syndrome. The aim of the study was to evaluate the prevalence of PiCV in Poland and investigate the genetic diversity relative to other known PiCV isolates. Samples from 152 pigeon flocks (88 flocks of racing pigeons and 64 flocks of fancy pigeons) from various regions of Poland were tested by polymerase chain reaction and an approximately 326-base fragment of the capsid protein gene (Cap gene) of the virus was amplified. The average viral prevalence was found to be 70.3% (76.13% in racing pigeons and 62.5% in fancy pigeons). Among the obtained positive samples, 21 were selected for sequencing and a phylogenetic analysis was performed. It was found that the majority of Polish PiCV isolates, to varying degrees, are related to isolates occurring in Europe. It was also observed that the Cap gene is variable and mutations often occur in it, which impacts the amino acid sequences in the capsid protein (nucleotide similarity averaged 86.57%, amino acid similarity averaged 89.02%).

Application of pigeon circovirus recombinant capsid protein for detecting anti-PiCV antibodies in the sera of asymptomatic domestic pigeons and the potential use of a combination of serological and molecular tests for controlling circovirus infections in pigeon breeding flocks

&NA; The aim of this study was to evaluate the serologic status of domestic pigeons not infected and asymptomatically infected with the pigeon circovirus (PiCV) with the use of an enzyme‐linked assay based on PiCV recombinant capsid protein as a plate antigen. Recombinant PiCV capsid protein was produced by transforming E. coli BL21 (DE3) Rosetta colonies with expression plasmids. Blood samples and cloacal swabs were collected from 171 asymptomatic pigeons. The birds were divided into two groups (infected and not infected with PiCV) based on the results of Sybr Green real time PCR screening for the presence of PiCV genetic material. Approximately 70% of the pigeons tested positive for anti‐PiCV antibodies regardless of their infection status. Antibody levels, the coefficient of variation and standard deviation were significantly higher in the group of infected pigeons. The results indicate that ELISA is a highly useful test that complements molecular methods in evaluations of PiCV infection status in domestic pigeons. The spread of pigeon circovirus infections can be controlled by keeping breeding flocks free of PiCV, which can only be achieved by subjecting birds to real time PCR and serological tests.

Detection of Bordetella avium by TaqMan real-time PCR in tracheal swabs from wildlife birds

Bordetella avium, the causing agent of bordetellosis, a highly contagious infection of the respiratory tract in young poultry, causes significant losses in poultry farming throughout the world. Wildlife birds can be a reservoir of various pathogens that infect farm animals. For this reason the studies were conducted to estimate the prevalence of Bordetella avium in wildlife birds in Poland. Tracheal swab samples were collected from 650 birds representing 27 species. The bacterial DNA was isolated directly from the swabs and screened for Bordetella avium by TaqMan real-time PCR. The assay specificity was evaluated by testing DNA isolated from 8 other bacteria that can be present in avian respiratory tract, and there was no amplification from non-Bordetella avium agents. Test sensitivity was determined by preparing standard tenfold serial dilutions of DNA isolated from positive control. The assay revealed to be sensitive, with detection limit of approximately 4.07x10^2 copies of Bordetella avium DNA. The genetic material of Bordetella avium was found in 54.54% of common pheasants, in 9.09% of Eurasian coots, in 3.22% of black-headed gulls and in 2.77% of mallard ducks. The results of this study point to low prevalence of Bordetella avium infections in wildlife birds. The results also show that described molecular assay proved to be suitable for the rapid diagnosis of bordetellosis in the routine diagnostic laboratory.

Three-step Anti-aMPV IgA Expression Profile Evaluation in Turkeys of Different Immunological Status after TRT Vaccination.

Maternally derived antibodies (MDA) don not protect turkeys against rhinotracheitis (TRT) but high MDA influences upper respiratory tract (URT) immunity stimulation after avian Metapneumovirus (aMPV) vaccination. Humoral immunity can not be considered as an indicator of protection against TRT, but specific antibodies inhibit aMPV replication and alleviate the course of TRT. Scarce reports indicate the role of IgA in protection against TRT. The aim of our study was to investigate the impact of MDA on stimulation, antigen specificity acquisition of B lymphocytes, and the production of specific IgA after TRT vaccination of turkeys. The results of our study indicate that MDA on the day of TRT vaccination causes disturbances at different levels of specific humoral immunity expression including antigen specificity acquisition of B IgA+ lymphocytes as well as production and secretion of IgA. Vaccine immunity against aMPV associated with sIgA is well expressed in birds not possessing MDA on the day of TRT vaccination, whereas it is inhibited in MDA+ birds. These results corroborate our previous findings and indicate that MDA could be responsible for TRT vaccination failure. These findings could explain the observed frequency of TRT field outbreaks despite aMPV vaccination of turkey flocks.

Epidemiological situation of turkey coryza (bordetellosis) in Poland

The present study investigated the prevalence of Bordetella avium (BA) infections in turkey flocks of different production type in Poland, based on serological examination of blood samples collected between 2012 and 2014. The results of our study indicate that BA infections are very common in turkey flocks, they are possible during the whole production cycle and that probability of the past infection increases with age. This situation is causing high level of specific anti-BA maternally derived antibody transfer to hatching poults.

The influence of phytoncides on the immune system of broiler chickens and turkeys.

The aim of our study was to investigate the influence of adiSalmoSOL PF dietary supplement, given for 3 days in drinking water, on selected parameters of cell-mediated (Experiment I) and humoral (Experiment II) immunity in chicken and turkey broilers. In Experiment I, birds were randomly divided into two groups of 10 birds each. Group 1 comprised control turkeys or chickens, whereas group 2 birds were administered adiSalmoSOLPF. In Experiment II, a total of 69 chickens were divided into three groups (1-3) of 23 birds each. At the age of 25 days, group 1 was given adiSalmoSOLPF. Birds from groups 1-3 were vaccinated at 28 days of age. Group 2 was given adiSalmoSOLPF after vaccination. In Experiment I, a significant increase in percentages of CD4 + T lymphocytes in the bursa of Fabricius, cecal tonsils and spleen, CD8 + T lymphocytes in the thymus and spleen, CD4 + CD8 + T lymphocytes in the bursa of Fabricius and ileal mucosa and IgM + B lymphocyte in the ileal mucosa were observed in group 2 chickens. In Experiment I, a significant increase in percentages of CD4 + T lymphocytes in the thymus and spleen, CD8+ T lymphocytes in the cecal tonsils and blood, and CD4 + CD8 + T lymphocytes in the thymus and ileal mucosa was recorded in group 2 turkeys. No differences in percentages of IgM + B lymphocytes were observed between turkey groups. In Experiment II, the highest post-vaccination titers of anti-IB antibodies were observed in group 2, but it was not statistically significant. The results of our study indicate that adiSalmoSOLPF showed immunomodulatory activity in chickens and turkeys.