Four years of African swine fever in Poland. New insights into epidemiology and prognosis of future disease spread.

Four and a half years of African Swine Fever (ASF) in population of free-ranging wild boars and domestic pigs revealed a number of novel insights into the disease epidemiology. Until No- vember 20th, 2018, in total 3048 cases in wild boars and 213 outbreaks in domestic pigs have been confirmed. In spite of low contagiosity as well as low rate of ASF spread in wild boars the disease has an enormous socio-economical impact on the production of pigs in Poland. One of the most important aspects which directly influences the dynamics of ASF spread is the unpredictable hu- man activity. Another important factor responsible for continuous ASF spread is fast recovery of wild boar population in spite of efforts taken by hunters. Assuming our scientific opinion ASF seems to be present in wildlife for the incoming few or several years. Therefore, extraordinary measures should be prepared and undertaken to limit the risk of the occurrence of future out- breaks in domestic pigs. One of the most crucial issues is implementation of strict biosecurity measures in all domestic pigs holdings.

Seroprevalence of Coxiella burnetii among domestic ruminants and horses in Poland.

Coxiella burnetii is the causative agent of Q fever. The cases of the disease are recorded in various species, including domestic animals. The aim of this investigation was to estimate the seroprevalence of C. burnetii in ruminants including cattle, sheep, goats, and horses. Totally, 2082 serum samples from 936 goats, 933 cattle, 89 sheep, and 124 horses, including various horse breeds, were tested by ELISA or complement fixation test. The examination revealed that Polish horses are seronegative while in the populations of cattle and small ruminants, seropositive animals are presented. The percentage of seropositive cattle, goats and sheep was 4.18, 6.30, and 13.48, respectively.

Development of cross-priming amplification for direct detection of the African Swine Fever Virus, in pig and wild boar blood and sera samples.

UNLABELLED: African swine fever (ASF) is considered a major threat to the production of pigs worldwide. The ASF aetiological agent, ASFV, is the sole member of the Asfivirus genus, belonging to the Asfarviridae family. An effective ASF vaccine is not currently available, thus the only measures of ASF spread control include, reliable and fast diagnosis. Officially approved, diagnostic methods include, virus isolation, serological assays, including enzyme-linked immunosorbent assay and immunoperoxidase assay (IPT) and different modifications of the polymerase chain reaction (PCR). This paper describes the first development and application of a cross-priming amplification method (CPA) for the direct detection of genetic ASFV material, in blood and sera from pigs and wild boars. This method is specific only to ASFV DNA. The study showed that CPA had equal sensitivity, in comparison to the official, universal probe library (UPL) real-time PCR and reached 7·2 copies of standard plasmid DNA, containing a p72 gene fragment. This method was capable of detecting ASFV DNA in all examined blood samples, originating from pigs; n = 10 and wild boars; n = 76. The obtained results were also confirmed by the officially approved, real-time PCR. The developed CPA might be further used by local and county veterinary officers, hunters or pig farmers, for preliminary ASF diagnosis. SIGNIFICANCE AND IMPACT OF THE STUDY: The spread of the African swine fever virus (ASFV) among infected pigs and wild boars, is currently one of the most important facets of virus transmission in eastern Europe. Cross-priming amplification (CPA) has been developed, for fast and direct development of genetic ASFV material in the blood and sera of infected pigs and wild boars. It has been shown that CPA is a rapid, sensitive and specific isothermal method for the detection of ASFV DNA, in directly collected blood or sera from pigs and wild boars.

Cytometric analysis of surface molecules of leucocytes and phagocytic activity of granulocytes and monocytes/macrophages in cows with pyometra.

Pyometra is a serious problem in dairy cow herds, causing large economic losses due to infertility. The development of pyometra depends mainly on the immunological status of the cow. The aim of the study was a comparative evaluation of selected indicators involving non-specific and specific immunity in cows with pyometra and in cows without inflammation of the uterus. The study was performed in 20 cows, which were divided into two groups: pyometra group and healthy group, each comprising 10 cows, based on the results of cytological and ultrasonographic tests. A flow cytometric analysis was performed for the surface molecules CD4, CD8, CD14, CD21, CD25 and CD4(+) CD25(+) on leucocytes, and the phagocytic activity was determined from granulocytes and monocytes/macrophages in the peripheral blood and uterine washings, respectively. It was demonstrated that the percentage of phagocytic granulocytes and monocytes/macrophages in both the peripheral blood and uterine washings was significantly lower in cows with pyometra compared with the healthy group (p < 0.001). Significantly (p ≤ 0.001) lower percentage of CD4(+) , CD14(+) , CD25(+) and CD4(+) CD25(+) phenotype leucocytes was also observed in the peripheral blood of cows from the pyometra group, along with a significantly higher (p < 0.001) percentage of CD8(+) and CD21(+) lymphocytes as compared to the healthy group. The results of work indicate that disfunction of cell immunity coexisting with pyometra may be caused by a bacterial infection and the presence of blocking agents (IL-10), released by the increasing number of CD8(+) lymphocytes what leads to the advanced inflammation of uterus.

Epidemiology of African Swine Fever in Poland since the detection of the first case.

The purpose of this paper is to provide characteristics of the spread of African Swine Fever (ASF) in Poland from February to August, 2014. The samples from dead wild boar and domestic pigs were submitted to the National Veterinary Research Institute, National Reference Laboratory for ASF in Pulawy, Poland, for testing by PCR and ELISA methods. In the studied period, fourteen cases of ASF in wild boar and two outbreaks in backyard pigs were confirmed. In addition to the results of laboratory tests performed in 2014, the article describes the ASF surveillance programme in wild boar and pigs in Poland carried out in 2011-2013. The spread of ASF in Poland is compared with the epidemiological situation in Lithuania, Latvia, Belarus and the Russian Federation.

Rapid detection of Chlamydia/Chlamydophila group in samples collected from swine herds with and without reproductive disorders.

The study was carried out in seven reproductive herds of pigs. In three of them reproductive disorders were observed. Three herds consisted of 10-50 and four consisted of 120-500 adult sows and they were called small and medium, respectively. Fifty-seven adult sows were randomly selected from herds. Serum samples were tested using the complement fixation test and swabs from both eyes and from the vaginal vestibule were examined using real-time PCR. All serum samples were negative. Infected sows were present in each of the study herds. In total, there were 28 positive samples (53%, 28/48) in real-time PCR in sows with reproductive disorders and 35 (53%, 35/66) in sows selected from herds without problems in reproduction. One isolate proved to be Chlamydophila pecorum, whereas all the remaining were Chamydia suis.

Changes in the peripheral leukocyte phenotype of calves in clinical cases of bronchopneumonia complicated with chlamydial co-infectious agent.

This paper reports the alterations in peripheral blood leukocyte phenotype in respiratory diseased calves affected with chlamydial and non-chlamydial co-infectious agent. The etiological contribution of chlamydial infectious agent in examined clinical cases of enzootic bronchopneumoniae syndrome was confirmed in affected calves serologically both by complement fixation test (CF) and enzyme immunoassay (EIA). Changes in leukocyte subpopulations in the blood of the calves were detected both with routine haematological methods and by FCM using specific monoclonal antibodies directed against CD14, CD45, CD2, CD4, CD8 and WC4 (a specific surface marker for bovine B-lymphocytes). The results obtained by flow cytometry analysis indicate that polymorfonuclear neutrophils (PMNLs) and T lymphocytes, especially CD8-positive cells, may play a significant role in cellular immune response against Chlamydophila psittaci (Chl. psittaci) co-infection in calves suffering from enzootic bronchopneumonia syndrome. A repercussion of this was a significant increase of the cell numbers in peripheral blood of the infected animals. Effective recruitment from a reserve marginal pool of these cells into blood vessels and activation of bone marrow proliferation are probably the reason for their high circulating number.

Pathogenicity of the microorganisms of the family Chlamydiaceae respecting the changes in their classification.

The comparison of names of the species before the reclassification of the family Chlamydiaceae into two genera with the species names acknowledged at present is presented. The pathogenicity of these species for animals and man is characterised.