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Infectious bursal disease virus is an important poultry pathogen. It is distributed worldwide and causes significant economic losses. In this study, a system was adopted for the simultaneous monitoring of vaccine and virulent strains using reverse transcription polymerase chain reaction (RT-PCR). After the decay of maternal antibodies, chickens were vaccinated at the age of 37 days with a virus of intermediate virulence and challenged at 5, 10 and 14 days post vaccination (dpv). The challenge was done with IBDV strain CH/99. Sequencing of the hypervariable region of VP2 has shown that CH/99 belongs to the very virulent group of viruses. The vaccine virus could be found in the bursa of Fabricius, spleen, thymus and bone marrow until 24 dpv. The CH/99 challenge virus was found in the bursa and lymphoid organs when chickens were challenged at 5 and 10 dpv. When challenge was performed at 14 dpv, the pathogenic virus could not be found in the bursa and other lymphoid organs.
Partial genome sequence of a herpes-like virus, isolated from Siberian sturgeon ( Acipenser baeri ), was determined and subjected to phylogenetic analysis. The virus (SbSHV) has been shown to be the causative agent of an acute disease with high mortality in farmed juvenile sturgeons in Russia. Two fragments (of 7000 and 300 base pairs in length) encompassing 3 complete and 3 partial ORFs were amplified by PCR. Sturgeon herpesvirus strains, classified into species Acipenserid herpesvirus 2 (AciHV-2), have been isolated and partially sequenced from several regions (California, Idaho, Oregon and Canada) of North America from white ( A. transmontanus) and shortnose sturgeons ( A. brevirostrum ). The sequence of the SbSHV strain shared highest identity with that of the Canadian strain originating from shortnose sturgeon. The phylogenetic analysis also confirmed that SbSHV is closely related to AciHV-2 and could also be classified into this virus species. This is the first report on the occurrence of AciHV-2 in Europe. Previously, only another virus species, AciHV-1 has been detected in farmed white sturgeons in Italy. The size and position of ORFs in the examined gene block confirmed that this genomic region is highly conserved in members of the genus Ictalurivirus .
A survey was conducted to evaluate the presence and prevalence of Porcine Bocavirus (PBoV) in Croatian domestic pigs by means of PCR targeting the NS1 gene fragment of PBoV. This study included testing of faecal samples collected from 10 small commercial farms and 11 small backyard holdings in Croatia. The presence of PBoV was confirmed by PCR in 24 out of 57 composite faecal samples from small commercial farms and in 12 out of 43 composite faecal samples from small backyard holdings. The PCR products of 18 positive samples were sequenced for genotyping. PBoV sequences grouped into the PBoV-a, PBoV-b and PBoV-c groups with 90.81% to 99.25% nucleotide identity. All Croatian PBoV sequences showed a high nucleotide and amino acid identity with PBoV sequences from China and Hong Kong, the United States, Sweden, and Slovenia. These results clearly show that PBoV is circulating among the domestic pig population in Croatia.
detect parasitic nucleic acids by polymerase chain reaction (PCR) including conventional PCR, nested PCR and quantitative PCR; and finally serological methods based on the detection of a specific IgG response against L. infantum , including enzyme
Specific oligonucleotide primers were selected and combined in a multiplex arrangement, in order to detect simultaneously three economically important porcine viruses by polymerase chain reaction (PCR). The pathogen panel was comprised of viruses that cause reproductive failure in infected herds: Aujeszky’s disease virus (ADV), porcine parvovirus (PPV) and porcine respiratory and reproductive syndrome virus (PRRSV). In order to reduce the time required for the detection of the pathogens, the assay was optimised to a RapidCycler PCR instrument. The multiplex PCR assay was shown to be specific, sensitive and rapid, because the results were read in less than 60 min after sample preparation. Due to its speed, efficiency and sensitivity, the described rapid multiplex PCR assay serves as a useful novel tool in the veterinary diagnostic laboratories for the quick and complex detection of these important porcine pathogens.
For the molecular detection of Treponema pallidum authors introduced and used a nested PCR amplifying a conservative portion of the gene coding for the Tp 47 kDa membrane protein. PCR verified the presence of T. pallidum specific DNA in 5.7 per cent of syphilis seronegative 105 MSM belonging to HIV risk group. Treponema DNA was also detected in HIV infected, syphilis seronegative cases. Specificity of the method was demonstrated in rabbit inoculation test and also in clinically positive syphilis cases.
Encephalitozoon intestinalis infection of sows is reported from a pig farm in Slovakia. Spores were detected by direct microscopic visualisation in the faeces of 25 out of 27 sows (92.6%). This finding was also supported serologically by the presence of specific anti-E. intestinalis antibodies and by a species-specific polymerase chain reaction (PCR). This is the first report on E. intestinalis infection of swine in Europe.
Small ruminant lentiviruses (SRLV) are spread throughout the world, including Slovenia, where the first evidence of caprine arthritis encephalitis virus (CAEV) infection was found in 1996. This study was conducted to investigate the molecular and genetic characteristics of SRLV infection in Slovenia in order to classify our strains in relation to other known SRLV strains worldwide as well as to establish molecular techniques in concordance with serology. In this study, 340 goats and sheep were tested. Serological examination revealed that 57% of the goats and only 14% of the sheep were seropositive. The results of this study also show that the polymerase chain reaction (PCR) used in this study is less reliable than ELISA, with only 60.6% of the seropositive animals being PCR positive. Thirty-eight nucleotide sequences of the gag region encoding the matrix protein were determined and compared to sequences derived from the GenBank, revealing that Slovenian SRLV strains belong to sequence groups A and B, being maedivisna virus (MVV) and CAEV-like, respectively. In one goat herd, the presence of more than one genotype was confirmed and the majority of goat SRLV sequences were more closely related to MVV than to CAEV prototype strains.
Equine piroplasmosis in donkeys has been recognised as a serious problem of major economic importance. The present molecular study is the first investigation of the presence of Theileria equi and Babesia caballi in Balkan donkeys and of the possible haematological alterations related to it. A total of 70 apparently healthy donkeys from Serbia were included in this study. The overall prevalence of T. equi infection in donkeys tested with multiplex PCR was 50%. There was no B. caballi-positive sample. Infections in donkeys included in this study seem to be associated with decreased red blood cell count, haemoglobin concentration, haematocrit and platelet count, and with increased white blood cell count, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Altered haematological parameters in donkeys can lead to a decrease in working capacity and production performance. Further molecular research and long-term monitoring of equine piroplasmosis is needed in Serbia and throughout Europe.
on real-time RT-PCR (RT-qPCR), are the main tools for the detection of SARS-CoV-2, the diagnosis was initially centralized at reference hospitals, with well-equipped microbiology departments and qualified staff [ 2 , 3 ]. Within a few weeks, new
