Nadzor influence u dvorišne peradi i ptica selica u Sloveniji

Blood samples and clocal swabs were taken from free-living migratory birds and backyard flock birds. Enzyme linked immunosorbent assays (ELISA) and haemagglutination inhibition test (HI) was used to screen free-living migratory birds and backyard flocks for antibodies against avian influenza viruses (AIV). Pools of cloacal swabs were used for the attempt of isolation of AIV. Samples were inoculated of 9 to 11 day-old chicken embryos. Amnioallantoic fluid were collected and analysed for haemagglutination activity. Pools of cloacal swabs were used for the detection of AIV by RT-PCR. Sera samples of four out of twenty three geese were positive on presence of specific antibodies against AIV. All attempts of virus isolation were negative. No AIV nucleic acid was detected by RT-PCR.Uzeti su uzorci krvi i obrisci nečisnica slobodnoživućih ptica selica i dvorišne peradi. Imunoenzimni test i inhibicija hemaglutinacije korišteni su kako bi se u slobodnoživućih ptica selica i dvorišne peradi utvrdila nazočnost protutijela za viruse influence ptica. Skupni uzorci briseva nečisnica korišteni su za izdvajanje virusa influence ptica. Uzorci su inokulirani u 9 do 11 dana stare kokošje embrije. Sakupljena je amnioalantoisna tekućina i pretražena na sposobnost hem-aglutinacije. Skupni uzorci brisova nečisnica korišteni su za dokaz virusa influence ptica pomoću RT-PCR. Četiri od 23 seruma gusaka bila su pozitivna na nazočnost specifičnih protutijela za virus influence ptica. Svi pokušaji izdvajanja virusa dali su negativan rezultat. Postupkom RT-PCR nije dokazana nazočnost nukleinske kiseline virusa influence ptica

Molekularni pristup identifikaciji virusa influence ptica

The recently raised awareness of the threat of a new influenza pandemic has stimulated the interest in the detection of influenza A viruses in the secretions of a wide variety of birds and mammals. Influenza A viruses are subtyped conventionally according to the characteristics of the external glycoproteins, haemagglutinin (HA) and neuraminidase (NA). All 16 HA and 9 NA subtypes have been isolated from aquatic birds that are believed to be a natural host and reservoir for influenza viruses. Poultry, specifically chickens and turkeys, are not considered to be a normal host for the virus, although transmission from wild birds to poultry occurs routinely. Although most HA subtypes have been found in poultry, particular emphasis is placed on the H5 and H7 HA subtypes of avian influenza virus because only these subtypes are known to cause highly pathogenic avian influenza (HPAI) in poultry. Recently developed molecular methods enable fast, accurate and reliable approach to identify influenza virus and its subtypes.Svijest o prijetnji nove pandemije influence potaknula je zanimanje za dokaz virusa influence A u sekretima različitih vrsta ptica i sisavaca. Virusi influence A uobičajeno se tipiziraju na osnovi osobina njihovih površinskih glikoproteina, hemaglutinina (HA) i neuraminidaze (NA). Svih 16 HA i devet NA podtipova izdvojeno je iz vodenih ptica koje su prirodni domaćin i rezervoar virusa influence. Perad, posebice kokoši i pure, ne smatra se uobičajenim domaćinom virusa iako se virus redovito s divljih ptica prenosi na perad. Premda su mnogi HA podtipovi izdvojeni iz peradi, posebna pozornost pridaje se podtipovima H5 i H7 virusa influence ptica, jer samo oni pripadaju visoko patogenim sojevima za perad. Nedavno razvijene molekularne metode omogućuju brzu, točnu i pouzdanu identifikaciju virusa influence i njegovih podtipova

Field efficacy of different vaccines against infectious bursal disease in broiler flocks

A field study was performed to determine the efficacy of three commercially available vaccines against infectious bursal disease (IBD) in commercial broilers raised in a high IBD virus (IBDV) risk area. Live attenuated intermediate and intermediate plus vaccines were used in four flocks. Birds were vaccinated orally at the estimated vaccination time. Three broiler flocks were vaccinated subcutaneously with a turkey herpesvirus (HVT)-IBD vector vaccine at one day old. Evaluation of the efficacy of different vaccines was focused on humoral immune response, bursa/body weight (B/Bw) ratio, molecular detection of IBDV in ileocaecal tonsils and bursa of Fabricius, and production parameters. The serological results showed that although the uptake of all three vaccine strains was confirmed in the lymphoid organs, no significant antibody response to vaccination was detected in flocks vaccinated with intermediate and intermediate plus vaccines. A significant increase in antibody titres detected in flocks vaccinated with the vector vaccine indicated its ability to induce an immune response in birds with a high level of maternally derived antibodies. Observations obtained in this field trial did not confirm the expected reduction of the B/Bw ratio in flocks vaccinated with less attenuated vaccines. No significant differences were observed between birds vaccinated with the vector vaccine and those immunised with the intermediate plus vaccine. Very virulent IBDV was confirmed in the flock vaccinated with the intermediate vaccine. The infection induced reduced B/Bw and moderate mortality but did not affect the production parameters. Field infection was not detected in broilers vaccinated with the intermediate plus vaccine and the vector vaccine

Detection of laryngotracheitis virus in poultry flocks with respiratory disorders in Slovenia

Abstract Infectious laryngotracheitis (ILT) is an acute, highly contagious infectious disease of the upper respiratory tract in chickens and other poultry species that causes significant economic losses in countries worldwide. Between 2017 and 2019, seven outbreaks of mild to severe respiratory disorders with high suspicion of ILT occurred in commercial and backyard poultry flocks in Slovenia. In all submissions, infection with ILT virus (ILTV) was confirmed by PCR, which is the first report of ILT in Slovenia. Circulating ILT strains were characterized by the sequence and phylogenetic analysis of two fragments of the ICP4 gene. Four strains—three detected in non-vaccinated flocks and one in a flock vaccinated against ILT—were identical or very similar to the chicken embryo–origin live virus vaccines, and the other three were closely related to Russian, Chinese, Australian, and American field strains and to tissue culture origin vaccine strains. As in other diseases, coinfections with other respiratory pathogens in confirmed ILT cases may cause a more severe condition and prolong the course of the disease. In our study, coinfections with Mycoplasma synoviae (7/7 tested flocks), infectious bronchitis virus (5/5 tested flocks), Mycoplasma gallisepticum (4/7 tested flocks), Ornithobacterium rhinotracheale (3/4 tested flocks), and avian pox virus (1/2 tested flocks) were confirmed, indicating the importance of these pathogens in the occurrence of ILT infections

Detection of herpesviruses in passerine birds captured during autumn migration in Slovenia

ABSTRACT: Herpesviruses (HVs) were detected by PCR in the cloacal swabs of 0.76% (4/525) clinically healthy free-living passerine birds from 32 different species captured in mist nets in Slovenia during the 2014 and 2017 autumn migrations. Herpesviruses were detected in the Eurasian Blackcap (Sylvia atricapilla), the Common Blackbird (Turdus merula), and the Eurasian Blue Tit (Cyanistes caeruleus). Phylogenetic analysis of partial DNA polymerase gene nucleotide sequences of the HV strains showed a distant relationship with other alphaherpesviruses of birds. In the phylogenetic tree, the HVs detected were clustered together with HV detected in Sulphur-crested Cockatoo and Neotropic Cormorants, as well as with known HVs such as gallid HV1, Psittacid HV1 and HV2, and passerine HV1. Different sequences of HVs with relatively low identity were detected in our study, suggesting that different HVs were circulating in passerines sampled during the autumn migration in Slovenia

Nadzor influence u dvorišne peradi i ptica selica u Sloveniji

Blood samples and clocal swabs were taken from free-living migratory birds and backyard flock birds. Enzyme linked immunosorbent assays (ELISA) and haemagglutination inhibition test (HI) was used to screen free-living migratory birds and backyard flocks for antibodies against avian influenza viruses (AIV). Pools of cloacal swabs were used for the attempt of isolation of AIV. Samples were inoculated of 9 to 11 day-old chicken embryos. Amnioallantoic fluid were collected and analysed for haemagglutination activity. Pools of cloacal swabs were used for the detection of AIV by RT-PCR. Sera samples of four out of twenty three geese were positive on presence of specific antibodies against AIV. All attempts of virus isolation were negative. No AIV nucleic acid was detected by RT-PCR.Uzeti su uzorci krvi i obrisci nečisnica slobodnoživućih ptica selica i dvorišne peradi. Imunoenzimni test i inhibicija hemaglutinacije korišteni su kako bi se u slobodnoživućih ptica selica i dvorišne peradi utvrdila nazočnost protutijela za viruse influence ptica. Skupni uzorci briseva nečisnica korišteni su za izdvajanje virusa influence ptica. Uzorci su inokulirani u 9 do 11 dana stare kokošje embrije. Sakupljena je amnioalantoisna tekućina i pretražena na sposobnost hem-aglutinacije. Skupni uzorci brisova nečisnica korišteni su za dokaz virusa influence ptica pomoću RT-PCR. Četiri od 23 seruma gusaka bila su pozitivna na nazočnost specifičnih protutijela za virus influence ptica. Svi pokušaji izdvajanja virusa dali su negativan rezultat. Postupkom RT-PCR nije dokazana nazočnost nukleinske kiseline virusa influence ptica

Molecular characterisation of infectious bursal disease viruses isolated in recent acute outbreaks in Solvenia

In 2004 and then in 2006 several outbreaks of infectious bursal disease (IBD) were reported in broiler and broiler breeder flocks in Slovenia. In this report ten recently emerged IBD viruses (IBDV) were characterised by sequence analysis of the VP2 hypervariable region and compared to previous Slovene IBDV strains from 1995/1996 and to some representative serotype 1 IBDV strains of different pathotypes. On the basis of nucleotide and amino acid identities, phylogenetic analyses and the presence of very virulent IBDV (vvIBDV) conserved amino acid substitutions, all Slovene isolates from recent outbreaks were identified as vvIBDV. Although some unique nucleotide exchanges and amino acid substitutions have been observed, the results of this study indicated that recent vvIBDV isolates are closely related with those from outbreaks in the 1990s. However, acute IBD has not been reported in commercial flocks in Slovenia for some years. This could lead to the conclusion that poor biosecurity and relaxed vaccination could be responsible for the re-emergence of vvIBDV

Rehoming and other refinements and replacement in procedures using golden hamsters in SARS-CoV-2 vaccine research

Effective vaccines are needed to fight the COVID-19 pandemic. Forty golden hamsters were inoculated with two promising vaccine candidates and eighteen animals were used in pilot trials with viral challenge. ELISA assays were performed to determine endpoint serum titres for specific antibodies and virus neutralisation tests were used to evaluate the efficacy of antibodies. All tests with serum from vaccinated hamsters were negative even after booster vaccinations and changes in vaccination protocol. We concluded that antibodies did not have sufficient neutralising properties. Refinements were observed at all steps, and the in vitro method (virus neutralisation test) presented a replacement measure and ultimately lead to a reduction in the total number of animals used in the project. The institutional animal welfare officer and institutional designated veterinarian approved the reuse or rehoming of the surplus animals. Simple socialization procedures were performed and ultimately 19 animals were rehomed, and feedback was collected. Recently, FELASA published recommendations for rehoming of animals used for scientific and educational purposes, with species-specific guidelines, including mice, rats, and rabbits. Based on our positive experience and feedback from adopters, we concluded that the rehoming of rodents, including hamsters, is not only possible, but highly recommended

Effects of coenzyme Q10 supplementation on oxidative stress markers, inflammatory markers, lymphocyte subpopulations, and clinical status in dogs with myxomatous mitral valve disease

Scarce data exist on the effects of coenzyme Q10 (CoQ10) supplementation in dogs with myxomatous mitral valve disease (MMVD). The purpose of this study was to investigate the effect of CoQ10 supplementation on oxidative stress markers (glutathione peroxidase, F2-isoprostanes), markers of inflammation (tumor necrosis factor-α, TNF soluble receptor II, leucocytes, and their subtypes), lymphocyte subpopulations (T helper and cytotoxic T lymphocytes, including activated T lymphocytes, and B lymphocytes), and echocardiographic and clinical parameters in dogs with MMVD. In this randomized, controlled, double-blind, longitudinal study, 43 MMVD dogs in stages ACVIM (American College of Veterinary Internal Medicine classification) B2 and ACVIM C and D (congestive heart failure (CHF)) received water-soluble coenzyme Q10 (100 mg twice daily) or placebo for 3 months, and 12 non-supplemented healthy dogs served as controls. All parameters were measured before and after supplementation in MMVD dogs and once in healthy dogs. CoQ10 supplementation had a positive impact on neutrophil percentage, lymphocyte percentage, and lymphocyte concentration in our cohort of dogs with CHF (ACVIM C and D). Conclusion: CoQ10 as an oral supplement may have benefits in terms of decreasing inflammation in dogs with MMVD and CHF

TXM peptides inhibit SARS-CoV-2 infection, syncytia formation, and lower inflamatory consequences

After three years of the SARS-CoV-2 pandemic, the search and availability of relatively low-cost benchtop therapeutics for people not at high risk for a severe disease are still ongoing. Although vaccines and new SARS-CoV-2 variants reduce the death toll, the long COVID-19 along with neurologic symptoms can develop and persist even after a mild initial infection. Reinfections, which further increase the risk of sequelae in multiple organ systems as well as the risk of death, continue to require caution. The spike protein of SARS-CoV-2 is an important target for both vaccines and therapeutics. The presence of disulfide bonds in the receptor binding domain (RBD) of the spike protein is essential for its binding to the human ACE2 receptor and cell entry. Here, we demonstrate that thiol-reducing peptides based on the active site of oxidoreductase thioredoxin 1, called thioredoxin mimetic (TXM) peptides, can prevent syncytia formation, SARS-CoV-2 entry into cells, and infection in a mouse model. We also show that TXM peptides inhibit the redox-sensitive HIV pseudotyped viral cell entry. These results support disulfide targeting as a common therapeutic strategy for treating infections caused by viruses using redox-sensitive fusion. Furthermore, TXM peptides exert anti-inflammatory properties by lowering the activation of NF-κB and IRF signaling pathways, mitogen-activated protein kinases (MAPKs) and lipopolysaccharide (LPS)-induced cytokines in mice. The antioxidant and anti-inflammatory effects of the TXM peptides, which also cross the blood-brain barrier, in combination with prevention of viral infections, may provide a beneficial clinical strategy to lower viral infections and mitigate severe consequences of COVID-19