While mitogenome mutations leading to pathological manifestations are rare, more than 200 such mutations have been described in humans. In contrast, pathogenic mitogenome mutations are rare in ...domestic animals and have not been described at all in cattle. In the small local Slovenian cattle breed Cika, we identified (next-generation sequencing) two cows with the T10432C mitogenome mutation in the ND4L gene, which corresponds to the human T10663C mutation known to cause Leber's hereditary optic neuropathy (LHON). Pedigree analysis revealed that the cows in which the mutation was identified belong to two different maternal lineages with 217 individual cows born between 1997 and 2020. The identified mutation and its maternal inheritance were confirmed by Sanger sequencing across multiple generations, whereas no single analysis revealed evidence of heteroplasmy. A closer clinical examination of one cow with the T10432C mutation revealed exophthalmos, whereas histopathological examination revealed retinal ablations, subretinal oedema, and haemorrhage. The results of these analyses confirm the presence of mitochondrial mutation T10432C with homoplasmic maternal inheritance as well as clinical and histopathological signs similar to LHON in humans. Live animals with the mutation could be used as a suitable animal model that can improve our understanding of the pathogenesis of LHON and other mitochondriopathies.
Porcine epidemic diarrhoea (PED) is an emergent/re-emergent viral pig disease (caused by the virus belonging to the Coronaviridae family, in specific the Alphacoronavirus genus) of global importance. ...Clinical presentation is characterized with acute diarrhoea, vomiting and dehydration in pigs of all ages, with a possible high mortality in suckling piglets. The disease emerged in the USA in 2013 causing heavy losses, and re-emerged in Europe in 2014, but with milder consequences.
In the spring 2016, PED-like symptoms were reported to be seen on an agricultural holding in Eastern Croatia; laboratory workup confirmed the Croatia's first PED outbreak ever. Porcine epidemic diarrhoea virus (PEDV) strain responsible for the outbreak was of the S-INDEL genotype, much the same as other European PEDV strains. In 2017, a post-outbreak serology was carried out in three counties in Eastern Croatia, revealing seropositivity in pigs bred on four large industrial holdings (9.09%). The seroprevalence across PEDV-positive holdings was up to 82.8%. The latter holdings were unanimously managed by an enterprise that had never reported PED before.
PED has emerged in Croatian pig population causing potentially considerable losses. The circulating strain was of the S-INDEL genotype. Serological workup proved PEDV spread to another four agricultural holdings, demonstrating the importance of not only external, but also internal biosecurity measures.
To investigate the degree of genetic variability of bovine leukemia virus (BLV) strains circulating in Croatia, 29 isolates from the six largest dairy farms were examined by PCR for a segment of the ...gp51
env
gene, followed by DNA sequencing and phylogenetic analysis. The nucleotide sequences were compared with other previously characterized BLV strains from different geographical areas, comprising all seven known BLV genotypes. The Croatian sequences showed six to eight nucleotide substitutions: six silent substitutions and two amino acid changes. Four of those substitutions were within epitopes. In comparison to the sequences of other BLV genotypes, our isolates showed the closest relationship to genotype 1 isolates PL-3252 (FJ808585) and AL-148 (FJ808573) from Argentina. The degree of variation between our sequences and those of genotype 1 was 0.2- 4.6 %. In phylogenetic trees based on 400-nt and 519-nt sequences, all of the Croatian sequences clustered separately from the other sequences, revealing a new genotype.
In a total of 1536 blood serum samples analysed by ELISA, antibodies for IAV nucleoprotein (NP) were detected in 30.3%. Results from HI show that the most common subtype of swIAV in the Croatian pig ...population was H1N1 (44.6%), followed by H3N2 (42.7%) and H1N2 (26.3%). Antibodies to at least one subtype were detected in 62.19% of blood serum samples. Detection of swIAV antigen was performed by IHC and detected in 8 of 28 lung samples collected post-mortem. The matrix (M) gene was detected in nine of one hundred and forty-two lung tissue samples and in seven of twenty-nine nasopharyngeal swabs. Phylogenetic analysis of amplified HA and NA gene fragments in Croatian isolates suggests the presence of swIAV H1avN1av.
Porcine parvovirus 2 (PPV2) was detected in swine serum without showing any relationship with disease. The emergence of the virus seemed to be a unique event until other genetically highly similar ...parvoviruses were identified in China and, later in 2012, the presence of the virus was also described in Europe. PPV2 is widely distributed in pig populations where it is suspected to be involved in respiratory conditions, based on its frequent detection in lung samples. In order to investigate the potential pathogenic involvement of PPV2, 60 dead pigs were examined from two farms. They were necropsied and tested for PPV2 and PCV2 (Porcine circovirus type 2) by PCR; by Brown and Brenn (B&B) staining for bacteria; by immunohistochemistry (IHC) to detect CD3, Swine leukocyte antigen class II DQ (SLAIIDQ), lysozyme, porcine reproductive and respiratory syndrome virus (PRRSV), swine influenza (SIV), Mycoplasma hyopneumoniae (Mhyo); and by in situ hybridization (ISH) to detect ssDNA and dsDNA of PCV2. PPV2 positive samples were subjected to in situ polymerase chain reaction (IS-PCR) including double staining method to detect PPV2 and host cell markers. To calculate statistical difference we used GENMOD or LOGISTIC procedures in Statistical Analysis System (SAS®).
We found that the PPV2 was localized mostly in lymphocytes in lungs, lymph nodes and liver. Neither CD3 antigen nor lysozyme was expressed by these infected cells. In contrast, low levels of SLAIIDQ were expressed by infected cells, suggesting that PPV2 may have a specific tropism for immature B lymphocytes and/or NK lymphocytes though possibly not T lymphocytes.
The overall conclusion of this study indicates that PPV2 may contribute to the pathogenesis of pneumonia.
A total of 131 blood samples and 175 spleen samples were collected from three cervid species: roe deer (
), red deer (
) and fallow deer (
) inhabiting the continental part of Croatia. Serum samples ...were tested for antibodies against bovine herpesvirus 1, parainfluenza-3 virus, bluetongue virus, bovine respiratory syncytial virus, hepatitis E virus, bovine viral diarrhoea virus and enzootic bovine leukosis virus. The tested sera were negative for bovine viral diarrhoea virus, enzootic bovine leukosis virus, bluetongue virus, bovine respiratory syncytial virus and hepatitis E virus antybodies. The antibody prevalence in roe deer and red deer samples was 21.11% for bovine herpesvirus 1 and 75.55% for parainfluenza-3 virus. Sera from bovine herpesvirus 1 positive animals were subsequenty tested with comparative virus neutralization test and bovine herpesvirus 1 neutralising antibodies were found in 18 (out of 19) sera. In fallow deer, no antibodies against any of the viral pathogens were detected. All spleen samples tested for bovine viral diarrhoea virus and enzootic bovine leukosis virus came back negative, except for one red deer spleen sample found to be weakly diarrhoea virus-positive. Our findings provide the first information on the exposure of Croatia-inhabiting cervid species to viral pathogens, and could serve as valuable baseline data for future investigations regarding deer exposure to various pathogens and the distribution of diseases shared between wildlife and livestock. As of now, the epidemiology of these viruses in the Croatian cervid population has been only poorly understood, so that further research is recommended.
► Fluorescent antibody virus neutralisation test was modified. ► Muscle extract and thoracic liquid as samples in the follow-up of fox oral vaccination campaigns against rabies were evaluated. ► ...Cytotoxic effect on cells is completely prevented.
The virus neutralisation test is used for the quantitation of specific antibodies in serum samples. However, the success of the test depends on the quality of samples. In the case of poor quality samples, a cytotoxic effect can be observed and the results of the test can be compromised. Additionally, the cytotoxic effect limits the use of different substances, such as muscle extract or liquid from thoracic cavity (thoracic liquid), as a sample for the detection of rabies virus neutralising antibodies in the follow-up of fox oral vaccination campaigns. To eliminate the cytotoxic effect, a modified fluorescent antibody virus neutralisation (mFAVN) test was developed and evaluated. In the mFAVN test, inocula were removed after a 1h and the cytotoxic effect was prevented. According to the results obtained, the specificity of the mFAVN test compared to the FAVN test was 88.8% and the sensitivity was 94.4%. The diagnostic validity of the test was 0.99 (CI=0.98–1.00). To evaluate the possibility of using muscle extract and thoracic liquid as samples for the virus neutralisation test, 102 sera, muscle extract and thoracic liquid samples of dog origin were tested with the mFAVN test. The correlation between sera and muscle extracts was 87.9% (r=0.88, p<0.001). The correlation between sera and thoracic liquid was 94.2% (r=0.94, p<0.001). These findings indicated that both muscle extract and thoracic liquid could be used as samples for detection of rabies virus neutralising antibodies in the follow-up of oral vaccination campaigns. To evaluate the level of elimination of the cytotoxic effect, the 102 samples of sera, muscle extracts and thoracic liquid of dog origin were also tested in parallel using the mFAVN and FAVN tests. In the mFAVN test, no instance of cytotoxic effect was observed in the cells. In the FAVN test, two sera (1.9%), 35 muscle extracts (34.3%) and 56 thoracic liquid samples (54.9%) showed cytotoxic effect. The results of this study strongly suggest that cytotoxic effect can be eliminated completely from the rabies virus neutralising antibody detection tests used in the follow-up of oral vaccination campaigns and that very poor quality samples, such as muscle extract and thoracic liquid, can be used.
Ptičja influenca u divljih kanida Jungić, Andreja; Jemeršić, Lorena; Savić, Vladimir ...
Veterinarska stanica,
04/2024, Letnik:
55, Številka:
6
Journal Article, Paper
Recenzirano
Odprti dostop
The risk of cross-species transmission of infectious diseases and zoonoses has increased due to the adaptive evolution of pathogens and anthropogenic landscape changes. A current example is the avian ...influenza (AI) virus, which can infect not only avian species but also mammals. In these species, infections are often associated with severe neurological symptoms. AI viruses are primarily not well adapted to mammalian hosts and are not efficiently transmitted among them. To accomplish this, AI viruses must acquire adaptations or mutations that contribute to replication efficiency and virulence in mammals. The exact combination and interaction of mutations that result in optimal adaptation to mammals is still unknown and may vary between host species and virus subtypes. The globally increasing number of infected birds with highly pathogenic AI (HPAI) increases the likelihood of contact between birds and mammals, especially wildlife. One reason for the adaptation of HPAI to mammals is theexposure of wild mammals to infected birds or their carcasses. According to the current data, wild canids, such as red foxes, are among the most infected mammals. However, infections caused by neurotropic viruses, as well as HPAI, have been scarcely studied or not at all in jackals. Since jackals are scavengers that have spread rapidly in Europe and Croatia in recent years, surveillance of AI virus infection in these canids is of the utmost importance. Furthermore, interactions between wild carnivores and domestic animals are of concern, as infections with HPAI pose a public health threat due to the increased risk of mammalian adaptation.
Tijekom godina, rizik od međuvrsnog prijenosa zaraznih bolesti i zoonoza se zbog zbog adaptivne evolucije patogena i antropogenih promjena u okolišu povećao. Trenutačni primjer je virus influence ptica (IP), a koji osim ptica može zaraziti i sisavce u kojih su infekcije često povezane s teškim neurološkim simptomima. Virusi IP ponajprije nisu dobro prilagođeni sisavcima kao domaćinima i ne prenose se među njima učinkovito. Da bi to postigli, moraju se prilagoditi ili mutirati što pridonosi učinkovitijoj replikaciji i povećanju virulencije u sisavaca. Točna kombinacija i međudjelovanje mutacija koje rezultiraju optimalnom prilagodbom virusa IP na sisavce još uvijek je nepoznata i može varirati između domaćina i podtipova virusa. Vjerojatnost kontakta između ptica i sisavaca (posebice divljih) povećava se sa sve većim brojem zaraženih ptica visokopatogenim virusom IP (VPVIP) diljem svijeta. Jedan je od razloga prilagodbe VPVIP na sisavce i izlaganje divljih sisavaca zaraženim pticama ili njihovim lešinama. Među najugroženijim sisavcima su divlji kanidi (lisice i čagljevi). Neurotropni virusi, kao i VPVIP, su vrlo malo ili nikako istraženi u čagljeva. Uzevši u obzir povećanje broja i širenje staništa čagljeva u Europi, ali i u Hrvatskoj i činjenicu da se hrane lešinama, od iznimne je važnosti pratiti infekciju IP u ovih kanida. Infekcija VPVIP zbog povećanog rizika prilagodbe na sisavce predstavlja prijetnju javnom zdravlju što je posljedica sve češćeg kontakta ljudi, domaćih životinja i divljih zvijeri.
Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to ...successfully eradicate BLV through improved detection and management methods. However, with the increasing novel molecular detection methods there have been few efforts to standardize these results at global scale. This study aimed to determine the interlaboratory accuracy and agreement of 11 molecular tests in detecting BLV. Each qPCR/ddPCR method varied by target gene, primer design, DNA input and chemistries. DNA samples were extracted from blood of BLV-seropositive cattle and lyophilized to grant a better preservation during shipping to all participants around the globe. Twenty nine out of 44 samples were correctly identified by the 11 labs and all methods exhibited a diagnostic sensitivity between 74 and 100%. Agreement amongst different assays was linked to BLV copy numbers present in samples and the characteristics of each assay (i.e., BLV target sequence). Finally, the mean correlation value for all assays was within the range of strong correlation. This study highlights the importance of continuous need for standardization and harmonization amongst assays and the different participants. The results underscore the need of an international calibrator to estimate the efficiency (standard curve) of the different assays and improve quantitation accuracy. Additionally, this will inform future participants about the variability associated with emerging chemistries, methods, and technologies used to study BLV. Altogether, by improving tests performance worldwide it will positively aid in the eradication efforts.Bovine leukemia virus (BLV) is the etiological agent of enzootic bovine leukosis and causes a persistent infection that can leave cattle with no symptoms. Many countries have been able to successfully eradicate BLV through improved detection and management methods. However, with the increasing novel molecular detection methods there have been few efforts to standardize these results at global scale. This study aimed to determine the interlaboratory accuracy and agreement of 11 molecular tests in detecting BLV. Each qPCR/ddPCR method varied by target gene, primer design, DNA input and chemistries. DNA samples were extracted from blood of BLV-seropositive cattle and lyophilized to grant a better preservation during shipping to all participants around the globe. Twenty nine out of 44 samples were correctly identified by the 11 labs and all methods exhibited a diagnostic sensitivity between 74 and 100%. Agreement amongst different assays was linked to BLV copy numbers present in samples and the characteristics of each assay (i.e., BLV target sequence). Finally, the mean correlation value for all assays was within the range of strong correlation. This study highlights the importance of continuous need for standardization and harmonization amongst assays and the different participants. The results underscore the need of an international calibrator to estimate the efficiency (standard curve) of the different assays and improve quantitation accuracy. Additionally, this will inform future participants about the variability associated with emerging chemistries, methods, and technologies used to study BLV. Altogether, by improving tests performance worldwide it will positively aid in the eradication efforts.
Artritis-encefalitis koza (AEK) je neizlječiva virusna bolest proširena u većini zemalja koje se bave intenzivnim uzgojem koza. Ekonomsko značenje ove bolesti očituje se uginućem jaradi, smanjenim ...prirastom
i mliječnošću, prijevremenim lučenjem iz uzgoja. U razdoblju od 2012. do 2019. godine na području 16 županija u Republici Hrvatskoj
pretraženo je 10 715 uzoraka krvi, a tijekom tog osmogodišnjeg razdoblja specifična protutijela za virus AEK bila su potvrđena u
4 353 uzoraka krvnog seruma pa zaključno možemo govoriti o seroprevalenciji od 40,62 %. Rezultati genotipiziranja odabranih
sojeva LVMP ukazuju na cirkulaciju genetske skupine A i B u populaciji koza i to su prva takva istraživanja provedena u Hrvatskoj.
Dokaz prisutnosti genotipa A u stadu koza u kojem je istovremeno dokazan i genotip B ukazuje na činjenicu da sojevi LVMP nisu
ograničeni na jednu vrstu već da sojevi MVV koji se uglavnom nalaze u ovaca, cirkuliraju i u koza. Rezultati višegodišnjeg istraživanja
provedenog na Hrvaskom veterinarskom institutu ukazuju na široku rasprostranjenost virusa AEK u uzgojima koza u Republici Hrvatskoj. Ako znamo da jednom zaražena jedinka ostaje doživotni nositelj i prenositelj virusa postavlja se pitanje uspješnosti cijele proizvodnje pa je stoga jedini dugoročno održiv način uzgoja uzgoj slobodan od AEK.
PDF
