This Open Access volume explains how major nuclear and radiological emergencies (NREs) can have implications at local, national and international level. The response to NREs requires a competent ...decision-making structure, clear communication and effective information exchange. National veterinary services have the responsibility to plan, design and manage animal production system in their countries. These activities cover animal health, animal movement control, production control and improvement, and control of the products of animal origin before their placement on the market. Release of radionuclides after NREs can cause substantial contamination in the animal production systems. Critical responsibility of veterinary authorities is therefore to prevent such contamination, establish early response mechanisms to mitigate the consequences and prevent placement of contaminated products of animal origin on the market for human consumption. This work summarizes the critical technical points for effective management of NREs for national veterinary services.
Although rabies is enzootic in the Democratic Republic of the Congo, there is very little molecular epidemiological information about the viruses circulating in animals. In this study, a fragment of ...the rabies virus (RABV) nucleoprotein gene was amplified and sequenced from 21 animal brain samples collected in two western provinces of the country between 2008 and 2017. The samples tested were from cat (
n
= 1), dog (
n
= 17), goat (
n
= 2), and sheep (
n
= 1). Phylogenetic analysis revealed that the sequences generated were highly similar to each other and belonged to lineage Africa 1b clustering with a single sample identified in a canine in the Republic of Congo in 2014. This is the first molecular epidemiological study of RABV in the DRC and the data generated will assist authorities in the development of effective control strategies for rabies in the country.
Poxviruses within the Capripoxvirus, Orthopoxvirus, and Parapoxvirus genera can infect livestock, with the two former having zoonotic importance. In addition, they induce similar clinical symptoms in ...common host species, creating a challenge for diagnosis. Although endemic in the country, poxvirus infections of small ruminants and cattle have received little attention in Botswana, with no prior use of molecular tools to diagnose and characterize the pathogens.
A high-resolution melting (HRM) assay was used to detect and differentiate poxviruses in skin biopsy and skin scab samples from four cattle, one sheep, and one goat. Molecular characterization of capripoxviruses and parapoxviruses was undertaken by sequence analysis of RPO30 and GPCR genes.
The HRM assay revealed lumpy skin disease virus (LSDV) in three cattle samples, pseudocowpox virus (PCPV) in one cattle sample, and orf virus (ORFV) in one goat and one sheep sample. The phylogenetic analyses, based on the RPO30 and GPCR multiple sequence alignments showed that the LSDV sequences of Botswana were similar to common LSDV field isolates encountered in Africa, Asia, and Europe. The Botswana PCPV presented unique features and clustered between camel and cattle PCPV isolates. The Botswana ORFV sequence isolated from goat differed from the ORFV sequence isolated from sheep.
This study is the first report on the genetic characterization of poxvirus diseases circulating in cattle, goats, and sheep in Botswana. It shows the importance of molecular methods to differentially diagnose poxvirus diseases of ruminants.
Celotno besedilo
Dostopno za:
DOBA, IZUM, KILJ, NUK, PILJ, PNG, SAZU, SIK, UILJ, UKNU, UL, UM, UPUK
Aim of this study was to characterize pigeon variant of Newcastle disease virus (NDV) isolated from backyard poultry using classical and molecular methods. In standard hemagglutination inhibition ...(HI) test both polyclonal NDV antiserum and monoclonal antibodies 161/617 specific for pigeon variants of NDV showed inhibition of heamagglutination of the isolated virus. Intracerebral pathogenicity index (ICPI) has shown that the isolate is mesogenic virus (ICPI = 0.81). One-step RT-qPCR for detection of M gene was performed indicating a presence of NDV and RT-qPCR for discrimination between lentogenic and velogenic strains based on F gene was also performed indicating a presence of virulent NDV. A portion of the F gene was amplified and sequenced for determination of virulence and phylogenetic characterization. The F protein cleavage site sequence of the isolate had multiple basic amino acids at residues 112–116 and a phenyl alanine at residue 117 (112RRQKR*F117) which is typical for velogenic strains. The nucleotide sequence of 374 bp was aligned to begin at nt 47 and finish at 420 immediately after the cleavage site and compared with other reference strains from the region and worldwide. In the phylogenetic tree, the isolate clustered into genotype VIb, typical for PPMV-1. This strain is phylogenetically very similar to other PPMV-1 isolated from pigeons in Macedonia. Poultry infected with PPMV-1 can spread the virus in the absence of clinical signs, thus PPMV-1’s are constant threat to domestic poultry. This is the first report of evidenced spillover of PPMV-1 into poultry in Macedonia..
Equine infectious anemia (EIA) is an important viral disease characterized by persistent infection in equids worldwide. Most EIA cases are life-long virus carriers with low antibody reactions and ...without the appearance of clinical symptoms. A serological test with high sensitivity and specificity is required to detect inapparent infection. In this study, a B-cell common epitope-based blocking ELISA (bELISA) was developed using a monoclonal antibody together with the EIAV p26 protein labelled with HRP. The test has been evaluated against the standard and with field serum samples globally. This bELISA test can be completed within 75 min, and the sensitivity is higher than those of either the AGID or one commercial cELISA kit. This bELISA assay was 8–16 times more analytically sensitive than AGID, and 2 to 4 times more analytically sensitive than one cELISA kit by testing three sera from the USA, Argentina, and China, respectively. The 353 serum samples from Argentina were tested, in comparison with AGID, the diagnostic sensitivity and specificity of our bELISA assay were 100% (154/154) and 97.0% (193/199), respectively, and the accuracy of the bELISA test was 98.3%. The bELISA test developed in this study is a rapid, sensitive, specific method for the detection of EIAV infection, and could be a promising candidate for use in the monitoring of the EIA epidemic worldwide.
Key points
• A universal epitope-based blocking enzyme-linked immunosorbent assay (bELISA) was developed for detection of antibodies to EIAV.
• The bELISA assay can be used to test EIAV serum samples from different regions of the world including North America, South America, Europe, and Asia.
• The bELISA assay was evaluated in three different international labs and showed a better performance than other commercial kits.
•Eleven molecular diagnostic assays compared in their ability to detect SARS-CoV-2 RNA.•Good to excellent agreement between assays confirmed.•Valid alternatives to ad hoc molecular diagnostic assays ...identified.
Transmission mitigation of SARS-CoV-2 requires the availability of accurate and sensitive detection methods. There are several commercial ad hoc molecular diagnostic kits currently on the market, many of which have been evaluated by different groups. However, in low resource settings the availability and cost of these commercial kits can be a limiting factor for many diagnostic laboratories. In such cases alternatives need to be identified. With this in mind, eight commercial reverse transcription quantitative real-time PCR (RT-qPCR) master mixes from Applied Biosystems (Thermo Fisher Scientific), Bio-Rad, Biotech Rabbit, Promega, Qiagen, QuantaBio, Invitrogen (Thermo Fisher Scientific) and Takara using the same commercial primer and probe mix LightMix® Modular SARS and Wuhan CoV E-gene mix (TIB MolBiol, Germany) were evaluated. Three ad hoc molecular diagnostic kits GeneFinder™ COVID-19 Plus RealAmp kit (Osang Healthcare); genesig® Real-Time PCR Coronavirus COVID-19 (Primerdesign); and ViroReal® Kit SARS-CoV-2 & SARS-CoV (Ingenetix) were also included in the study. The limit of detection was calculated for each assay using serial dilutions of a defined clinical sample. The performances of the assays were compared using a panel of 178 clinical samples and their analytical specificity assessed against a panel of human betacoronaviruses. Inter assay agreement was assessed using statistical tests (Bland-Altman, Fleiss-Kappa and Cohen’s Kappa) and was shown to be excellent to good in all cases. We conclude that all of the assays evaluated in this study can be used for the routine detection of SARS-CoV-2 and that the RT-qPCR master mixes are a valid alternative to ad hoc molecular diagnostic kits.
African swine fever (ASF) is a highly lethal and contagious viral haemorrhagic disease of domestic and wild pigs, caused by the ASF virus (ASFV). After entering China in 2018, the disease has ...continued to spread through Asia. In September 2019, a team from the Indonesian Research Center for Veterinary Science, Bogor, investigated outbreaks in backyard pigs in the Dairi and Humbang Hasundutan districts of North Sumatra province. In January 2020, three pigs purchased from a pig seller in Bogor District, West Java province were also tested. Real‐time PCR results confirmed ASFV DNA in sixteen out of twenty‐nine samples, with nine positive samples from North Sumatra and seven from West Java. Four partial or full‐length genes (i.e. p72, p54, pB602L and CD2v) and a 356‐bp fragment between the I73R and I329L genes were sequenced from representative samples. Phylogenetic analysis established that the ASFV in the samples from both North Sumatra and West Java were identical, indicating a common source of infection, and that they belonged to the p72 genotype II and serogroup 8. The sequences from the Indonesian ASFVs were also identical to other genotype II ASFV from domestic pigs in Vietnam, China and Russia.
The objective of this study was to assess virulence using molecular methods and to phylogenetically characterize Newcastle disease virus (NDV) detected in common starlings (Sturnus vulgaris) in ...Macedonia. Nucleotide sequencing of the cleavage site of the F gene revealed an amino acid pattern specific for virulent strains of NDV with phenylalanine at position 117 and multiple basic amino acids between positions 112 and 116, i.e. sup.112RRQKR x FIG.sup.119. The 374 bp region of the fusion (F) gene used for phylogenetic analyses revealed that the detected strain belongs to subgenotype VIId of the class II NDV, and it is similar to virulent viruses detected in back-yard poultry in Macedonia in 2005 and 2006, as well as to viruses from poultry and wild birds detected in Serbia and Bulgaria in 2006 and 2007. Epidemiological data suggest that the common starlings were probably infected as a result of an ongoing epizootic in domestic poultry and they did not have a role in the primary introduction and spread of the virus to domestic poultry. This study represents the first report of the detection and molecular and phylogenetic characterization of virulent NDV from wild birds in Macedonia. Key words: common starling, Newcastle disease virus, molecular characterization Cilj ovog istrazivanja bio je molekularnim metodama odrediti virulenciju i filogenetska obiljezja virusa newcastleske bolesti (NB) dokazanog u cvorka (Sturnus vulgaris) u Makedoniji. Pokazalo se da nukleotidni slijed na mjestu cijepanja gena F daje aminokiselinski sastav specifican za virulentne sojeve virusa NB s fenilalaninom na poziciji 117 i uzastopnim bazicnim aminokiselinama izmedu pozicija 112 i 116, tj. sup.112RRQKR x FIG.sup.119. Filogenetska analiza podrucja od 374 bp fuzijskog (F) gena pokazala je da izdvojeni soj pripada podtipu VIId skupine II virusa NB, a srodan je virulentnim sojevima dokazanima u domace peradi u Makedoniji 2005. i 2006. kao i s virusima izdvojenima iz peradi i divljih ptica u Srbiji i Bugarskoj 2006. i 2007. Epizootioloski podatci upucuju na zakljucak da su cvorci vjerojatno bili zarazeni kao posljedica pojave NB u domace peradi te nisu imali nikakvu ulogu na pojavu i sirenje virusa na domacu perad. Ovo je prvo izvjesce o dokazu, molekularnim i filogenetskim obiljezjima virulentnog virusa NB u divljih ptica u Makedoniji. Kljucne rijeci: cvorak, virus newcastleske bolesti, molekulamakarakterizacija
Application of fluorescence based molecular assays for improved detection and typing of Brucella strains in clinical samples Krstevski, Kiril , Veterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of Macedonia; Naletoski, Ivancho , Animal Production and Health Section, Joint FAO/IAEA Division,International Atomic Energy Agency, Vienna, Austria; Mitrov, Dine , Veterinary Institute, Faculty of Veterinary Medicine, Ss. Cyril and Methodius University in Skopje, Republic of Macedonia ...
Macedonian veterinary review,
10/2015, Letnik:
38, Številka:
2
Journal Article
Recenzirano
Odprti dostop
Bacteria from the genus Brucella are causative agents of brucellosis – a zoonotic disease which affects many wild and domestic animal species and humans. Taking into account the significant ...socio-economic and public health impact of brucellosis, its control is of great importance for endemic areas. The chosen control strategy could be successful only if adapted to the current epidemiological situation. This implies that a choice of appropriate diagnostic procedures for detection and typing of Brucella spp. strains are of essential importance. Significant advancement of molecular techniques and their advantages compared to classical methods, give strong arguments in promotion of these techniques as a powerful tool for comprehensive diagnostics of brucellosis. Considering this, the major tasks of the study were to select and implement molecular tests for detection and genotyping Brucella spp. and evaluate their performances using DNA from cultivated brucellae (islolates) and limited number of tissue samples from seropositive animals. The obtained results confirmed that implemented real time PCR for Brucella spp. detection, as well as MLVA-16 used for genotyping, have excellent analytical sensitivity (4.2 fg of Brucella DNA were successfully detected and genotyped). Furthermore, compared to bacteriological cultivation of Brucella spp., real time PCR and MLVA-16 protocols showed superior diagnostic sensitivity and detected Brucella DNA in tissues from which Brucella could not be cultivated. Based on the summarized study results, we propose a diagnostic algorithm for detection and genotyping of Brucella spp. bacteria. Routine use of proposed diagnostic algorithm will improve the effectiveness of infection confirmation and help for accurate evaluation of epidemiological situation.
Cilj ovog istraživanja bio je molekularnim metodama odrediti virulenciju i filogenetska obilježja virusa newcastleske bolesti (NB) dokazanog u čvorka (Sturnus vulgaris) u Makedoniji. Pokazalo se da ...nukleotidni slijed na mjestu cijepanja gena F daje aminokiselinski sastav specifičan za virulentne sojeve virusa NB s fenilalaninom na poziciji 117 i uzastopnim bazičnim aminokiselinama između pozicija 112 i 116, tj. 112RRQKR*FIG119. Filogenetska analiza područja od 374 bp fuzijskog (F) gena pokazala je da izdvojeni soj pripada podtipu VIId skupine II virusa NB, a srodan je virulentnim sojevima dokazanima u domaće peradi u Makedoniji 2005. i 2006. kao i s virusima izdvojenima iz peradi i divljih ptica u Srbiji i Bugarskoj 2006. i 2007. Epizootiološki podatci upućuju na zaključak da su čvorci vjerojatno bili zaraženi kao posljedica pojave NB u domaće peradi te nisu imali nikakvu ulogu na pojavu i širenje virusa na domaću perad. Ovo je prvo izvješće o dokazu, molekularnim i filogenetskim obilježjima virulentnog virusa NB u divljih ptica u Makedoniji.
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