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Abstract

Small ruminant lentivirus (SRLV), which causes caprine arthritis encephalitis in goats and ovine progressive pneumonia (maedi-visna disease) in sheep, is classified in genus Lentiviruses belonging to Retroviridae family. It persists in infected goats and sheep, which mostly are sub- clinical. A serological survey was conducted to determine the prevalence of small ruminant lentivirus infection in Thai goat population. Serum samples were taken from 1,925 goats distributed throughout the country, then they were tested for the presence of SRLV antibodies using commer- cial indirect enzyme-linked immunosorbent assay (ELISA) test kits. Results revealed that a total of 68 goats were found seropositive, representing the apparent prevalence and true prevalence of 3.57% and 2.60%, respectively. The seroprevalence, revealed in this study, was lower than in the previous reports. The decreasing of seroprevalence might be caused by successful control strategies from Department of Livestock Development (DLD).
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Authors and Affiliations

T. Mongkonwattanaporn
1
P. Lertwatcharasarakul
2
P. Intaravichai
3
T. Rukkwamsuk
1

  1. Department of Large Animal and Wildlife Clinical Sciences, Faculty of Veterinary Medicine, Kasetsart University, 1 Moo 6, Malaiman Road, Kamphaeng Saen, Nakhon Pathom 73140, Thailand
  2. Department of Pathology, Faculty of Veterinary Medicine, Kasetsart University, 1 Moo 6, Malaiman Road, Kamphaeng Saen, Nakhon Pathom 73140, Thailand
  3. Faculty of Veterinary Medicine and Applied Zoology, HRH Princess Chulabhorn Collage of Medical Science, Chulabhorn Royal Academy, 906 Kamphaeng Phet 6 Road, Talat Bang Khen, Lak Si, Bangkok 10210, Thailand
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Abstract

Porcine parvovirus (PPV) is a major causative agent in reproductive pig disease. The swine industry faces a significant economic and epizootic threat; thus, finding a reliable, quick, and practical way to detect it is essential. In this investigation, recombinant PPV VP2 protein was expressed in the Escherichia coli ( E. coli) expression systems. As shown by electron microscopy (TEM), Western blot, and hemagglutination (HA) assays, the recombinant VP2 protein was successfully assembled into virus-like particles (VLPs) after being expressed and purified. These VLPs had a structure that was similar to that of real PPV viruses and also exhibited HA activity. These VLPs induced high levels of PPV-specific antibody titers in mice after immunization, indicating that the VLPs may be beneficial as potential candidate antigens. VLPs were used as the coating antigens for the VLP ELISA, and the PPV VLPs-based ELISA displayed a high sensitivity (99%), specificity (93.0%) and agreement rate (98.3%) compared to HI assay, and the agreement rate of this ELISA was 97.5% compared to a commercial ELISA kit. Within a plate, the coefficient of variation (CV) was 10%, and between ELISA plates, the CV was 15%. According to a cross-reactivity assay, the technique was PPV-specific in contrast to other viral illness sera. The PPV VLP indirect-ELISA test for PPV detection in pigs with an inactivated vaccine showed that the PPV-positive rate varied among different sample sources from 88.2 to 89.6%. Our results indicate that this ELISA technique was quick, accurate, and repeatable and may be used for extensive serological research on PPV antibodies in pigs.
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Bibliography

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Authors and Affiliations

Y. Li
1
Q. Wang
2
W. Yue
1
X. Li
1
Y. Chen
1
Y. Gao
1

  1. Beijing Biomedicine Technology Center of JoFunHwa Biotechnology (Nanjing Co. Ltd.); No.25 Xiangrui Street Daxing District, Beijing 102600 China
  2. State Key Laboratory for Animal Disease Control and Prevention, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Harbin 150001, China
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Abstract

Progesterone (P4) is responsible for the main reproduction processes. Concentration of P4 varies widely among different determination methods, and interpretation of these values may be difficult. The objective of the current study was to assess the agreement of three different enzyme immunoassays (ELISA) in relation to radioimmunoassay (RIA) of P4 concentration assessment of beef cow serum samples. Samples were collected randomly considering high (pregnant cows) and low (non-pregnant cows) P4 concentrations. Depending on the P4 assessment method, four groups were created as follows: Group 1 – direct samples assessed by ELISA, Group 2 – extracted samples assessed by ELISA, Group 3 – samples assessed by automated ELISA, and Group 4 – samples assessed by RIA.

The mean progesterone concentration was 4.50 ng/mL, 1.24 ng/mL, 4.07 ng/mL and 4.39 ng/mL from Group 1 to Group 4, respectively. The mean difference (MD) between Group 1, Group 2 and Group 3 individually compared with Group 4 was −0.10 ± 1.24 ng/mL, 3.15 ± 3.58 ng/mL and 0.33 ± 1.42 ng/mL, and the 95% confidence interval (CI) for the differences (s) was from −0.99 to 0.78 ng/mL, from 0.59 to 5.71 ng/mL, and from −0.69 to 1.34 ng/mL, respectively. The confidence interval for the lower and upper limit of the agreement ranged from −4.12 to −1.05 ng/mL and from 0.84 to 3.91 ng/mL between Group 1 and Group 4, from −8.45 to 0.42 ng/ mL and from 5.88 to 14.75 ng/mL between Group 2 and Group 4, from −4.29 to −0.76 ng/mL, and from 1.41 to 4.94 ng/mL between Group 3 and Group 4.

Our findings show that the best agreement with RIA was observed for Group 1 and Group 3, while the agreement in the extraction method was least accurate.

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Authors and Affiliations

A. Rekešiūtė
A. Januškauskas
A. Šiukščius
V. Riškevičienė
A. Vitkauskienė
R. Verkauskienė
R. Antanaitis
H. Žilinskas
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Abstract

There are several infectious agents of domestic cattle that can also be present in free-living ruminant populations. These include bovine herpesvirus 1 (BoHV-1) and bovine viral diarrhea virus (BVDV) which are the causative agents of infectious bovine rhinotracheitis and bovine viral diarrhea, respectively. The study was conducted on serum samples from 59 red deer, 24 roe deer, and 3 fallow deer (86 in total), originating from two geographically separate areas of Poland. The samples were tested with commercially available ELISA tests for BoHV-1 and BVDV. The overall seroprevalence was 5.8% and 3.5%, respectively. All positive samples originated exclusively from red deer. Because of BoHV-1 ELISA cross reactivity with cervid herpesvirus 1 and 2 (CvHV-1 and -2) the nature of alphaherpesviruses infecting the sampled animals could not be assessed.

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Authors and Affiliations

M. Fabisiak
A. Sałamaszyńska
T. Stadejek
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Abstract

Avian encephalomyelitis (AE) is a viral disease of poultry. Although the disease has a milder clinical course in turkeys than in chickens, reproductive flocks of turkeys are vaccinated against AE. Commercial AE ELISA kits are specifically designed for chickens, which makes it difficult to implement these tests in serological monitoring of turkey flocks. The aim of the study was to compare the AE serological results provided by two ELISA kits from different producers when testing an AE-vaccinated flock of turkey hens and their progeny. We detected differences in the sensitivity of the ELISAs for testing specific anti-AE antibody levels in turkey serum samples.
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Authors and Affiliations

M. Śmiałek
1
J. Kowalczyk
1
B. Ogonowska-Woźniak
2
A. Koncicki
1

  1. Department of Poultry Diseases, Faculty of Veterinary Medicine, University of Warmia and Mazury, Oczapowskiego 13, 10-719 Olsztyn, Poland
  2. Hybrid Turkeys, Budowlana 2A, 10-424 Olsztyn, Poland
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Abstract

The aims of this study were to evaluate the prevalence of Coxiella burnetii on both herd and animal level based on ELISA and PCR tests. Antibodies to C. burnetii were detected in 22 out of the 24 bulk tank milk samples (91.6%) tested by ELISA and the IS1111 element of C. burnetii was detected in 10 out of the 24 samples (41.6%) by real-time polymerase chain reaction (PCR). ELISA testing showed individual seropositivity in 67 out of the 165 cows (40.6%) examined in 24 dairy cattle farms in different parts of the Czech Republic. Our study revealed that the prevalence of C. burnetii has increased substantially in the Czech Republic over the past 30 years, and that the causative agent is a potential risk factor for some reproductive problems in dairy farms and a possible risk factor for human infection.
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Bibliography


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Authors and Affiliations

A. Dobos
1
I. Fodor
1
T. Tekin
2
D. Đuričić
3
M. Samardzija
4

  1. CEVA-Phylaxia Co. Ltd., Szállás u. 5, Budapest, H-1107, Hungary
  2. Ceva Animal Health Slovakia, Sro. Račianska 153, 831 53 Bratislava, Slovakia
  3. Mount-Trad d.o.o., Industrijska 13, 43280, Garešnica, Croatia
  4. Faculty of Veterinary Medicine University of Zagreb, Heinzelova 55, 10000 Zagreb, Croatia
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Abstract

Several pre-analytical factors may influence the accurate measurements of testosterone (T) and therefore, these factors must be a significant concern. This study aimed to examine the effects of 1) time of sample collection, 2) delay to centrifugation, 3) sample matrix types, and 4) device and duration of sample storage on the T concentrations. Blood samples were collected from 34 bucks of Kacang goats. For testing the effect of collection time, 12 pairs of morning and afternoon samples were collected. For testing the effect of delayed centrifugation, 24 samples were subjected to treatments: (i) centrifuged < 1 hour after collection (control group), (ii) centrifuged 6, 12, and 24 hours after collection (test groups). For testing the different sample matrix types, 10 samples were processed as serum and plasma. For testing the effect of sample storage device and duration, 60 samples were subjected to treatments: i) frozen at -20OC (control group), ii) stored in a cooler box, a styrofoam box, and a thermos-flask for two, four, and six days (test groups). T concentrations were measured using a validated testosterone ELISA kit. Concentrations of plasma testosterone (pT) from morning samples were significantly higher compared to afternoon samples (p<0.05). Delayed centrifugation for up to 24 h decreased significantly on pT concentrations (p<0.05). The concentrations of T from serum and plasma did not differ and showed a strong correlation (r=0.981). Storage device and duration affected the T concentrations compared to frozen samples (p<0.05) which T concentrations were stable for up to 4 days in a styrofoam box and a thermos-flask and up to 6 days in a cooler box. In conclusion, the measurement accuracy and stability of T concentrations in goats are affected by collection time, delay to centrifugation, and device and duration of storage.
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Authors and Affiliations

G. Gholib
1
S. Wahyuni
1
A. Abdilla
1
T.P. Nugraha

  1. Faculty of Veterinary Medicine, Universitas Syiah Kuala, Banda Aceh, 23111, Aceh, Indonesia
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Abstract

The equine infectious anaemia virus (EIAV) is one of the most serious equine diseases worldwide. There is scarce information on the epizootiology of equine infectious anaemia (EIA) in Saudi Arabia. Given the importance of the equine industry in Saudi Arabia, this cross- -sectional study aims to provide information about the prevalence of EIAV based on serological surveillance of the equine population in the country. A total of 4728 sera samples were collected (4523 horses and 205 donkeys) between December 2017 and November 2019. All samples were tested using commercially available EIAV ELISA. All tested samples showed negative results for EIAV antibodies with a 95% confidence interval. The results provided evidence that Saudi Arabia’s equine populations (horses and donkeys) are currently free of EIAV. The results also suggest the need for continuous monitoring of EIAV and strict regulation when importing horses from other countries.
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Bibliography


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Authors and Affiliations

S. Kasem
1 2
O. Hashim
1
A. Alkarar
1
A. Hodhod
1 3
A. Elias
1
M. Abdallah
1
A. Al-Sahaf
1
A. Al-Doweriej
1
I. Qasim
1
A.S. Abdel-Moneim
4

  1. Ministry of Environment, Water and Agriculture, 65 King Abdulaziz Road, Riyadh, 11195, Saudi Arabia
  2. Department of Virology, Faculty of Veterinary Medicine, Kafrelsheikh University, Kafrelsheikh, El Geish Street, 33516, Egypt
  3. Animal Health Research Institute – Virology Department – Damanhur Branch – Egypt
  4. Microbiology Department, Virology Division, College of Medicine, Taif University, Al-Taif, Saudi Arabia
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Abstract

The tuber necrotic strain of Potato virus Y (PVYNTN) causes widespread disease and has severe negative effects on the growth and yields of plants, especially those of the Solanaceae family. The consequences of residual toxicity and non-biodegradation of synthetic chemicals and pollution of the environment has led to investigations into new non-toxic and biological treatments to control plant viral diseases. Ethanolic extracts of Bowiea volubilis (bulbs), Cotyledon orbiculata (leaves), Gomphocarpus fruticosus (leaves), Merwilla plumbea (dry and fresh bulbs), Nerium oleander (leaves), and the fruits and leaves of Strophanthus speciosus, were evaluated against PVYNTN in vivo and in vitro. At a concentration of 20 mg · ml−1, ethanolic extracts of Strophanthus speciosus (leaves) and fruits (50 mg · ml−1) significantly reduced the expression of PVYNTN symptoms on tobacco plants in vitro without affecting the normal growth and development of the plant. Similarly, at 50 mg · ml−1, N. oleander, C. orbiculata and B. volubilis (fresh bulbs) and S. speciousus leaves at 20 mg · ml−1 extracts showed significant differences in PVYNTN symptoms in the in vivo experiment. Strophanthus speciosus leaf and fruit extracts showed significant inhibition in the in vitro and in vivo assays and demonstrated that S. speciosus has potential to be used as an antiphytoviral treatment.

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Authors and Affiliations

Simin Emamzadeh Yazdi
Julia Mulabisana
Gerhard Prinsloo
Michele Cloete
Quenton Kritzinger
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Abstract

The study was aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA), which can detect specifically Feline herpesvirus type 1 (FHV-1). The primers were designed based on the conserved sequence of FHV-1 glycoprotein B gene. The recombinant protein with reactogenicity was purified as coating antigen of the assay. The indirect ELISA, characterized by high sensitivity showed no cross-reaction with two types of feline virus, had detection limit at 1:2000 dilution. The positive rate of the assay, according to the determined cutoff value (0.25), was basically consistent with Feline Herpes Virus Antibody ELISA kit. In conclusion, the indirect ELISA with high repeatability and reproducibility can be used for detecting FHV-1, and can provide necessary support to related research.

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Authors and Affiliations

Y. Tan
G. Dong
J. Niu
Y. Guo
S. Yi
M. Sun
K. Wang
G. Hu
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Abstract

Field surveys were undertaken in 1997–1999 across five ecological zones in Nigeria to collect isolates of Maize streak virus (MSV), genus Mastrevirus. Apart from maize (Zea mays L.), 15 other grass species were found with MSV symptoms in Nigeria. These hosts showed two types of symptoms viz: mild (with or without mottle) or severe (typical symptoms in maize). When Cicadulina storeyi China was used to attempt transmission of these isolates of MSV to seedlings a susceptible maize hybrid CML 254 X CML 247, six isolates were not transmissible to maize. Seven isolates that were transmissible to maize produced mild symptoms. The viral agents causing typical or severe streak symptoms in Axonopus compressus (Sw.) P. Beauv., Brachiaria distichophylla (Trn.) Stapf, Dactyloctenium aegyptium (Linn.) P. Beauv. and Setaria barbata (Lam.) Kunth produced symptoms that were typical of MSV in farmers fields, when transmitted to maize. Out of 33 plant species that seedlings were challenged with MSV, only eight proved susceptible. Four of them showed mild symptoms while the other four showed severe symptoms of MSV. Only three isolates collected during the surveys did not react with a MSV polyclonal antiserum produced in mice in Double Antibody Sandwich-Enzyme-Linked Immunosorbent Assay (DAS-ELISA). These isolates were found in Andropogon gayanus Kunth (from Kaduna), Thelepogon elegans Roth ex Toem & Schult (from Kadawa) and Rottboellia cochinchinensis (Lour.) Clayton (from Jos) exhibited mild streak/mottle symptoms. Specific monoclonal antibodies, raised against MSV, reacted with 12 out of 25 samples tested. The DAS-ELISA data also showed significant variation in concentration of the virus in the different plant hosts. The relationship dendogram through SDS-PAGE among eight purified virus isolates show 55–90% variation. At 0.55 coefficient of similarity, the dendogram divided the samples into two groups while at 0.9 coefficient of similarity, the 8 isolates were identified as distinct genetic entities.

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Authors and Affiliations

Sunday Oluwafemi
G. Thottappilly
Matthew D. Alegbejo
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Abstract

Immunoaffinity chromatography (IAC) is a fundamental isolation and purification tool which is incorporated in a substantial range of therapeutic and diagnostic applications. This study has reappraised the usefulness of immunoaffinity chromatography for the purification of polyclonal antibodies. Protein A based IAC is a convenient and reliable method for purification of IgG, from hyperimmunesera (HIS) raised in experimental animals such as rabbits, guinea pigs and mice to be utilized in pharmaceutics and diagnostics. The 146S fraction of Foot and Mouth Disease virus (FMDV) TCID50=10 5.6 was cultured on a baby hamster kidney cell line 21 (BHK-21), concentrated using salt precipitation method using PEG 6000, purified by size exclusion chromatography (SEC) using Sepharose-30 at 254nm absorbance. Purification of 146S FMDV was analyzed using 12% SDS-PAGE which provided two bands of light and heavy chains. The alum-based vaccine, consisting of ≥10μg of 146S FMDV, was applied in 10 male rabbits and 10 male guinea pigs and two animals of each group were taken as a negative control. The titer of serum was calculated using virus neutralization test. A Protein-A kit (Thermo scientific- 44667, 0528.2) was used to purify HIS raised against 146S FMDV and validated using 12% SDS PAGE in reducing condition. The data demonstrate that protein-A affinity chromatography is an efficient tool for the purification of antibodies from hyper-immune sera raised against 146S FMDV and can be used for the production of diagnostic kits e.g. Enzyme linked immuno-sorbent assay (ELISA) and radioimmunoassay.
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Authors and Affiliations

A. Munir
1
A.A. Anjum
1
I. Altaf
2
A.R. Awan
3

  1. Institute of Microbiology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Outfall road, Lahore, Pakistan
  2. Quality Operations Laboratory, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Outfall road, Lahore, Pakistan
  3. Department of Biochemistry and Biotechnology, Faculty of Veterinary Sciences, University of Veterinary and Animal Sciences, Outfall road, Lahore, Pakistan
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Abstract

In the spring of 2019, many plants, mainly winter wheat, were observed to have dwarfism and leaf yellowing symptoms. These plants from several regions of Poland were collected and sent to the Plant Disease Clinic of the Institute of Plant Protection – National Research Institute in Poznań to test for the presence of viral diseases. Double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) results showed numerous cases of Wheat dwarf virus (WDV) and a few cases of plant infections caused by Barley yellow dwarf viruses (BYDVs). WDV was detected in 163 out of 236 tested winter wheat plants (69.1%), in 10 out of 27 tested winter barley plants (37%) and in 6 out of 7 triticale plants (85.7%) while BYDVs were found, respectively, in 9.7% (23 out of 236) and in 18.5% (5 out of 27) of tested winter forms of wheat and barley plants. Infected plants came mainly from the regions of Lower Silesia and Greater Poland. Furthermore, individual cases of infections were also confirmed in the following districts: Lubusz, Opole, Silesia, Kuyavia-Pomerania and Warmia-Masuria. Results of Duplex-immunocapture-polymerase chain reaction (Duplex-IC-PCR) indicated the dominance of WDV-W form in wheat and WDV-B form in barley plants. Moreover, results of reverse transcription – polymerase chain reaction (RT-PCR) connected with restriction fragment length polymorphism (RFLP) analysis, performed for 17 BYDVs samples, revealed 8 BYDV-PAS, 4 BYDV-MAV and 2 BYDVPAV as well as the presence of two mixed infections of BYDV-MAV/-PAS and one case of BYDV-MAV/-PAV. Next, RT-PCR reactions confirmed single BYDV-GAV infection and the common presence of BYDV-SGV. To the best of our knowledge, in 2020 the viruses were not a big threat to cereal crops in Poland.

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Authors and Affiliations

Katarzyna Trzmiel
ORCID: ORCID

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