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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


Agerholm JS (2013) Coxiella burnetii associated reproductive disorders in domestic animals - a critical review. Acta Vet Scand 55: 13.
Dobos A, Balla E (2021) Industrial dairy cattle farms in Hungary a source of Coxiella burnetii infection in humans. Vector Borne Zoonotic Dis. 21: 498-501.
Dobos A, Gábor G, Wehmann E, Dénes B, Póth-Szebenyi B, Kovács ÁB, Gyuranecz M (2020a) Serological screening for Coxiella burnetii in the context of early pregnancy loss in dairy cows. Acta Vet Hung 68: 305-309.
Dobos A, Kreizinger Z, Kovács ÁB, Gyuranecz M (2020b) Prevalence of Coxiella burnetii in Central and Eastern European dairy herds. Comp Immunol Microbiol Infect Dis 72: 101489.
Duron O, Sidi-Boumedine K, Rousset E, Moutailler S, Jourdain E (2015) The importance of ticks in Q fever transmission: what has (and has not) been demonstrated? Trends Parasitol. 31: 536-552.
ECDC (2019) European Centre for Disease Prevention and Control. Introduction to the annual epidemiological report. Q fever Annual Epide-miological Report. ECDC, Stockholm, 2019.
Eldin C, Mélenotte C, Mediannikov O, Ghigo E, Million M, Edouard S, Mege JL, Maurin M, Raoult D (2017) From Q fever to Coxiella burnetii infection: a paradigm change. Clin Microbiol Rev 30: 115-190
Enserink M (2010) Infectious diseases. Questions abound in Q-fever explosion in the Netherlands. Science 327: 266-267
Ghaoui H, Achour N, Saad-Djaballah A, Smai A, Temim S, Bitam I (2019) Between livestock’s and humans, Q fever disease is emerging at low noise. Acta Sci Microbiol 2: 104-132
Hubálek Z, Juricová Z, Svobodová S, Halouzka J (1993) A serologic survey for some bacterial and viral zoonoses in game animals in the Czech Republic. J Wildl Dis 29: 604-607.
Literák I (1990) Occurrence of Coxiella burnetii antibodies in cattle, sheep and small terrestrial mammals in the western region of Bohemia [Article in Czech]. Vet Med (Praha) 40: 77-80.
Literák I, Calvo Rodríguez B (1994) Latent Q fever in cattle in southern Moravia (Czech Republic). Cent Eur J Publ Health 2: 91-94.
Literák I, Kroupa L (1998) Herd-level Coxiella burnetii seroprevalence was not associated with herd-level bree- ding performance in Czech dairy herds. Prev Vet Med 33: 261-265.
Loftis AD, Reeves WK, Szumlas DE, Abbassy MM, Helmy IM, Moriarity JR (2006) Rickettsial agents in Egyptian ticks collected from domestic animals. Exp Appl Acarol 40: 67-81.
López-Gatius F, Almeria S, Garcia-Ispierto I (2012) Serolo- gical screening for Coxiella burnetii infection and related reproductive perfor-mance in high producing dairy cows. Res Vet Sci 93: 67-73.
Maurin M, Raoult D (1999) Q fever. Clin Microbiol Rev 12: 518-553.
Niemczuk K, Szymańska-Czerwińska M, Zarzecka A, Konarska H (2011) Q fever in a cattle herd and humans in the south-eastern Poland. Laboratory diagnosis of the disease using serological and molecular methods. Bull Vet Inst Pulawy 55: 593-598.
OIE (2018) Manual of diagnostic tests and vaccines for terrestrial animals. Volume 1, Part 3, Section 3. Chapter 3.1.16. Q fever. World Or-ganisation for Animal Health, 8th edition.
Patocka F, Kubelka V (1953) Studies on Q fever. Cesk Epidemiol Mikrobiol Immunol 2: 340-352.
R Core Team (2020) R: A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. URL https://www.R-project.org/
Řehácek J. (1987) Epidemiology and significance of Q fever in Czechoslovakia. Zbl Bakt Mikrobiol Hyg Ser A: Med Microbiol Infect Dis Virol Parasitol 267: 16-19.
Rodolakis A, Berri M, Hechard C, Caudron C, Souriau A, Bodier CC, Blanchard B, Camuset P, Devillechaise P, Natorp JC, Vadet JP, Arri-cau- Bouvery N (2007) Compa- rison of Coxiella burnetii shedding in milk of dairy bovine, caprine, and ovine herds. J Dairy Sci 90: 5352-5360.
Schimmer B, Schotten N, van Engelen E, Hautvast JLAP, Schneeberger M, van Duijnhoven YTHP (2014) Coxiella burnetii seroprevalence and risk for humans on dairy cattle farms, the Netherlands, 2010-2011. Emerg Infect Dis 20: 417-425.
Splino M, Beran J, Chlíbek R (2003) Q fever outbreak during the Czech Army deployment in Bosnia. Mil Med 168: 840-842.
Statni Veterinarni Sprava (2018) Official Veterinary Report of the Czech Republic [in Czech].
Szymańska-Czerwińska M, Galińska EM, Niemczuk K, Zasępa M (2013) Prevalence of Coxiella burnetii infection in foresters in the south-eastern Poland and comparison of diagnostic methods. Ann Agric Environ Med 20: 699-704.
van der Hoek W, Morroy G, Renders NH, Wever PC, Hermans MH, Leenders AC, Schneeberger PM (2012) Epidemic Q fever in humans in the Netherlands. Adv Exp Med Biol 984: 329-364.
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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

To develop a sensitive, specific, and rapid approach for the detection Getah virus (GETV), a set of primers targeting the conserved region of the E1 gene was created. The TaqMan-based real-time PCR method for GETV detection was developed by optimizing the reaction conditions. The method demonstrated excellent specificity, and amplification did not occur with the causative agents of all prevalent swine viral infections (CSFV, PRRSV, PRV, PEDV, PTV, and JEV), except GETV. Additionally, upon assessing the sensitivity of the method, the minimum detection limit for GETV was found to be 5.94 copies/μL, which is 10 times higher than that of the traditional PCR approach. Further, the intra- and inter-assay variation coefficients were less than 1%, demonstrating good repeatability. Moreover, GETV was found in 10 of the 20 field serum samples using real-time PCR but only in three of the samples using traditional PCR. Consequently, the first GETV TaqMan-based real-time PCR approach based on the E1 gene was developed for GETV pathogenic diagnoses, and this exhibited high specificity, sensitivity, and repeatability. This assay is practical for the pathogenic diagnosis and epidemiology of GETV.
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Authors and Affiliations

A. Lin
1
X. Hu
1
S. Cui
1
T. Yang
1
Z. Zhang
1
P. Li
1
M. Guo
1
Y. Lu
1

  1. College of Life Sciences and Resource Environment, Yichun University, No 576, Xuefu Road, Yuanzhou district, Yichun, Jiangxi, 336000, China
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Abstract

Diarrhea caused by parasitic agents is common in neonatal calves and leads to significant economic losses in cattle farms worldwide. Cryptosporidium spp. is one of the most frequently detected parasitic agents causing diarrhea in neonatal calves. The aim of this study was to investigate the presence of Cryptosporidium spp. on a dairy farm which a has major diarrhea problem. Samples were collected from calves, cows, drinking bowls, and two different artesian water sources, as well as from the environment. All fecal samples were investigated using Kinyoun acid-fast stained slides and real-time PCR targeting the Cryptosporidium spp. COWP gene. In addition, species identification was performed by nested PCR targeting the Cryptosporidium spp. COWP gene and sequencing. Cryptosporidium spp. was detected in 11 calves (30.55%; 11/36) by real-time PCR and the cows were negative. Among real-time PCR positive samples, only five were also found positive by microscopy. Moreover, Cryptosporidium spp. was found in one of the two artesian water sources and five environmental samples by real-time PCR. Among these positive samples, eight were sequenced. According to the RFLP pattern, BLAST and, phylogenetic analyses, all sequenced samples were Cryptosporidium parvum. These findings show the importance of C. parvum as a cause of calf diarrhea on dairy farms.
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Bibliography


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Doungmala P, Phuektes P, Taweenan W, Sangmaneedet S, Japa O (2019) Prevalence and species identification of Cryptosporidium spp. In the newborn dairy calves from Muang District, Khon Kaen Province, Thailand. Vet World 12: 1454-1459.
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Fayer R, Santín M, Macarisin D (2010) Cryptosporidium ubiquitum n. sp. in animals and humans. Vet Parasitol 172: 23-32.
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Authors and Affiliations

M. Karakavuk
1 2
H. Can
3
M. Döşkaya
1
T. Karakavuk
1
S. Erkunt-Alak
3
A.E. Köseoğlu
3
A. Gül
4
C. Ün
3
Y. Gürüz
1
A. Değirmenci-Döşkaya
1

  1. Ege University Faculty of Medicine, Department of Parasitology, Bornova, İzmir, Turkey
  2. Ege University, Ödemiş Vocational School, Veterinary technology programs, Ödemiş, Izmir, Turkey
  3. Ege University Faculty of Science, Department of Biology, Molecular Biology Section, Bornova, İzmir, Turkey
  4. Ege University Faculty of Engineering, Department of Bioengineering, Bornova, İzmir, Turkey
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Abstract

In this study, we developed a SYBR Green I real-time PCR method for the rapid and sensitive detection of novel porcine parvovirus 7 (PPV7). Specific primers were designed based on the highly conserved region within the Capsid gene of PPV7. The established method was 1,000 times more sensitive than the conventional PCR method and had a detection limit of 35.6 copies. This method was specific and had no cross-reactions with PCV2, PCV3, PRV, PEDV, PPV1, and PPV6. Experiments testing the intra and interassay precision demonstrated a high reproducibility. Testing the newly established method with 200 clinical samples revealed a detection rate up to 17.5% higher than that of the conventional PCR assay. The established method could provide technical support for clinical diagnosis and epidemiological investigation of PPV7.
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Authors and Affiliations

Y.D. Li
1
Z.D. Yu
2
C.X. Bai
2
D. Zhang
2
P. Sun
2
M.L Peng
2
H. Liu
3
ORCID: ORCID
J. Wang
4
Y. Wang
2
ORCID: ORCID

  1. Municipal Key Laboratory of Virology, Ningbo Municipal Center for Disease Control and Prevention, Ningbo 315010, PR China
  2. Anhui Province Key Laboratory of Veterinary Pathobiology and Disease Control, College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, PR China
  3. Anhui Animal Diseases Prevention and Control Center and Key Laboratory of Veterinary Pathobiology and Disease Prevention and Control of Anhui Province, Hefei 230091, PR China
  4. Animal Husbandry Base Teaching and Research Section, College of Animal Science and Technology, Hebei North University, Hebei 075000, PR China
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Abstract

The present study was aimed to establish a novel TaqMan real-time PCR (RTm-PCR) for detecting and typing bovine viral diarrhea virus (BVDV), and also to develop a diagnostic proto- col which simplifies sample collection and processing. Universal primers and TaqMan-MGB probes were designed from the known sequences of conserved 5′ - and 3′-untranslated regions (5’UTR, 3’UTR) of the NADL strain of BVDV. Prior to optimizing the assay, cDNAs were tran- scribed in vitro to make standard curves. The sensitivity, specificity and stability (reproducibility) were evaluated. The RTm-PCR was tested on the 312 feces specimens collected from persistently infected (PI) calves. The results showed the optimum conditions for RTm-PCR were 17.0 μmol/L primer, 7.5 μmol/L probe and 51.4°C annealing temperature. The established TaqMan RTm-PCR assay could specially detect BVDV without detecting any other viruses. Its detection limit was 1.55×100 copies/μL for viral RNA. It was 10000-fold higher than conventional PCR with excel- lent specificity and reproducibility. 312 samples were tested using this method and universal PCR from six dairy farms, respectively. Positive detections were found in 49 and 44 feces samples, respectively. The occurrence rate was 89.80%. In conclusion, the established TaqMan RTm-PCR could rapidly detect BVDV and effectively identify PI cattle. The detection limit of RTm-PCR was 1.55 copies/μL. It will be beneficial for enhancing diagnosis and therapy efficacy and reduce losses in cattle farms.

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Authors and Affiliations

H. Liang
J. Geng
S. Bai
A. Aimuguri
Z. Gong
R. Feng
X. Shen
S. Wei
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Abstract

The present study investigated the expression of androgen receptor (AR) in neurons of the anterior pelvic ganglion (APG) and celiac-superior mesenteric ganglion (CSMG; ganglion not involved in the innervation of reproductive organs) in the male pig with quantitative real-time PCR (qPCR) and immunohistochemistry. qPCR investigations revealed that the level of AR gene expression in the APG tissue was approximately 2.5 times higher in the adult (180-day-old) than in the juvenile (7-day-old) boars. Furthermore, in both the adult and juvenile animals it was sig- nificantly higher in the APG than in CSMG tissue (42 and 85 times higher, respectively). Immu- nofluorescence results fully confirmed those obtained with qPCR. In the adult boars, nearly all adrenergic (DβH-positive) and the majority of non-adrenergic neurons in APG stained for AR. In the juvenile animals, about half of the adrenergic and non-adrenergic neurons were AR-posi- tive. In both the adult and juvenile animals, only solitary CSMG neurons stained for AR. The present results suggest that in the male pig, pelvic neurons should be considered as an element of highly testosterone-dependent autonomic circuits involved in the regulation of urogenital func- tion, and that their sensitization to androgens is a dynamic process, increasing during the prepu- bertal period.

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Authors and Affiliations

J. Kaleczyc
N. Kasica-Jarosz
Z. Pidsudko
A. Przyborowska
W. Sienkiewicz

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