Presence of porcine cytomegalovirus, a porcine roseolovirus, in wild boars in Italy and Germany

Hansen, Sabrina; Menandro, Maria Luisa; Franzo, Giovanni; Krabben, Ludwig; Marino, Stephen F.; Kaufer, Benedikt B.; Denner, Joachim

doi:10.1007/s00705-022-05690-6

Cited by 4 publications

(8 citation statements)

References 63 publications

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“…When however, the antibody titer declined or is negative, testing of organ biopsies is recommended. In a schematic presentation based on the results presented in 19 and, 20 as well in previous publications 5, [15][16][17] studying methods and time line to detect PCMV/PRV, possible test strategies are indicated (Figure 3). The related human roseolovirus HHV-6 is better studied in this context.…”

Section: Discussionmentioning

confidence: 99%

“…F I G U R E 3 Schematic presentation of strategies for testing for PCMV/PRV using immunological methods detecting virus-specific antibodies and PCR-based methods in pigs of different age based on results published in Ref. [15][16][17][18][19][20]27 Therefore, also testing of organs was included. The broader the bars the more effective testing will be.…”

Section: Discussionmentioning

confidence: 99%

“…Halecker et al 19 used a Western blot assay, which was established in 2016 21 and since then used for screening of slaughterhouse pigs, Göttingen and Aachen minipigs 21 as well as wild boars in Italy and Germany. 15 Originally this assay used a recombinant N-terminal part (designated R1) and a C-terminal part (R2) of the glycoprotein B (gB) of PCMV/PRV 21 (Figure 2A). However, it was shown that R1…”

Section: Immunological Methodsmentioning

confidence: 99%

“…Detection of PCMV/PRV by PCR in adult animals will be observed if the virus is reactivated by stress, immune reaction or other external factors. 15 Reactivation of the virus in older animals will at the same time trigger the immune response. PCR is also required to exclude infection of the animal in adulthood.…”

Section: Recommendationsmentioning

confidence: 99%

“…This usually occurs in freshly infected young piglets or during sporadic reactivation of the virus in adult pigs. 15 In one study nasal shedding of the virus was observed between days 14 and 28 after infection. 16 Usually pigs are infected from the mother and PCMV/PRV is found during acute infection in the tissues throughout the body including the nose of newly infected newborn piglets where it causes rhinitis and conjunctivitis.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of methods for the detection of porcine cytomegalovirus/roseolovirus in relation to biosafety monitoring of xenotransplantation products

Denner,

Jhelum,

Hansen

et al. 2023

Xenotransplantation

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BackgroundThe porcine cytomegalovirus, a porcine roseolovirus (PCMV/PRV), is widely distributed in pig populations. It has been shown that PCMV/PRV was transmitted by pig xenotransplants to non‐human primates, and significantly reduced the survival time of the recipient. PCMV/PRV was also transmitted during the first transplantation of a pig heart into a human patient. PCMV/PRV establishes a lifelong persistent infection (latency) in the host, is difficult to detect in this stage, and consequential poses a threat to future clinical xenotransplantations. Therefore, sensitive and specific methods and goal‐oriented strategies how, when, and where to test should be used for screening donor pigs.MethodsIn this study we compared experimentally the PCMV/PRV detection methods including PCR‐based (real‐time PCR, nested PCR) and immunological methods (Western blot assay, ELISA) recently published by Halecker et al. (Sci. Rep. 2022;12(1):21545) and Fischer et al. (Xenotransplantation 2023:e12803). We also compared the antigens used for antibody detection (a recombinant protein and synthetic peptides corresponding to a conserved region of the glycoprotein B, gB).ResultsThe published methods can be used for screening donor pigs, with the results being similar. The antigens used for the detection of PCMV/PRV‐specific antibodies are almost identical and give comparable results. Overall, the optimal diagnostic tests, the samples used for testing and the time of sampling play a crucial role in preventing the transmission of PCMV/PRV during xenotransplantation.ConclusionSensitive methods are available to screen donor pigs for PCMV/PRV, but a rational application of a combination of PCR‐based and immunological methods as well as rational detection strategies are important for the detection of the virus during latency.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Immunological Methodsmentioning

confidence: 99%

Section: Recommendationsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Comparison of methods for the detection of porcine cytomegalovirus/roseolovirus in relation to biosafety monitoring of xenotransplantation products

Denner,

Jhelum,

Hansen

et al. 2023

Xenotransplantation

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First virological and pathological study of Göttingen Minipigs with Dippity Pig Syndrome (DPS)

Jhelum

Grand²,

Jacobsen³

et al. 2023

Preprint

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Dippity Pig Syndrome (DPS) is a well-known but rare complex of clinical signs affecting minipigs, which has not been thoroughly investigated yet. Clinically affected animals show acute appearance of red, exudating lesions across the spine. The lesions are painful, evidenced by arching of the back (dipping), and the onset of clinical symptoms is generally sudden. In order to understand the pathogenesis, histological and virological investigations were performed in affected and unaffected Göttingen Minipigs (GöMPs). The following DNA viruses were screened for using PCR-based methods: Porcine cytomegalovirus (PCMV), which is a porcine roseolovirus (PCMV/PRV), porcine lymphotropic herpesviruses (PLHV-1, PLHV-2, PLHV-3), porcine circoviruses (PCV1, PCV2, PCV3, PCV4), porcine parvovirus 1 (PPV1), and Torque Teno sus virus (TTSuV1, TTSuV2). Screening was also performed for integrated porcine endogenous retroviruses (PERV-A, PERV-B, PERV-C) and recombinant PERV-A/C and their expression as well as for the RNA viruses hepatitis E virus (HEV) and SARS-CoV-2. Eight clinically affected and one unaffected GöMPs were analyzed. Additional unaffected minipigs had been analyzed in the past. The analyzed GöMPs contained PERV-A and PERV-B integrated in the genome, which are present in all pigs and PERV-C, which is present in most, but not all pigs. In one affected GöMPs recombinant PERV-A/C was detected in blood. In this animal a very high expression of PERV mRNA was observed. PCMV/PRV was found in three affected animals, PCV1 was found in three animals with DPS and in the healthy minipig, and PCV3 was detected in two animals with DPS and in the unaffected minipig. Most importantly, in one animal only PLHV-3 was detected. It was found in the affected and unaffected skin, and in other organs. Unfortunately, PLHV-3 could not be studied in all other affected minipigs. None of the other viruses were detected and using electron microscopy, no virus particles were found in the affected skin. This data identified some virus infections in GöMPs with DPS and assign a special role to PLHV-3. Since PCMV/PRV, PCV1, PCV3 and PLHV-3 were also found in unaffected animals, a multifactorial cause of DPS is suggested. However, elimination of the viruses from GöMPs may prevent DPS.

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Detection of porcine cytomegalovirus, a roseolovirus, in pig ovaries and follicular fluid: implications for somatic cells nuclear transfer, cloning and xenotransplantation

Hansen

Fischer

Krabben

et al. 2023

Virol J

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Background Porcine cytomegalovirus (PCMV) is a porcine roseolovirus (PCMV/PRV) which is widely distributed in pigs. Transmission of PCMV/PRV in preclinical xenotransplantations was shown to significantly reduce the survival time of the pig transplants in non-human primates. PCMV/PRV was also transmitted in the first transplantation of a pig heart into a human patient. To analyze how PCMV/PRV could be introduced into pig breeds, especially considering cloned transgenic pigs, and subsequently spread in breeding facilities, we screened ovaries and derived materials which are used to perform somatic cell nuclear transfer (SCNT). Methods DNA was isolated from ovarian tissues, follicular fluids, oocytes with cumulus cells, denuded oocytes and parthenotes. A real-time PCR with PCMV/PRV-specific primers and a probe was performed to detect PCMV/PRV. Furthermore, a Western blot assay using a recombinant fragment of the gB protein of PCMV/PRV was performed to screen for virus-specific antibodies in the follicular fluids. Results PCMV/PRV was found by real-time PCR in ovarian tissues, in the follicular fluid and in oocytes. In parthenotes the virus could not be detected, most-likely due to the low amount of DNA used. By Western blot assay specific antibodies against PCMV/PRV were found in 19 of 20 analyzed follicular fluids. Conclusion PCMV/PRV was found in ovarian tissues, in the follicular fluids and also in denuded oocytes, indicating that the virus is present in the animals of which the oocytes were taken from. Despite several washing steps of the denuded oocytes, which are subsequently used for microinjection or SCNT, the virus could still be detected. Therefore, the virus could infect oocytes during genetic modifications or stay attached to the surface of the oocytes, potentially infecting SCNT recipient animals.

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Presence of porcine cytomegalovirus, a porcine roseolovirus, in wild boars in Italy and Germany

Cited by 4 publications

References 63 publications

Comparison of methods for the detection of porcine cytomegalovirus/roseolovirus in relation to biosafety monitoring of xenotransplantation products

Comparison of methods for the detection of porcine cytomegalovirus/roseolovirus in relation to biosafety monitoring of xenotransplantation products

First virological and pathological study of Göttingen Minipigs with Dippity Pig Syndrome (DPS)

Detection of porcine cytomegalovirus, a roseolovirus, in pig ovaries and follicular fluid: implications for somatic cells nuclear transfer, cloning and xenotransplantation

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