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Koi herpesvirus (KHV) is a highly contagious virus that causes KHV disease (KHVD) inducing high mortality in carp and koi (Cyprinus carpio L.). In the late stage, latency occurs with very low, often non‐detectable virus concentrations, which represents a challenge for virus detection. After validation according to OIE recommendations, an antibody ELISA was established to recognize antibodies of C. carpio against KHV infection. In this study, the ELISA was modified to detect anti‐KHV antibodies from a non‐cyprinid fish. Experimentally infected rainbow trout (Oncorhynchus mykiss) were able to transmit KHV to naïve carp at two different temperatures, demonstrating their potential as a reservoir host. At 20°C, KHVD was induced in carp but not at 15°C. Unexpectedly, rainbow trout developed humoral response against KHV at both temperatures. In contrast to carp, at 15°C trout produced neutralizing antibodies but not at 20°C. While antibodies obtained from infected carp sera reacted in a similar way against all KHV, antibodies from rainbow trout sera reacted differently to the same isolates by ELISA. The data show that even when non‐cyprinid fish species are infected with KHV, they can produce antibodies that differ from those observed in carp.
Sven Michael Bergmann; Juliane Dabels; Sandro Klafack; Yeonwha Jin; Pei‐Yu Lee; Arndt Christian Hofmann; Yingying Wang; Qing Wang; Yingying Li; Weiwei Zeng; Angela Lusiastuti; Shucheng Zheng; Yijuan Jin; Jolanta Kiełpińska; Sean Monaghan. Serological responses to koi herpesvirus (KHV) in a non‐cyprinid reservoir host. Journal of Fish Diseases 2021, 1 .
AMA StyleSven Michael Bergmann, Juliane Dabels, Sandro Klafack, Yeonwha Jin, Pei‐Yu Lee, Arndt Christian Hofmann, Yingying Wang, Qing Wang, Yingying Li, Weiwei Zeng, Angela Lusiastuti, Shucheng Zheng, Yijuan Jin, Jolanta Kiełpińska, Sean Monaghan. Serological responses to koi herpesvirus (KHV) in a non‐cyprinid reservoir host. Journal of Fish Diseases. 2021; ():1.
Chicago/Turabian StyleSven Michael Bergmann; Juliane Dabels; Sandro Klafack; Yeonwha Jin; Pei‐Yu Lee; Arndt Christian Hofmann; Yingying Wang; Qing Wang; Yingying Li; Weiwei Zeng; Angela Lusiastuti; Shucheng Zheng; Yijuan Jin; Jolanta Kiełpińska; Sean Monaghan. 2021. "Serological responses to koi herpesvirus (KHV) in a non‐cyprinid reservoir host." Journal of Fish Diseases , no. : 1.
The hemorrhagic disease of grass carp (HDGC) caused by grass carp reovirus (GCRV) still poses a great threat to the grass carp industry. Isolation and identification of the GCRV genotype I (GCRV-I) has been rarely reported in the past decade. In this study, a new GCRV was isolated from diseased fish with severe symptoms of enteritis and mild hemorrhages on the body surface. The isolate was further identified by cell culture, transmission electron, indirect immunofluorescence, and SDS-PAGE electrophoretic pattern analysis of genomic RNA. The results were consistent with the new isolate as a GCRV-I member and tentatively named GCRV-GZ1208. Both grass carp and rare minnow infected by the GCRV-GZ1208 have no obvious hemorrhagic symptoms, and the final mortality rate was ≤10%, indicating that it may be a low virulent isolate. GZ1208 possessed highest genomic homology to 873/GCHV (GCRV-I) and golden shiner reovirus (GSRV). Additionally, it was found a 90.7–98.3% nucleotide identity, a 96.4–100% amino acid identity, and <50% identity with GCRV-II and III genotypes. Interestingly, the sequences of some segments of GZ1208 were similar to GCRV-8733/GCHV, whereas the remaining segments were more closely related to GSRV, suggesting that a recombination event had occurred. Bootscan analysis of the complete genomic sequence confirmed this hypothesis, and recombination events between 873/GCHV and other GSRV-like viruses were also accompanied by gene mutations.
Weiwei Zeng; Sven Bergmannc; Hanxu Dong; Ying Yang; Minglin Wu; Hong Liu; Yanfeng Chen; Hua Li. Identification, Virulence, and Molecular Characterization of a Recombinant Isolate of Grass Carp Reovirus Genotype I. Viruses 2021, 13, 807 .
AMA StyleWeiwei Zeng, Sven Bergmannc, Hanxu Dong, Ying Yang, Minglin Wu, Hong Liu, Yanfeng Chen, Hua Li. Identification, Virulence, and Molecular Characterization of a Recombinant Isolate of Grass Carp Reovirus Genotype I. Viruses. 2021; 13 (5):807.
Chicago/Turabian StyleWeiwei Zeng; Sven Bergmannc; Hanxu Dong; Ying Yang; Minglin Wu; Hong Liu; Yanfeng Chen; Hua Li. 2021. "Identification, Virulence, and Molecular Characterization of a Recombinant Isolate of Grass Carp Reovirus Genotype I." Viruses 13, no. 5: 807.
Koi herpesvirus disease (KHVD) caused by the koi herpesvirus (KHV) is difficult to diagnose in live fish, presenting a challenge to the koi industry. The enzyme‐linked immunosorbent assay (ELISA) method cannot be widely used to detect KHV because few commercial anti‐KHV antibody exists. Here, we developed an anti‐ORF132 polyclonal antibody and confirmed its reactivity via indirect immunofluorescence assay and Western blotting. A double‐antibody sandwich ELISA (DAS‐ELISA) was established to detect KHV, monoclonal antibody 1B71B4 against ORF92 was used as the capture antibody, and the detection antibody was the polyclonal antibody against the truncated ORF132. The lowest limit was 1.56 ng/ml KHV. Furthermore, the DAS‐ELISA reacted with KHV isolates, while no cross‐reactions occurred with carp oedema virus, spring viraemia of carp virus, frog virus 3 and grass carp reovirus. Two hundred koi serum samples from Guangdong, China, were used in the DAS‐ELISA test, and the positive rate of the koi sera was 13%. The clinical sensitivity and specificity of the DAS‐ELISA relative to the traditional PCR method were 66.7% and 97.6%, respectively. Our findings may be useful for diagnosing and preventing KHVD in koi and common carp.
Yingying Li; Qing Wang; Feng Hu; Yingying Wang; Sven M. Bergmann; Weiwei Zeng; Jiyuan Yin; Cunbin Shi. Development of a double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA) for the detection of KHV. Journal of Fish Diseases 2021, 44, 913 -921.
AMA StyleYingying Li, Qing Wang, Feng Hu, Yingying Wang, Sven M. Bergmann, Weiwei Zeng, Jiyuan Yin, Cunbin Shi. Development of a double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA) for the detection of KHV. Journal of Fish Diseases. 2021; 44 (7):913-921.
Chicago/Turabian StyleYingying Li; Qing Wang; Feng Hu; Yingying Wang; Sven M. Bergmann; Weiwei Zeng; Jiyuan Yin; Cunbin Shi. 2021. "Development of a double‐antibody sandwich enzyme‐linked immunosorbent assay (DAS‐ELISA) for the detection of KHV." Journal of Fish Diseases 44, no. 7: 913-921.
Tilapia lake virus (TiLV) is a newly emerging pathogen responsible for high mortality and economic losses in the global tilapia industry. Currently, no antiviral therapy or vaccines are available for the control of this disease. The goal of the present study was to evaluate the immunological effects and protective efficacy of formaldehyde- and β-propiolactone-inactivated vaccines against TiLV in the presence and absence of the Montanide IMS 1312 VG adjuvant in tilapia. We found that β-propiolactone inactivation of viral particles generated a vaccine with a higher protection efficacy against virus challenge than did formaldehyde. The relative percent survivals of vaccinated fish at doses of 108, 107, and 106 50% tissue culture infectious dose (TCID50)/mL were 42.9%, 28.5%, and 14.3% in the absence of the adjuvant and 85.7%, 64.3%, and 32.1% in its presence, respectively. The vaccine generated specific IgM and neutralizing antibodies against TiLV at 3 weeks following immunization that were significantly increased after a second booster immunization. The steady state mRNA levels of the genes tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interferon γ (IFN-γ), cluster of differentiation 4 (CD4), major histocompatibility complex (MHC)-Ia, and MHC-II were all increased and indicated successful immune stimulation against TiLV. The vaccine also significantly lowered the viral loads and resulted in significant increases in survival, indicating that the vaccine may also inhibit viral proliferation as well as stimulate a protective antibody response. The β-propiolactone-inactivated TiLV vaccine coupled with the adjuvant Montanide IMS 1312 VG and booster immunizations can provide a high level of protection from virus challenge in tilapia.
Weiwei Zeng; Yingying Wang; Huzi Hu; Qing Wang; Sven Bergmann; Yahui Wang; Bo Li; Yuefeng Lv; Hua Li; Jiyuan Yin; Yingying Li. Cell Culture-Derived Tilapia Lake Virus-Inactivated Vaccine Containing Montanide Adjuvant Provides High Protection against Viral Challenge for Tilapia. Vaccines 2021, 9, 86 .
AMA StyleWeiwei Zeng, Yingying Wang, Huzi Hu, Qing Wang, Sven Bergmann, Yahui Wang, Bo Li, Yuefeng Lv, Hua Li, Jiyuan Yin, Yingying Li. Cell Culture-Derived Tilapia Lake Virus-Inactivated Vaccine Containing Montanide Adjuvant Provides High Protection against Viral Challenge for Tilapia. Vaccines. 2021; 9 (2):86.
Chicago/Turabian StyleWeiwei Zeng; Yingying Wang; Huzi Hu; Qing Wang; Sven Bergmann; Yahui Wang; Bo Li; Yuefeng Lv; Hua Li; Jiyuan Yin; Yingying Li. 2021. "Cell Culture-Derived Tilapia Lake Virus-Inactivated Vaccine Containing Montanide Adjuvant Provides High Protection against Viral Challenge for Tilapia." Vaccines 9, no. 2: 86.
Grass carp reovirus (GCRV) leads to severe hemorrhagic disease in grass carp (Ctenopharyngodon idella) and causes economic losses in grass carp aquaculture. Recent epidemiological investigations showed that GCRV genotype II is the dominant subtype in China. Therefore, it is very important to develop a novel vaccine for preventing diseases caused by GCRV genotype II. In this study, we employed a bac-to-bac expression system to generate GCRV-II-based virus-like particles (VLPs). Previous studies have shown that the structural proteins VP3, VP4, and VP38 encoded by the segments S3, S6, and S10 of type II GCRV are immunogenic. Hence, the GCRV-VLPs were produced by co-infection of sf9 cells with recombinant baculoviruses PFBH-VP3, PFBH-VP4, and PFBH-VP38. The expressions of VP3, VP4, and VP38 proteins in GCRV-VLPs were tested by IFA and Western blot analysis. By electron microscopic observations of ultrathin sections, purified VLPs showed that the expressed proteins are similar in shape to GCRV genotype II with a size range from 40 nm to 60 nm. The immunogenicity of GCRV-VLPs was evaluated by the injection immunization of grass carp. The analysis of serum-specific IgM antibody showed that grass carp immunized with GCRV-VLPs produced GCRV-specific antibodies. Furthermore, injection with GCRV-VLPs increased the expressions of immune-related genes (IgM, IFN, TLR3, TLR7) in the spleen and kidney. In addition, grass carp immunized with a GCRV-VLPs-based vaccine showed a relative percent survival rate (RPS) of 83.33% after challenge. The data in this study showed that GCRV-VLPs demonstrated an excellent immunogenicity and represent a promising approach for vaccine development against GCRV genotype II infection.
Ting Gao; Caixia Gao; Siyu Wu; Yingying Wang; Jiyuan Yin; Yingying Li; Weiwei Zeng; Sven Bergmann; Qing Wang. Recombinant Baculovirus-Produced Grass Carp Reovirus Virus-Like Particles as Vaccine Candidate that Provides Protective Immunity against GCRV Genotype II Infection in Grass Carp. Vaccines 2021, 9, 53 .
AMA StyleTing Gao, Caixia Gao, Siyu Wu, Yingying Wang, Jiyuan Yin, Yingying Li, Weiwei Zeng, Sven Bergmann, Qing Wang. Recombinant Baculovirus-Produced Grass Carp Reovirus Virus-Like Particles as Vaccine Candidate that Provides Protective Immunity against GCRV Genotype II Infection in Grass Carp. Vaccines. 2021; 9 (1):53.
Chicago/Turabian StyleTing Gao; Caixia Gao; Siyu Wu; Yingying Wang; Jiyuan Yin; Yingying Li; Weiwei Zeng; Sven Bergmann; Qing Wang. 2021. "Recombinant Baculovirus-Produced Grass Carp Reovirus Virus-Like Particles as Vaccine Candidate that Provides Protective Immunity against GCRV Genotype II Infection in Grass Carp." Vaccines 9, no. 1: 53.
Rainbow trout (Oncorhynchus mykiss) and common carp (Cyprinus carpio) are the two most common species in traditional fish farming in Germany. Their aquaculture is threatened upon others by viruses that can cause a high mortality. Therefore, this work focuses on three viruses—viral haemorrhagic septicaemia virus, infectious hematopoietic necrosis virus and cyprinid herpesvirus 3 (CyHV‐3)—that endanger these species. To prevent their spread and contain further outbreaks, it is essential to know how long they can outlast in environmental waters and what affects their infectivity outside the host. Hence, the stability of the target viruses in various water matrices was examined and compared in this work. In general, all three viruses were quite stable within sterile water samples (showing mostly ≤1 log reduction after 96 hr) but were inactivated faster and to a higher extent (up to five log steps within 96 hr) in unsterile environmental water samples. The inactivation of the viruses correlated well with the increasing bacterial load of the samples, suggesting that bacteria had the greatest effect on their stability in the examined samples. In comparison, CyHV‐3 seemed to be the most sensitive and maintained its infectivity for the shortest period.
Johanna Ullrich; Jürgen Christian; Sven Michael Bergmann; Martin Oberle; Anna Maria Becker. Stability of viral haemorrhagic septicaemia virus, infectious hematopoietic necrosis virus and cyprinid herpesvirus 3 in various water samples. Journal of Fish Diseases 2020, 44, 379 -390.
AMA StyleJohanna Ullrich, Jürgen Christian, Sven Michael Bergmann, Martin Oberle, Anna Maria Becker. Stability of viral haemorrhagic septicaemia virus, infectious hematopoietic necrosis virus and cyprinid herpesvirus 3 in various water samples. Journal of Fish Diseases. 2020; 44 (4):379-390.
Chicago/Turabian StyleJohanna Ullrich; Jürgen Christian; Sven Michael Bergmann; Martin Oberle; Anna Maria Becker. 2020. "Stability of viral haemorrhagic septicaemia virus, infectious hematopoietic necrosis virus and cyprinid herpesvirus 3 in various water samples." Journal of Fish Diseases 44, no. 4: 379-390.
Over the last years, there has been an enormous increase in the knowledge on koi herpesvirus (KHV), koi herpesvirus disease (KHVD), pathogenesis and virus variants. Different KHV lineages have clearly been identified, possible genomic changes during replication in different cell cultures at different temperatures but also in several hosts have been identified, a persistent stage of infection has been specified and it has been shown that infection with KHV is not host specific at all, but KHVD is. Additionally, it has been shown that it is possible to combat KHVD by immunization with inactivated and attenuated live vaccines using different delivery systems but also to benefit from alternative treatments with e.g. exopolysaccharids obtained from Arthrospira platensis.
Sven M. Bergmann; Yeonhwa Jin; Kati Franzke; Bianka Grunow; Q. Wang; Sandro Klafack. Koi herpesvirus (KHV) and KHV disease (KHVD) – a recently updated overview. Journal of Applied Microbiology 2020, 129, 98 -103.
AMA StyleSven M. Bergmann, Yeonhwa Jin, Kati Franzke, Bianka Grunow, Q. Wang, Sandro Klafack. Koi herpesvirus (KHV) and KHV disease (KHVD) – a recently updated overview. Journal of Applied Microbiology. 2020; 129 (1):98-103.
Chicago/Turabian StyleSven M. Bergmann; Yeonhwa Jin; Kati Franzke; Bianka Grunow; Q. Wang; Sandro Klafack. 2020. "Koi herpesvirus (KHV) and KHV disease (KHVD) – a recently updated overview." Journal of Applied Microbiology 129, no. 1: 98-103.
A brain cell line (CAMB) derived from hybrid snakehead (Channa argus (♂) × Channa maculata (♀)) was established by trypsin and collagenase combined digestion. The culturing conditions and cell biological characteristics were systematically studied. For growth of the cells, M199 medium containing 10% fetal bovine serum was used and at 27 °C incubated. Based on morphological analysis, CAMB cells were confirmed to be epithelial. The cell line has been subcultured more than 80 times since its initial primary culture. Chromosome analysis revealed that CAMB cells had an abnormal chromosome number 2n = 64, whereas the chromosome number in the hybrid snakehead was 45. The suitability of CAMB for tilapia lake virus (TiLV) was demonstrated. A CPE was observed after infection with TiLV-2017A. The highest TiLV titer was observed after 12 days post infection (dpi) and reached 107.2 TCID50/mL. The virus replication was confirmed by electron microscopic observations. Additionally, immunofluorescence assay confirmed the presence of TiLV-2017A after infection of CAMB. Therefore, CAMB cells can be a useful tool for the investigation of the pathogenesis of the TiLV induced disease in tilapia.
Yingying Wang; Zhili Li; Qing Wang; Weiwei Zeng; Yingying Li; Jiyuan Yin; Sven M. Bergmann; Xinping Zhu. Establishment of a brain cell line obtained from hybrids of Channa argus ×Channa maculata for the detection of tilapia lake virus. Microbial Pathogenesis 2019, 138, 103810 .
AMA StyleYingying Wang, Zhili Li, Qing Wang, Weiwei Zeng, Yingying Li, Jiyuan Yin, Sven M. Bergmann, Xinping Zhu. Establishment of a brain cell line obtained from hybrids of Channa argus ×Channa maculata for the detection of tilapia lake virus. Microbial Pathogenesis. 2019; 138 ():103810.
Chicago/Turabian StyleYingying Wang; Zhili Li; Qing Wang; Weiwei Zeng; Yingying Li; Jiyuan Yin; Sven M. Bergmann; Xinping Zhu. 2019. "Establishment of a brain cell line obtained from hybrids of Channa argus ×Channa maculata for the detection of tilapia lake virus." Microbial Pathogenesis 138, no. : 103810.
Viruses are able to evolve in vitro by mutations after serial passages in cell cultures, which can lead to either a loss, or an increase, of virulence. Cyprinid herpesvirus 3 (CyHV-3), a 295-kb double-stranded DNA virus, is the etiological agent of the koi herpesvirus disease (KHVD). To assess the influence of serial passages, an isolate of CyHV-3 (KHV-T) was passaged 99 times onto common carp brain (CCB) cells, and virus virulence was evaluated during passages through the experimental infections of common carp. After 78 CCB passages, the isolate was much less virulent than the original form. A comparative genomic analysis of these three forms of KHV-T (P0, P78 and P99) revealed a limited number of variations. The largest one was a deletion of 1363 bp in the predicted ORF150, which was detected in P78, but not in P99. This unexpected finding was confirmed by conventional PCR and digital PCR. The results presented here primarily suggest that, CyHV-3 evolves, at least in vitro, through an assemblage of haplotypes that alternatively become dominant or under-represented.
Sandro Klafack; Anna-Sophie Fiston-Lavier; Sven M. Bergmann; Saliha Hammoumi; Lars Schröder; Walter Fuchs; Angela Lusiastuti; Pei-Yu Lee; Sarahi Vega Heredia; Anne-Sophie Gosselin-Grenet; Jean-Christophe Avarre; Master student consortium; Master student consortium Master student consortium. Cyprinid herpesvirus 3 Evolves In Vitro through an Assemblage of Haplotypes that Alternatively Become Dominant or Under-Represented. Viruses 2019, 11, 754 .
AMA StyleSandro Klafack, Anna-Sophie Fiston-Lavier, Sven M. Bergmann, Saliha Hammoumi, Lars Schröder, Walter Fuchs, Angela Lusiastuti, Pei-Yu Lee, Sarahi Vega Heredia, Anne-Sophie Gosselin-Grenet, Jean-Christophe Avarre, Master student consortium, Master student consortium Master student consortium. Cyprinid herpesvirus 3 Evolves In Vitro through an Assemblage of Haplotypes that Alternatively Become Dominant or Under-Represented. Viruses. 2019; 11 (8):754.
Chicago/Turabian StyleSandro Klafack; Anna-Sophie Fiston-Lavier; Sven M. Bergmann; Saliha Hammoumi; Lars Schröder; Walter Fuchs; Angela Lusiastuti; Pei-Yu Lee; Sarahi Vega Heredia; Anne-Sophie Gosselin-Grenet; Jean-Christophe Avarre; Master student consortium; Master student consortium Master student consortium. 2019. "Cyprinid herpesvirus 3 Evolves In Vitro through an Assemblage of Haplotypes that Alternatively Become Dominant or Under-Represented." Viruses 11, no. 8: 754.
Recently, substantial mortality of farmed and wild tilapia caused by tilapia lake virus (TiLV) infection has been observed worldwide. However, sensitive and reliable diagnostic method is limited. A reverse transcription–loopmediated isothermal amplification (RT‐LAMP) assay has been applied for the detection of TiLV nucleotide sequence. Six primers targeting two locations on the target gene based on a highly conserved sequence in the segment 1 (S1) region of the TiLV genome have been designed. The optimized RT‐LAMP reaction was maintained at the isothermal condition of 63°C for 45 min. And the amplifications could be verified by turbidity or a colour change with the addition of SYBR Green I. Subsequently, RT‐LAMP products could be observed by a ladder pattern following gel electrophoresis. The species‐specific assay showed that the method was sensitive enough to detect as low as 1.6 copies of viral particle, and the assay was highly specific because no cross‐reactivity was observed with other pathogens, and had a diagnostic sensitivity and specificity of 100% when TiLV‐positive samples and non‐target virus were tested. In summary, all the results demonstrate that this RT‐LAMP is a rapid, effective and sensitive method for TiLV detection in tilapia aquaculture.
Jiyuan Yin; Qing Wang; Yingying Wang; Yingying Li; Weiwei Zeng; Jiexing Wu; Yan Ren; Yafang Tang; Caixia Gao; Huzi Hu; Sven M. Bergmann. Development of a simple and rapid reverse transcription–loopmediated isothermal amplification (RT‐LAMP) assay for sensitive detection of tilapia lake virus. Journal of Fish Diseases 2019, 42, 817 -824.
AMA StyleJiyuan Yin, Qing Wang, Yingying Wang, Yingying Li, Weiwei Zeng, Jiexing Wu, Yan Ren, Yafang Tang, Caixia Gao, Huzi Hu, Sven M. Bergmann. Development of a simple and rapid reverse transcription–loopmediated isothermal amplification (RT‐LAMP) assay for sensitive detection of tilapia lake virus. Journal of Fish Diseases. 2019; 42 (6):817-824.
Chicago/Turabian StyleJiyuan Yin; Qing Wang; Yingying Wang; Yingying Li; Weiwei Zeng; Jiexing Wu; Yan Ren; Yafang Tang; Caixia Gao; Huzi Hu; Sven M. Bergmann. 2019. "Development of a simple and rapid reverse transcription–loopmediated isothermal amplification (RT‐LAMP) assay for sensitive detection of tilapia lake virus." Journal of Fish Diseases 42, no. 6: 817-824.
Tilapia lake virus (TiLV) is an emerging disease threatening tilapia culture in many parts of the world. A cell line from the brain of tilapia, which was named TiB, was established, characterized and subcultured with more than 100 passages. The TiB cell line was optimally maintained at 27°C using medium 199 (M199) supplemented with 10% foetal bovine serum (FBS). Chromosome analysis revealed that 60% of TiB cells at passage 5 maintained the modal chromosome number 2n = 44, while at passage 60, there were 43% of TiB cells with the diploid chromosome number 2n = 50. A significant cytopathic effect was observed in TiB cells after infection with tilapia lake virus (TiLV‐2017A), and the viral replication in the cells was confirmed by transmission electron microscopy, immunofluorescence assays and viral titres, indicating the susceptibility of TiB cells to TiLV‐2017A. The viral titres of TiLV‐2017A in TiB cells reached 107.43 TCID50/ml within 10 days. The stable growth and susceptibility to fish viruses make TiB cells a useful tool for fish virus–host cell interaction and for immune response of fish.
Yingying Wang; Qing Wang; Weiwei Zeng; Jiyuan Yin; Yingying Li; Yan Ren; Cunbin Shi; Sven M. Bergmann; Xinping Zhu. Establishment and characterization of a cell line from tilapia brain for detection of tilapia lake virus. Journal of Fish Diseases 2018, 41, 1803 -1809.
AMA StyleYingying Wang, Qing Wang, Weiwei Zeng, Jiyuan Yin, Yingying Li, Yan Ren, Cunbin Shi, Sven M. Bergmann, Xinping Zhu. Establishment and characterization of a cell line from tilapia brain for detection of tilapia lake virus. Journal of Fish Diseases. 2018; 41 (12):1803-1809.
Chicago/Turabian StyleYingying Wang; Qing Wang; Weiwei Zeng; Jiyuan Yin; Yingying Li; Yan Ren; Cunbin Shi; Sven M. Bergmann; Xinping Zhu. 2018. "Establishment and characterization of a cell line from tilapia brain for detection of tilapia lake virus." Journal of Fish Diseases 41, no. 12: 1803-1809.
A disease with white spots in internal organs of Nile tilapia occurred in Zhanjiang, southern China. Multiple, white nodules, 0.8–2.2 mm in diameter, were scattered throughout the liver, spleen and kidney of diseased fish. Signs of nodules reproduced after artificial infection with the isolated strain. Isolated bacteria were Gram‐negative, facultative anaerobic, motile, short rod‐shaped, with a length of 1.2–2.2 μm. Morphological and biochemical tests, as well as phylogenetic analysis, all strongly indicated that the isolate from tilapia is identical to Aeromonas schubertii (A. schubertii) which temporary named LF1708 strain. Antibiotic sensitivity assays showed the LF1708 is sensitive to 24 of 27 tested antibiotics. Pathogenicity test revealed that the isolate at the dose of 3.75 × 106 CFU/g killed 100% of experimental tilapia within 2 days and the dose of 1 × 107 CFU/g killed 100% of experimental zebrafish within 1 day. Histopathology of diseased tilapia infected with A. schubertii showed numerous necrotic lesions widely distributed in spleen, liver and kidney, and infiltration with a large number of bacteria. To our knowledge, this was the first report that associated A. schubertii with mortality in tilapia.
C Liu; O Q Chang; D F Zhang; K B Li; F Wang; M H Lin; C B Shi; L Jiang; Q Wang; S M Bergmann. Aeromonas shuberti as a cause of multi-organ necrosis in internal organs of Nile tilapia,Oreochromis niloticus. Journal of Fish Diseases 2018, 41, 1529 -1538.
AMA StyleC Liu, O Q Chang, D F Zhang, K B Li, F Wang, M H Lin, C B Shi, L Jiang, Q Wang, S M Bergmann. Aeromonas shuberti as a cause of multi-organ necrosis in internal organs of Nile tilapia,Oreochromis niloticus. Journal of Fish Diseases. 2018; 41 (10):1529-1538.
Chicago/Turabian StyleC Liu; O Q Chang; D F Zhang; K B Li; F Wang; M H Lin; C B Shi; L Jiang; Q Wang; S M Bergmann. 2018. "Aeromonas shuberti as a cause of multi-organ necrosis in internal organs of Nile tilapia,Oreochromis niloticus." Journal of Fish Diseases 41, no. 10: 1529-1538.
Marine and aquaculture industries are important sectors of the food production and global trade. Unfortunately, the fish food industry is challenged with a plethora of infectious pathogens. The freshwater and marine fish communities are rapidly incorporating novel and most up to date techniques for detection, characterization and treatment strategies. Rapid detection of infectious diseases is important in preventing large disease outbreaks. One hundred forty-six articles including reviews papers were analyzed and their conclusions evaluated in the present paper. This allowed us to describe the most recent development research regarding the control of diseases in the aquatic environment as well as promising avenues that may result in beneficial developments. For the characterization of diseases, traditional sequencing and histological based methods have been augmented with transcriptional and proteomic studies. Recent studies have demonstrated that transcriptional based approaches using qPCR are often synergistic to expression based studies that rely on proteomic-based techniques to better understand pathogen-host interactions. Preventative therapies that rely on prophylactics such as vaccination with protein antigens or attenuated viruses are not always feasible and therefore, the development of therapies based on small nucleotide based medicine is on the horizon. Of those, RNAi or CRISPR/Cas- based therapies show great promise in combating various types of diseases caused by viral and parasitic agents that effect aquatic and fish medicine. In our modern times, when the marine industry has become so vital for feed and economic stability, even the most extreme alternative treatment strategies such as the use of small molecules or even the use of disease to control invasive species populations should be considered.
Michael Gotesman; Simon Menanteau-Ledouble; Mona Saleh; Sven M. Bergmann; Mansour El-Matbouli. A new age in AquaMedicine: unconventional approach in studying aquatic diseases. BMC Veterinary Research 2018, 14, 178 .
AMA StyleMichael Gotesman, Simon Menanteau-Ledouble, Mona Saleh, Sven M. Bergmann, Mansour El-Matbouli. A new age in AquaMedicine: unconventional approach in studying aquatic diseases. BMC Veterinary Research. 2018; 14 (1):178.
Chicago/Turabian StyleMichael Gotesman; Simon Menanteau-Ledouble; Mona Saleh; Sven M. Bergmann; Mansour El-Matbouli. 2018. "A new age in AquaMedicine: unconventional approach in studying aquatic diseases." BMC Veterinary Research 14, no. 1: 178.
Cyprinid herpesvirus 3 (CyHV-3) is the archetypal fish alloherpesvirus and the etiologic agent of a lethal disease in common and koi carp. To date, the genome sequences of only four CyHV-3 isolates have been published, but no comparisons of the biologic properties of these strains have been reported. We have sequenced the genomes of a further seven strains from various geographical sources, and have compared their growth in vitro and virulence in vivo. The major findings were: (i) the existence of the two genetic lineages previously described as European and Asian was confirmed, but inconsistencies between the geographic origin and genotype of some strains were revealed; (ii) potential inter-lineage recombination was detected in one strain, which also suggested the existence of a third, as yet unidentified lineage; (iii) analysis of genetic disruptions led to the identification of non-essential genes and their potential role in virulence; (iv) comparison of the in vitro and in vivo properties of strains belonging to the two lineages revealed that inter-lineage polymorphisms do not contribute to the differences in viral fitness observed; and (v) a negative correlation was observed among strains between viral growth in vitro and virulence in vivo. This study illustrates the importance of coupling genomic and biologic comparisons of viral strains in order to enhance understanding of viral evolution and pathogenesis.
Yuan Gao; Nicolás M. Suárez; Gavin S. Wilkie; Chuanfu Dong; Sven Bergmann; Pei-Yu Alison Lee; Andrew J. Davison; Alain F. C. Vanderplasschen; Maxime Boutier. Genomic and biologic comparisons of cyprinid herpesvirus 3 strains. Veterinary Research 2018, 49, 1 -11.
AMA StyleYuan Gao, Nicolás M. Suárez, Gavin S. Wilkie, Chuanfu Dong, Sven Bergmann, Pei-Yu Alison Lee, Andrew J. Davison, Alain F. C. Vanderplasschen, Maxime Boutier. Genomic and biologic comparisons of cyprinid herpesvirus 3 strains. Veterinary Research. 2018; 49 (1):1-11.
Chicago/Turabian StyleYuan Gao; Nicolás M. Suárez; Gavin S. Wilkie; Chuanfu Dong; Sven Bergmann; Pei-Yu Alison Lee; Andrew J. Davison; Alain F. C. Vanderplasschen; Maxime Boutier. 2018. "Genomic and biologic comparisons of cyprinid herpesvirus 3 strains." Veterinary Research 49, no. 1: 1-11.
A novel permanently growing brain cell line from koi (Cyprinus carpio L.) (KB cell line) was established, and its suitability for detection of koi herpesvirus (KHV) was demonstrated in this study. The KB cell line was optimally maintained at 27°C in Leibovitz's L‐15 medium supplemented with 10% foetal bovine serum (FBS). It was subcultured more than 100 times, and chromosome analysis revealed that 51.54% of KB cells at passage 80 maintained the abnormal diploid chromosome number 2n = 96 while the modal chromosome number was 2n = 100. The cell line was cryopreserved in liquid nitrogen at −196°C and was recovered from storage after 1 year with good cell viability and vitality. The results of virus isolation demonstrated that KB cells were susceptible to KHV, which was shown by the presence of an obvious cytopathic effect and abundant virus particles. The viral titres of KHV in KB reached 105.73TCID50/0.1 ml within 7 days. Immunofluorescence and Western blot assays confirmed that KB replicated KHV. The newly established KB cell line will serve as a useful tool to elucidate KHV disease (KHVD) pathogenesis.
Y Wang; W Zeng; Q Wang; Y Li; S M Bergmann; S Zheng; Y Ren; C Liu; O Chang; P Lee. Establishment and characterization of a new cell line from koi brain (Cyprinus carpio L.). Journal of Fish Diseases 2017, 41, 357 -364.
AMA StyleY Wang, W Zeng, Q Wang, Y Li, S M Bergmann, S Zheng, Y Ren, C Liu, O Chang, P Lee. Establishment and characterization of a new cell line from koi brain (Cyprinus carpio L.). Journal of Fish Diseases. 2017; 41 (2):357-364.
Chicago/Turabian StyleY Wang; W Zeng; Q Wang; Y Li; S M Bergmann; S Zheng; Y Ren; C Liu; O Chang; P Lee. 2017. "Establishment and characterization of a new cell line from koi brain (Cyprinus carpio L.)." Journal of Fish Diseases 41, no. 2: 357-364.
Worldwide koi herpesvirus (KHV) causes high mortalities in Cyprinus carpio L. aquaculture. So far, it is unknown how the different variants of KHV have developed and how they spread in the fish, but also in the environmental water bodies. Therefore, a phylogenetic method based on variable number of tandem repeats (VNTR) was improved to gain deeper insights into the phylogeny of KHV and its possible worldwide distribution. Moreover, a VNTR-3 qPCR was designed which allows fast virus typing. This study presents a useful method for molecular tracing of diverse KHV variants and lineages based on in vitro experiments. Including these techniques on can observe samples from natural sources.
Sandro Klafack; Qing Wang; Weiwei Zeng; Yingying Wang; Yingying Li; Shucheng Zheng; Jolanta Kempter; Pei-Yu Lee; Marek Matras; Sven M. Bergmann. Genetic Variability of Koi Herpesvirus In vitro—A Natural Event? Frontiers in Microbiology 2017, 8, 982 .
AMA StyleSandro Klafack, Qing Wang, Weiwei Zeng, Yingying Wang, Yingying Li, Shucheng Zheng, Jolanta Kempter, Pei-Yu Lee, Marek Matras, Sven M. Bergmann. Genetic Variability of Koi Herpesvirus In vitro—A Natural Event? Frontiers in Microbiology. 2017; 8 ():982.
Chicago/Turabian StyleSandro Klafack; Qing Wang; Weiwei Zeng; Yingying Wang; Yingying Li; Shucheng Zheng; Jolanta Kempter; Pei-Yu Lee; Marek Matras; Sven M. Bergmann. 2017. "Genetic Variability of Koi Herpesvirus In vitro—A Natural Event?" Frontiers in Microbiology 8, no. : 982.
Koi herpesvirus (KHV) causes KHV disease (KHVD). The virus is highly contagious in carp or koi and can induce a high mortality. Latency and, in some cases, a lack of signs presents a challenge for virus detection. Appropriate immunological detection methods for anti‐KHV antibodies have not yet been fully validated for KHV. Therefore, it was developed and validated an enzyme‐linked immunosorbent assay (ELISA) to detect KHV antibodies. The assay was optimized with respect to plates, buffers, antigens and assay conditions. It demonstrated high diagnostic and analytical sensitivity and specificity and was particularly useful at the pond or farm levels. Considering the scale of the carp and koi industry worldwide, this assay represents an important practical tool for the indirect detection of KHV, also in the absence of clinical signs.
S M Bergmann; Q Wang; W Zeng; Y Li; Y Wang; M Matras; M Reichert; D Fichtner; M Lenk; T Morin; N J Olesen; H F Skall; P-Y Lee; S Zheng; S Monaghan; S Reiche; W Fuchs; M Kotler; K Way; G Bräuer; K Böttcher; A Kappe; J Kielpinska. Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA). Journal of Fish Diseases 2017, 40, 1511 -1527.
AMA StyleS M Bergmann, Q Wang, W Zeng, Y Li, Y Wang, M Matras, M Reichert, D Fichtner, M Lenk, T Morin, N J Olesen, H F Skall, P-Y Lee, S Zheng, S Monaghan, S Reiche, W Fuchs, M Kotler, K Way, G Bräuer, K Böttcher, A Kappe, J Kielpinska. Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA). Journal of Fish Diseases. 2017; 40 (11):1511-1527.
Chicago/Turabian StyleS M Bergmann; Q Wang; W Zeng; Y Li; Y Wang; M Matras; M Reichert; D Fichtner; M Lenk; T Morin; N J Olesen; H F Skall; P-Y Lee; S Zheng; S Monaghan; S Reiche; W Fuchs; M Kotler; K Way; G Bräuer; K Böttcher; A Kappe; J Kielpinska. 2017. "Validation of a KHV antibody enzyme-linked immunosorbent assay (ELISA)." Journal of Fish Diseases 40, no. 11: 1511-1527.
Outbreaks of koi sleepy disease (KSD) caused by carp edema virus (CEV) may seriously affect populations of farmed common carp, one of the most important fish species for global food production. The present study shows further evidence for the involvement of CEV in outbreaks of KSD among carp and koi populations: in a series of infection experiments, CEV from two different genogroups could be transmitted to several strains of naïve common carp via cohabitation with fish infected with CEV. In recipient fish, clinical signs of KSD were induced. The virus load and viral gene expression results confirm gills as the target organ for CEV replication. Gill explants also allowed for a limited virus replication in vitro. The in vivo infection experiments revealed differences in the virulence of the two CEV genogroups which were associated with infections in koi or in common carp, with higher virulence towards the same fish variety as the donor fish. When the susceptibility of different carp strains to a CEV infection and the development of KSD were experimentally investigated, Amur wild carp showed to be relatively more resistant to the infection and did not develop clinical signs for KSD. However, the resistance could not be related to a higher magnitude of type I IFN responses of affected tissues. Despite not having a mechanistic explanation for the resistance of Amur wild carp to KSD, we recommend using this carp strain in breeding programs to limit potential losses caused by CEV in aquaculture.
Mikolaj Adamek; Anna Oschilewski; Peter Wohlsein; Verena Jung-Schroers; Felix Teitge; Andy Dawson; David Gela; Veronika Piackova; Martin Kocour; Jerzy Adamek; Sven M. Bergmann; Dieter Steinhagen. Experimental infections of different carp strains with the carp edema virus (CEV) give insights into the infection biology of the virus and indicate possible solutions to problems caused by koi sleepy disease (KSD) in carp aquaculture. Veterinary Research 2017, 48, 1 -16.
AMA StyleMikolaj Adamek, Anna Oschilewski, Peter Wohlsein, Verena Jung-Schroers, Felix Teitge, Andy Dawson, David Gela, Veronika Piackova, Martin Kocour, Jerzy Adamek, Sven M. Bergmann, Dieter Steinhagen. Experimental infections of different carp strains with the carp edema virus (CEV) give insights into the infection biology of the virus and indicate possible solutions to problems caused by koi sleepy disease (KSD) in carp aquaculture. Veterinary Research. 2017; 48 (1):1-16.
Chicago/Turabian StyleMikolaj Adamek; Anna Oschilewski; Peter Wohlsein; Verena Jung-Schroers; Felix Teitge; Andy Dawson; David Gela; Veronika Piackova; Martin Kocour; Jerzy Adamek; Sven M. Bergmann; Dieter Steinhagen. 2017. "Experimental infections of different carp strains with the carp edema virus (CEV) give insights into the infection biology of the virus and indicate possible solutions to problems caused by koi sleepy disease (KSD) in carp aquaculture." Veterinary Research 48, no. 1: 1-16.
Hygienic measures such as disinfection are important tools for the maintenance of fish health in aquaculture. While little information is available on the disinfection of water intended for fish containment, Huwa-San®, a disinfectant used in food and water industries, was used for daily treatment at concentrations of approximately 60 ppm over a total period of 3 months (experiment 1) with a 3-week treatment-free interval after 2 months (experiment 2). During this period, koi herpesvirus (KHV) was added to the water of two aquaria, one used as a normal contact control, the other one receiving daily water disinfectant treatments that prevented KHV infection of carp. In the second experiment, Huwa-San® treatment was interrupted and KHV infection was prevalent. However, when naïve fish were introduced to the same aquarium after re-application of disinfectant, KHV could not be detected in those naïve fish. Whilst KHV could not be detected in samples where disinfectant had been applied, it was present in samples of naïve fish cohabiting with infection contact control animals which had undergone no disinfectant treatment over experiments 1 and 2. The results presented here show that water treatment with a disinfectant may prevent transmission of infectious KHV to naïve carp cohabited with infected carp.
S M Bergmann; E S Monro; J Kempter. Can water disinfection prevent the transmission of infectious koi herpesvirus to naïve carp? - a case report. Journal of Fish Diseases 2016, 40, 885 -893.
AMA StyleS M Bergmann, E S Monro, J Kempter. Can water disinfection prevent the transmission of infectious koi herpesvirus to naïve carp? - a case report. Journal of Fish Diseases. 2016; 40 (7):885-893.
Chicago/Turabian StyleS M Bergmann; E S Monro; J Kempter. 2016. "Can water disinfection prevent the transmission of infectious koi herpesvirus to naïve carp? - a case report." Journal of Fish Diseases 40, no. 7: 885-893.
The KHV free carp were experimentally infected with CyHV-3 suspension by immersion at temperatures of 18ºC±1 and 25ºC±1. Skin, brain, gills, liver, spleen, kidneys, and leukocytes were investigated for the presence of KHV DNA, and sera for the presence of antibodies against the virus. Similar investigations were performed with non-infected carp, designed as control groups. The results of KHV DNA detection in the infected carp kept at 25ºC ±1 were compared with the results of the virus isolation onto common carp brain cells using leukocyte co-cultivation method and results of specific antibody detection. The appearance of specific antibodies at various time intervals after detecting the clinical symptoms of KHVD in naturally infected carp was additionally studied. At 18°C water temperature, KHV DNA was not detected up to the 6th d post infection (dpi). Then it was found in samples from skin, gill, liver, spleen, and kidneys and much later from the brain. At 25°C water temperature, KHV DNA was already detected on the 3rd dpi in samples from skin and gills, and from the 5th dpi it was found in all examined internal organs, except the brain where it was found at 6th dpi. At 25°C, isolations of CyHV-3 have succeeded between the 7th and 11th dpi. The first specific antibodies were found no sooner than on the 21st dpi. The serological examination performed in naturally infected carp in 15 carp pond farms showed that first specific antiviral antibodies can be detected 2 weeks after the appearance of clinical symptoms of KHVD.
Marek Matras; Jerzy Antychowicz; Jeannette Castric; Sven M. Bergmann. CYHV-3 Infection Dynamics in Common Carp (Cyprinus Carpio) – Evaluation of Diagnostic Methods. Bulletin of the Veterinary Institute in Pulawy 2012, 56, 127 -132.
AMA StyleMarek Matras, Jerzy Antychowicz, Jeannette Castric, Sven M. Bergmann. CYHV-3 Infection Dynamics in Common Carp (Cyprinus Carpio) – Evaluation of Diagnostic Methods. Bulletin of the Veterinary Institute in Pulawy. 2012; 56 (2):127-132.
Chicago/Turabian StyleMarek Matras; Jerzy Antychowicz; Jeannette Castric; Sven M. Bergmann. 2012. "CYHV-3 Infection Dynamics in Common Carp (Cyprinus Carpio) – Evaluation of Diagnostic Methods." Bulletin of the Veterinary Institute in Pulawy 56, no. 2: 127-132.