A novel adenovirus of Western lowland gorillas (Gorilla gorilla gorilla)

Adenoviruses (AdV) broadly infect vertebrate hosts including a variety of primates. We identified a novel AdV in the feces of captive gorillas by isolation in cell culture, electron microscopy and PCR. From the supernatants of infected cultures we amplified DNA polymerase (DPOL), preterminal protein (pTP) and hexon gene sequences with generic pan primate AdV PCR assays. The sequences in-between were amplified by long-distance PCRs of 2 - 10 kb length, resulting in a final sequence of 15.6 kb. Phylogenetic analysis placed the novel gorilla AdV into a cluster of primate AdVs belonging to the species Human adenovirus B (HAdV-B). Depending on the analyzed gene, its position within the cluster was variable. To further elucidate its origin, feces samples of wild gorillas were analyzed. AdV hexon sequences were detected which are indicative for three distinct and novel gorilla HAdV-B viruses, among them a virus nearly identical to the novel AdV isolated from captive gorillas. This shows that the discovered virus is a member of a group of HAdV-B viruses that naturally infect gorillas. The mixed phylogenetic clusters of gorilla, chimpanzee, bonobo and human AdVs within the HAdV-B species indicate that host switches may have been a component of the evolution of human and non-human primate HAdV-B viruses

Novel polyomaviruses in human and non-human primates

Polyomaviren sind kleine, unbehüllte DNA-Viren, die sowohl Säugetiere als auch Vögel infizieren. Sie sind in der Lage, schwere Erkrankungen bei immungeschwächten Personen zu verursachen. Angesichts der zunehmenden Anzahl immunsupprimierter Menschen gelangen Polyomaviren als aufstrebende opportunistische Erreger zunehmend in den Fokus wissenschaftlicher Forschung. Um Einblicke in die Vielfalt und Verbreitung dieser Viren zu gewinnen, behandelt die vorliegende Arbeit folgende Fragestellungen: (i) Existieren weitere Polyomaviren in humanen und nichthumanen Primaten? (ii) Wie gliedern sich diese Vertreter in den phylogenetischen Stammbaum der Polyomaviren ein? (iii) Liefern diese Viren Hinweise auf mögliche Transmissionswege, Tropismen und Pathogenitäten der Polyomaviren? (iv) Inwiefern lassen Polyomaviren in nicht-humanen Primaten auf die Existenz bisher unbekannter menschlicher Polyomaviren schließen? Es wurden 792 Nekropsieproben simianer Herkunft mittels degenerierter PCR analysiert und auf Polyomavirus-Sequenzen untersucht. Milz-, Lymphknoten- und Darmproben erwiesen sich als geeignetes Material zum Polyomavirus-Nachweis. Es konnten 30 neuartige nicht-humane Primaten-Polyomaviren identifiziert werden: 19 in Menschenaffen (15 in Schimpansen, drei in Gorillas und eins im Orang-Utan), fünf in Altweltaffen und sechs in Neuweltaffen. Siebzehn Komplettgenome wurden erfolgreich sequenziert. Die phylogenetische Analyse zeigt, dass diese Polyomaviren über den gesamten Stammbaum verteilt sind. Zehn Polyomaviren aus wilden Menschenaffen weisen eine nahe Verwandtschaft zum humanen MCPyV auf. Dies lässt vermuten, dass MCPyV aus der Übertragung eines MCPyV-ähnlichen Schimpansen Polyomavirus auf den Menschen hervorgegangen ist. Zusätzlich wurden 597 humane Proben mit Hilfe einer degenerierten PCR untersucht, wobei sich Urin als Probenmaterial zum Polyomavirus-Nachweis gut eignete. Im Serum einer nierentransplantierten Patientin, die sich unter immunsuppressiver Behandlung befand, konnte ein bisher unbekanntes humanes Polyomavirus detektiert und das Genom vollständig sequenziert werden. Da bereits acht humane Polyomaviren bekannt waren, wurde dieses Virus Humanes Polyomavirus 9 (HPyV9) genannt. Im phylogenetischen Stammbaum ist es nahe mit dem LPyV verwandt. Humane Seroreaktivitäten gegen LPyV wurden auf eine Kreuzreaktivität zwischen HPyV9 und LPyV zurückgeführt. Um die Vielfalt der menschlichen Polyomaviren weiter zu untersuchen, wurde ein kombinierter Ansatz verwendet. Neben der Identifizierung und phylogenetischen Analyse der Affen-Polyomaviren wurden zusätzlich humane Seren auf Antikörper gegen Schimpansen-Polyomaviren untersucht. Das VP1 von vier Schimpansen-Polyomaviren, die phylogenetisch keinen humanen polyomaviralen Verwandten besitzen, wurde in E. coli exprimiert und als Antigen in einem ELISA verwendet. Humane Serum- und Plasmaproben aus der Elfenbeinküste und aus Deutschland zeigten zum Teil starke Seroreaktivitäten gegen die vier VP1 der Schimpansen-Polyomaviren. Es wird daher postuliert, dass innerhalb der menschlichen Population weitere Polyomaviren existieren, die genetisch mit den untersuchten Schimpansen- Polyomaviren verwandt sind und serologisch kreuzreagieren. Die Ergebnisse dieser Arbeit leisten einen wichtigen Beitrag, sowohl zum Verständnis der Diversität als auch zum Tropismus und zur evolutionären Entwicklung der Polyomaviren. Die generierten Sequenzinformationen ermöglichen die Entwicklung effektiverer nukleinsäure-basierter Systeme zur Detektion von bisher unbekannten Polyomaviren.Polyomaviruses are a family of small non-enveloped DNA viruses. They infect a wide range of birds and mammals and are able to cause severe diseases in immunocompromised individuals. Given the growing disease burden entailed by acquired immunodeficiencies, human PyVs are now increasingly considered emerging opportunistic pathogens. Gaining more insight into their diversity, prevalence, and etiopathogenesis is therefore essential. The present study assessed the following questions: (i) Are additionalpolyomaviruses circulating in humans and non-human primates? (ii) How is their phylogenetic relationship to other polyomaviruses? (iii) Are these viruses able to provide insight into possible transmission routes, tropisms and pathogenities? (iv) Is there evidence for the existence of so far unknown human polyomaviruses? A total of 792 necropsy samples of simian origin were analyzed for the presence of polyomaviruses by using degenerate primer-based PCR. With this approach a high prevalence of polyomaviruses in spleen, lymph node and intestine samples was detected and 30 novel non-human primate polyomaviruses were identified: 19 in great apes (15 in chimpanzees, three in gorillas and one in orangutan), five in Old World monkeys and six in New World monkeys. Seventeen complete genomes were amplified. Phylogenetic analysis revealed that these new polyomaviruses span nearly the entire known diversity of mammalian polyomaviruses. This fact suggests that primates as a whole, including humans, are infected with a plethora of polyomaviruses. Ten polyomaviruses detected in wild Great apes revealed a remarkably close relationship to the human MCPyV. Thus, MCPyV could be the result of interspecies transmission of a MCPyV-like chimpanzee polyomavirus to humans. Additional 597 human samples were tested by using a degenerate PCR, wherein urine samples proved as the most appropriate material for polyomavirus detection. An unknown polyomavirus sequence was amplified from the serum of a kidney transplant patient under immunosuppressive treatment. The genome of this virus was completely sequenced. In phylogenetic analyses, it appeared as the closest relative to the African green monkey- derived lymphotropic polyomavirus (LPyV). The reactivity of human sera against LPyV is due to crossreactivity between HPyV9 and LPyV. To further examine the diversity of human polyomaviruses we used a combinatorial approach comprised of initial degenerate primer-based PCR identification and phylogenetic analysis of non-human primate species. In addition, polyomavirus specific serological analysis of human sera was applied. Four chimpanzee polyomaviruses with no human counterpart were expressed in E. coli for use as antigens in an ELISA. Human serum and plasma samples from both Côte d‘Ivoire and Germany showed frequent seropositivity for these four viruses. These results support the existence of additional polyomaviruses circulating within the human population that are genetically and serologically related to existing chimpanzee polyomaviruses. Given the accelerated rate of human polyomavirus discovery over the last few years, it appears very likely that further, still- unknown polyomaviruses are actually circulating in human populations. Elucidating the evolutionary development may foster a better understanding of the pathogenicity of the novel polyomaviruses. Therefore, our study on primates is of importance since it provides insight into polyomavirus diversity and tropism due to improved nucleic-based methods for the detection of unknown polyomaviruses

Whole-Genome Investigation of <i>Salmonella</i> Dublin Considering Mountain Pastures as Reservoirs in Southern Bavaria, Germany

Worldwide, Salmonella Dublin (S. Dublin) is responsible for clinical disease in cattle and also in humans. In Southern Bavaria, Germany, the serovar was identified as a causative agent for 54 animal disease outbreaks in herds between 2017 and 2021. Most of these emerged from cattle herds (n = 50). Two occurred in pig farms and two in bovine herds other than cattle. Genomic analysis of 88 S. Dublin strains isolated during these animal disease outbreaks revealed 7 clusters with 3 different MLST-based sequence types and 16 subordinate cgMLST-based complex types. Antimicrobial susceptibility investigation revealed one resistant and three intermediate strains. Furthermore, only a few genes coding for bacterial virulence were found among the isolates. Genome analysis enables pathogen identification and antimicrobial susceptibility, serotyping, phylogeny, and follow-up traceback analysis. Mountain pastures turned out to be the most likely locations for transmission between cattle of different herd origins, as indicated by epidemiological data and genomic traceback analyses. In this context, S. Dublin shedding was also detected in asymptomatic herding dogs. Due to the high prevalence of S. Dublin in Upper Bavaria over the years, we suggest referring to this administrative region as “endemic”. Consequently, cattle should be screened for salmonellosis before and after mountain pasturing

A novel adenovirus of Western lowland gorillas (<it>Gorilla gorilla gorilla</it>)

Abstract Adenoviruses (AdV) broadly infect vertebrate hosts including a variety of primates. We identified a novel AdV in the feces of captive gorillas by isolation in cell culture, electron microscopy and PCR. From the supernatants of infected cultures we amplified DNA polymerase (DPOL), preterminal protein (pTP) and hexon gene sequences with generic pan primate AdV PCR assays. The sequences in-between were amplified by long-distance PCRs of 2 - 10 kb length, resulting in a final sequence of 15.6 kb. Phylogenetic analysis placed the novel gorilla AdV into a cluster of primate AdVs belonging to the species Human adenovirus B (HAdV-B). Depending on the analyzed gene, its position within the cluster was variable. To further elucidate its origin, feces samples of wild gorillas were analyzed. AdV hexon sequences were detected which are indicative for three distinct and novel gorilla HAdV-B viruses, among them a virus nearly identical to the novel AdV isolated from captive gorillas. This shows that the discovered virus is a member of a group of HAdV-B viruses that naturally infect gorillas. The mixed phylogenetic clusters of gorilla, chimpanzee, bonobo and human AdVs within the HAdV-B species indicate that host switches may have been a component of the evolution of human and non-human primate HAdV-B viruses.</p

African Great Apes Are Naturally Infected with Polyomaviruses Closely Related to Merkel Cell Polyomavirus

The oncogenic Merkel cell polyomavirus (MCPyV) infects humans worldwide, but little is known about the occurrence of viruses related to MCPyV in the closest phylogenetic relatives of humans, great apes. We analyzed samples from 30 wild chimpanzees and one captive gorilla and identified two new groups of polyomaviruses (PyVs). These new viruses are by far the closest relatives to MCPyV described to date, providing the first evidence of the natural occurrence of PyVs related to MCPyV in wild great apes. Similar to MCPyV, the prevalence of these viruses is relatively high (>30%). This, together with the fact that humans in West and Central Africa frequently hunt and butcher primates, may point toward further MCPyV-like strains spreading to, or already existing in, our species

White-Toothed Shrews (Genus <i>Crocidura</i>): Potential Reservoirs for Zoonotic <i>Leptospira</i> spp. and Arthropod-Borne Pathogens?

Three species of white-toothed shrews of the order Eulipotyphla are present in central Europe: the bicolored (Crocidura leucodon), greater (Crocidura russula) and lesser (Crocidura suaveolens) white-toothed shrews. Their precise distribution in Germany is ill-defined and little is known about them as reservoirs for zoonotic pathogens (Leptospira spp., Coxiella burnetii, Brucella spp., Anaplasma phagocytophilum, Babesia spp., Neoehrlichia mikurensis and Bartonella spp.). We investigated 372 Crocidura spp. from Germany (n = 341), Austria (n = 18), Luxembourg (n = 2) and Slovakia (n = 11). West European hedgehogs (Erinaceus europaeus) were added to compare the presence of pathogens in co-occurring insectivores. Crocidura russula were distributed mainly in western and C. suaveolens mainly in north-eastern Germany. Crocidura leucodon occurred in overlapping ranges with the other shrews. Leptospira spp. DNA was detected in 28/227 C. russula and 2/78 C. leucodon samples. Further characterization revealed that Leptospira kirschneri had a sequence type (ST) 100. Neoehrlichia mikurensis DNA was detected in spleen tissue from 2/213 C. russula samples. Hedgehogs carried DNA from L. kirschneri (ST 100), L. interrogans (ST 24), A. phagocytophilum and two Bartonella species. This study improves the knowledge of the current distribution of Crocidura shrews and identifies C. russula as carrier of Leptospira kirschneri. However, shrews seem to play little-to-no role in the circulation of the arthropod-borne pathogens investigated

Competitive inhibition of seroreactivity between human and chimpanzee polyomaviruses.

a<p>Before ELISA, 2 µg/ml of VP1 antigen was used for pre-adsorption of antibodies from human sera.</p>b<p>not tested.</p