A mézelő méh (Apis mellifera L.) vírusfertőzéseinek és a méhpatogén vírusok molekuláris szerkezetének tanulmányozása = Investigations on the viral infections of the honey bee (Apis mellifera L.) and on the molecular structure of bee pathogens

A mézelő méh vírusainak vizsgálatára irányuló kutatások keretében polimeráz láncreakcióra (PCR) alapozott felmérő vizsgálatokat végeztünk magyarországi és ausztriai méhészetekben a hat legfontosabb vírus előfordulási gyakoriságának megismerésére. Elsőként mutattuk ki a méhek szárnydeformitását okozó vírus, a fekete anyabölcső vírus, a lárvatömlősödés vírus és a krónikus méhbénulás vírus előfordulását hazai és ausztriai méhészetekben. Eltéréseket tapasztaltunk a vírusok hazai és külföldi előfordulási gyakorisága között. Filogenetikai vizsgálatokat végeztünk a fekete anyabölcső vírus közép-európai törzseinek bevonásával és feltártuk a vírus különböző genomterületeinek változékonyságát. A méhek szárnydeformitását okozó vírus világszerte gyűjtött törzseinek filogenetikai összehasonlítása során nagyon közeli rokonságot tapasztaltunk a vizsgált szekvenciák között, amely a vírus közelmúltbeli gyors terjedésére utal. Ez a terjedés valószínűleg összefüggésben áll a Varroa destructor atka elterjedésével, mivel ez a parazita hatékony vektora a vírusnak. Meghatároztuk a heveny méhbénulás vírus egy magyarországi és egy lengyelországi törzsének teljes genomszekvenciáját. A zab levéltetű (Rhopalosiphum padi) vírusát mutattuk ki méh eredetű mintákból. Folytattuk a hazai méhekből korábban kimutatott, a heveny méhbénuláshoz és a Kashmir méhvírushoz hasonló vírus genetikai jellemzését. Három méhvírus egyidejű kimutatására alkalmas multiplex RT-PCR alapú diagnosztikai módszert fejlesztettünk ki. | The project was focused on the investigations of honeybee viruses. Polymerase chain reaction (PCR)-based surveillance was performed on bee samples from Hungarian and Austrian apiaries to record the incidence of the six most important bee viruses. We have detected the deformed wing virus (DWV), the black queen cell virus (BQCV), the sacbrood virus, and the chronic bee paralysis virus for the first time in these countries. We found considerable differences in the prevalence of these viruses in different countries. Phylogenetic analyses were made on central European BQCV strains, and the variability of the different genome regions was recorded. By the genetic analysis of DWV strains collected worldwide, we detected close genetic relationship between the strains, which indicates a recent spread of the virus. This is probably in connection with the global spread of Varroa destructor, because this parasitic mite is an efficient vector of the virus. We have determined the complete genome sequence of a Hungarian and a Polish acute bee paralysis virus (ABPV) strain. We have detected the bird cherry aphid (Rhopalosiphum padi) virus in bee samples. We carried on the genetic characterization of a bee virus, which was detected in Hungary, and which is closely related to the ABPV and the Kashmir bee virus. We have developed a multiplex RT-PCR assay for the simultaneous detection of three viruses in honeybee samples

Analysis of a novel RNA virus in a wild northern white-breasted hedgehog (Erinaceus roumanicus)

Tombusviruses are generally considered plant viruses. A novel tombus-/carmotetravirus-like RNA virus was identified in a faecal sample and blood and muscle tissues from a wild northern white-breasted hedgehog (Erinaceus roumanicus). The complete genome of the virus, called H14-hedgehog/2015/HUN (GenBank accession number MN044446), is 4,118 nucleotides in length with a readthrough stop codon of type/group 1 in ORF1 and lacks a poly(A) tract at the 3' end. The predicted ORF1-RT (RdRp) and the capsid proteins had low (31-33%) amino acid sequence identity to unclassified tombus-/noda-like viruses (Hubei tombus-like virus 12 and Beihai noda-like virus 10), respectively, discovered recently in invertebrate animals. An in vivo experimental plant inoculation study showed that an in vitro-transcribed H14-hedgehog/2015/HUN viral RNA did not replicate in Nicotiana benthamiana, Chenopodium quinoa, or Chenopodium murale, the most susceptible hosts for plant-origin tombusviruses

Eco-epidemiology of Borrelia miyamotoi and Lyme borreliosis spirochetes in a popular hunting and recreational forest area in Hungary

BACKGROUND: Borrelia miyamotoi, the newly discovered human pathogenic relapsing fever spirochete, and Borrelia burgdorferi sensu lato are maintained in natural rodent populations. The aim of this study was to investigate the natural cycle of B. miyamotoi and B. burgdorferi s.l. in a forest habitat with intensive hunting, forestry work and recreational activity in Southern Hungary. METHODS: We collected rodents with modified Sherman-traps during 2010–2013 and questing ticks with flagging in 2012. Small mammals were euthanized, tissue samples were collected and all ectoparasites were removed and stored. Samples were screened for pathogens with multiplex quantitative real-time polymerase chain reaction (qPCR) targeting a part of flagellin gene, then analysed with conventional PCRs and sequencing. RESULTS: 177 spleen and 348 skin samples of six rodent species were individually analysed. Prevalence in rodent tissue samples was 0.2 % (skin) and 0.5 % (spleen) for B. miyamotoi and 6.6 % (skin) and 2.2 % (spleen) for B. burgdorferi s.l. Relapsing fever spirochetes were detected in Apodemus flavicollis males, B. burgdorferi s.l. in Apodemus spp. and Myodes glareolus samples. Borrelia miyamotoi was detected in one questing Ixodes ricinus nymph and B. burgdorferi s.l in nymphs and adults. In the ticks removed from rodents DNA amplification of both pathogens was successful from I. ricinus larvae (B. miyamotoi 5.6 %, B. burgdorferi s.l. 11.1 %) and one out of five nymphs while from Ixodes acuminatus larvae, and nymph only B. burgdorferi s.l. DNA was amplified. Sequencing revealed B. lusitaniae in a questing I. ricinus nymph and altogether 17 B. afzelii were identified in other samples. Two Dermacentor marginatus engorged larva pools originating from uninfected hosts were also infected with B. afzelii. CONCLUSIONS: This is the first report of B. miyamotoi occurrence in a natural population of A. flavicollis as well as in Hungary. We provide new data about circulation of B. burgdorferi s.l. in rodent and tick communities including the role of I. acuminatus ticks in the endophilic pathogen cycle. Our results highlight the possible risk of infection with relapsing fever and Lyme borreliosis spirochetes in forest habitats especially in the high-risk groups of hunters, forestry workers and hikers

Tick paralysis induced by Ixodes gibbosus: enigmatic cases in domestic mammals from Cyprus

Tick paralysis is a potentially fatal condition caused by toxins produced and secreted by tick salivary glands. This survey presents clinical and epidemiological observations of tick paralysis cases in domestic animals in Cyprus. Local veterinarians report typical tick paralysis cases occurring in goats, sheep, dogs, and cats. The animals suffering from paralysis are free from other neurological diseases, have blood and biochemical parameters within normal ranges, and recover fast by simply removing the ticks found predominantly on the head and around the neck. Tick paralysis cases occur in a specific geographic area of Cyprus (Akamas peninsula), from September through March, but not every year. Instead, the phenomenon has 2 periodic cycles of occurrence, a 3- and a 7-year cycle. The 2 cycles are differentiated by severity based on the number of affected animals and the resulting losses. As described for other tick-borne diseases, these cyclic patterns may be attributed to external factors, self-oscillations of the disease system, or the combined action of these mechanisms. Ticks collected from a recent paralysis case in a goat were morphologically and molecularly identified as Ixodes gibbosus. Efforts should be made to characterize the specific toxins involved in tick paralysis and to develop a vaccine, which could prevent significant losses of small ruminants, especially in free-ranging farming systems, a prevalent management approach observed in Cyprus and various regions worldwide

The 3P Framework: A Comprehensive Approach to Coping with the Emerging Infectious Disease Crisis

The COVID-19 pandemic is the latest example of the profound socioeconomic impact of the emerging infectious disease (EID) crisis. Current health security measures are based on a failed evolutionary paradigm that presumes EID is rare and cannot be predicted because emergence requires the prior evolution of novel genetic capacities for colonizing a new host. Consequently, crisis response through preparation for previously emerged diseases and palliation following outbreaks have been the only health security options, which have become unsustainably expensive and unsuccessful. The Stockholm paradigm (SP) is an alternative evolutionary framework that suggests host changes are the result of changing conditions that bring pathogens into contact with susceptible hosts, with novel genetic variants arising in the new host after infection. Host changes leading to EID can be predicted because preexisting capacities for colonizing new hosts are highly specific and phylogenetically conservative. This makes EID prevention through limiting exposure to susceptible hosts possible. The DAMA (Document, Assess, Monitor, Act) protocol is a policy extension of the SP that can both prevent and mitigate EID by enhancing traditional efforts through adding early warning signs and predicting transmission dynamics. Prevention, preparation, and palliation compose the 3P framework, a comprehensive plan for reducing the socioeconomic impact of EID. This article has been produced in support of and with appreciation for the efforts by Gábor Földvári of the Institute of Evolution, Centre for Ecological Research, and the Centre for Eco-Epidemiology, National Laboratory for Health Security (both located at 1121 Budapest, Konkoly-Thege Miklós út 29-33, Hungary). Through his untiring efforts, team building, and leadership, he has secured the first EU-wide team research grant. This work was supported by the National Research, Development and Innovation Office in Hungary (RRF-2.3.1-21-2022-00006) and the COST Action CA21170 “Prevention, anticipation and mitigation of tick-borne disease risk applying the DAMA protocol (PRAGMATICK),” which represent the first funded efforts to apply the principles of the DAMA protocol