RESUMO
The specificity of phages and their ability to evolve and overcome bacterial resistance make them potentially useful as adjuncts in the treatment of antibiotic-resistant bacterial infections. The goal of this study was to mimic a natural grouping of phages of interest and to evaluate the nature of their proliferation dynamics with bacteria. We have, for the first time, transferred naturally occurring phage groups directly from their sources of isolation to in vitro and identified 13 P. aeruginosa and 11 K. pneumoniae phages of 18 different genera, whose host range was grouped as 1.2-17%, 28-48% and 60-87%, using a large collection of P. aeruginosa (n = 102) and K. pneumoniae (n = 155) strains carrying different virulence factors and phage binding receptors. We introduced the interpretation model curve for phage liquid culturing, which allows easy and quick analysis of bacterial and phage co-proliferation and growth of phage-resistant mutants (PRM) based on qualitative and partially quantitative evaluations. We assayed phage lytic activities both individually and in 14 different cocktails on planktonic bacterial cultures, including three resistotypes of P. aeruginosa (PAO1, PA14 and PA7) and seven K. pneumoniae strains of different capsular serotypes. Based on the results, the natural phage cocktails designed and tested in this study largely performed well and inhibited PRM growth either synergistically or in proto-cooperation. This study contributes to the knowledge of phage behavior in cocktails and the formulation of therapeutic phage preparations. The paper also provides a detailed description of the methods of working with phages.
RESUMO
In contrast to the many reports of successful real-world cases of personalized bacteriophage therapy (BT), randomized controlled trials of non-personalized bacteriophage products have not produced the expected results. Here we present the outcomes of a retrospective observational analysis of the first 100 consecutive cases of personalized BT of difficult-to-treat infections facilitated by a Belgian consortium in 35 hospitals, 29 cities and 12 countries during the period from 1 January 2008 to 30 April 2022. We assessed how often personalized BT produced a positive clinical outcome (general efficacy) and performed a regression analysis to identify functional relationships. The most common indications were lower respiratory tract, skin and soft tissue, and bone infections, and involved combinations of 26 bacteriophages and 6 defined bacteriophage cocktails, individually selected and sometimes pre-adapted to target the causative bacterial pathogens. Clinical improvement and eradication of the targeted bacteria were reported for 77.2% and 61.3% of infections, respectively. In our dataset of 100 cases, eradication was 70% less probable when no concomitant antibiotics were used (odds ratio = 0.3; 95% confidence interval = 0.127-0.749). In vivo selection of bacteriophage resistance and in vitro bacteriophage-antibiotic synergy were documented in 43.8% (7/16 patients) and 90% (9/10) of evaluated patients, respectively. We observed a combination of antibiotic re-sensitization and reduced virulence in bacteriophage-resistant bacterial isolates that emerged during BT. Bacteriophage immune neutralization was observed in 38.5% (5/13) of screened patients. Fifteen adverse events were reported, including seven non-serious adverse drug reactions suspected to be linked to BT. While our analysis is limited by the uncontrolled nature of these data, it indicates that BT can be effective in combination with antibiotics and can inform the design of future controlled clinical trials. BT100 study, ClinicalTrials.gov registration: NCT05498363 .
Assuntos
Antibacterianos , Infecções Bacterianas , Bacteriófagos , Terapia por Fagos , Humanos , Estudos Retrospectivos , Terapia por Fagos/métodos , Bacteriófagos/fisiologia , Bacteriófagos/genética , Feminino , Masculino , Pessoa de Meia-Idade , Antibacterianos/uso terapêutico , Adulto , Infecções Bacterianas/terapia , Resultado do Tratamento , Idoso , Medicina de Precisão/métodos , Adolescente , Adulto Jovem , Bactérias/virologia , Bactérias/genética , Criança , Idoso de 80 Anos ou mais , Pré-Escolar , Bélgica , LactenteRESUMO
Phage therapy is recognized to be a promising alternative to fight antibiotic-resistant infections. In the quest for oral dosage forms containing bacteriophages, the utilization of colonic-release Eudragit® derivatives has shown potential in shielding bacteriophages from the challenges encountered within the gastrointestinal tract, such as fluctuating pH levels and the presence of digestive enzymes. Consequently, this study aimed to develop targeted oral delivery systems for bacteriophages, specifically focusing on colon delivery and employing Eudragit® FS30D as the excipient. The bacteriophage model used was LUZ19. An optimized formulation was established to not only preserve the activity of LUZ19 during the manufacturing process but also ensure its protection from highly acidic conditions. Flowability assessments were conducted for both capsule filling and tableting processes. Furthermore, the viability of the bacteriophages remained unaffected by the tableting process. Additionally, the release of LUZ19 from the developed system was evaluated using the Simulator of the Human Intestinal Microbial Ecosystem (SHIME®) model. Finally, stability studies demonstrated that the powder remained stable for at least 6 months when stored at +5 °C.
RESUMO
Hantaviruses (genus Hantavirus, family Bunyaviridae) cause hemorrhagic fever with renal syndrome (HFRS) in Eurasia and hantavirus (cardio)pulmonary syndrome (HCPS) in the Americas. So far, in Europe, four pathogenic hantaviruses have been found, often in co-circulation: Puumala virus (PUUV), Dobrava virus (DOBV), Saaremaa virus (SAAV), and Seoul virus (SEOV). Of those, only PUUV was found in Belgium. Recently, in our search for hantaviruses in the Flanders region of Belgium we collected genetic and serological evidence for the presence of SEOV virus in local brown rats. In this article, the results of (phylo)genetic analysis of wild-type SEOV strain from the Flanders are presented. The analysis based on the complete S segment sequence and also partial M- and L-segment sequences revealed that the Belgian SEOV strain was related most closely to strains from France, Indonesia, Japan, Singapore, Cambodia (those associated with the species Rattus norvegicus) and Vietnam. Such a clustering was in perfect agreement with the results of direct sequence comparison and suggested the same evolutionary history for all three genome segments of the Belgian SEOV strain (i.e., no reassortment of genome segments). So far, SEOV has been found in two European countries, France and Belgium, and there is every reason to believe that the area of the virus distribution in Europe is not restricted to those countries.
Assuntos
Febre Hemorrágica com Síndrome Renal/veterinária , Doenças dos Roedores/epidemiologia , Vírus Seoul/genética , Animais , Bélgica/epidemiologia , DNA Viral/análise , Febre Hemorrágica com Síndrome Renal/epidemiologia , Febre Hemorrágica com Síndrome Renal/virologia , Dados de Sequência Molecular , Filogenia , Ratos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Doenças dos Roedores/virologia , Vírus Seoul/classificação , Vírus Seoul/isolamento & purificação , Análise de Sequência de DNARESUMO
In the present study, we evaluated the effect of spray-drying formulations and operating parameters of a laboratory-scale spray-dryer on the characteristics of spray-dried powders containing two Pseudomonas aeruginosa bacteriophages exhibiting different morphotypes: a podovirus (LUZ19) and a myovirus (14-1). We optimized the production process for bacteriophage-loaded powders, with an emphasis on long-term storage under ICH (international conference on harmonization) conditions. D-trehalose-/L-isoleucine-containing bacteriophage mixtures were spray-dried from aqueous solutions using a Büchi Mini Spray-dryer B-290 (Flawil, Switzerland). A response surface methodology was used for the optimization of the spray-drying process, with the following as-evaluated parameters: Inlet temperature, spray gas flow rate, and the D-trehalose/L-isoleucine ratio. The dried powders were characterized in terms of yield, residual moisture content, and bacteriophage lytic activity. L-isoleucine has demonstrated a positive impact on the activity of LUZ19, but a negative impact on 14-1. We observed a negligible impact of the inlet temperature and a positive correlation of the spray gas flow rate with bacteriophage activity. After optimization, we were able to obtain dry powder preparations of both bacteriophages, which were stable for a minimum of one year under different ICH storage conditions (up to and including 40 °C and 75% relative humidity).
Assuntos
Bacteriófagos , Dessecação/métodos , Myoviridae , Podoviridae , Pseudomonas aeruginosa/virologia , Bacteriófagos/fisiologia , Liofilização/métodos , Myoviridae/fisiologia , Podoviridae/fisiologia , Pós , Suíça , TemperaturaRESUMO
More than a year after the first identification of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) as the causative agent of the 2019 coronavirus disease (COVID-19) in China, the emergence and spread of genomic variants of this virus through travel raise concerns regarding the introduction of lineages in previously unaffected regions, requiring adequate containment strategies. Concomitantly, such introductions fuel worries about a possible increase in transmissibility and disease severity, as well as a possible decrease in vaccine efficacy. Military personnel are frequently deployed on missions around the world. As part of a COVID-19 risk mitigation strategy, Belgian Armed Forces that engaged in missions and operations abroad were screened (7683 RT-qPCR tests), pre- and post-mission, for the presence of SARS-CoV-2, including the identification of viral lineages. Nine distinct viral genotypes were identified in soldiers returning from operations in Niger, the Democratic Republic of the Congo, Afghanistan, and Mali. The SARS-CoV-2 variants belonged to major clades 19B, 20A, and 20B (Nextstrain nomenclature), and included "variant of interest" B.1.525, "variant under monitoring" A.27, as well as lineages B.1.214, B.1, B.1.1.254, and A (pangolin nomenclature), some of which are internationally monitored due to the specific mutations they harbor. Through contact tracing and phylogenetic analysis, we show that isolation and testing policies implemented by the Belgian military command appear to have been successful in containing the influx and transmission of these distinct SARS-CoV-2 variants into military and civilian populations.
Assuntos
COVID-19/virologia , Militares , SARS-CoV-2/classificação , SARS-CoV-2/genética , Afeganistão/epidemiologia , Bélgica , COVID-19/epidemiologia , China/epidemiologia , República Democrática do Congo/epidemiologia , Genoma Viral , Genômica , Humanos , Mali/epidemiologia , Epidemiologia Molecular , Mutação , Níger/epidemiologia , Filogenia , Viagem , Sequenciamento Completo do GenomaRESUMO
Coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) compromises the ability of military forces to fulfill missions. At the beginning of May 2020, 22 out of 70 Belgian soldiers deployed to a military education and training center in Maradi, Niger, developed mild COVID-19 compatible symptoms. Immediately upon their return to Belgium, and two weeks later, all seventy soldiers were tested for SARS-CoV-2 RNA (RT-qPCR) and antibodies (two immunoassays). Nine soldiers had at least one positive COVID-19 diagnostic test result. Five of them exhibited COVID-19 symptoms (mainly anosmia, ageusia, and fever), while four were asymptomatic. In four soldiers, SARS-CoV-2 viral load was detected and the genomes were sequenced. Conventional and genomic epidemiological data suggest that these genomes have an African most recent common ancestor and that the Belgian military service men were infected through contact with locals. The medical military command implemented testing of all Belgian soldiers for SARS-CoV-2 viral load and antibodies, two to three days before their departure on a mission abroad or on the high seas, and for specific missions immediately upon their return in Belgium. Some military operational settings (e.g., training camps in austere environments and ships) were also equipped with mobile infectious disease (COVID-19) testing capacity.
Assuntos
Betacoronavirus/isolamento & purificação , Infecções por Coronavirus/epidemiologia , Militares/estatística & dados numéricos , Pneumonia Viral/epidemiologia , Adulto , Bélgica/epidemiologia , COVID-19 , Teste para COVID-19 , Técnicas de Laboratório Clínico , Infecções por Coronavirus/diagnóstico , Surtos de Doenças , Humanos , Masculino , Epidemiologia Molecular , Níger/epidemiologia , Pandemias , Pneumonia Viral/diagnóstico , Reação em Cadeia da Polimerase em Tempo Real/métodos , SARS-CoV-2 , Testes Sorológicos , Carga Viral , Adulto JovemRESUMO
Ticks are vectors for a broad range of pathogens of medical and veterinary importance, such as Borrelia spp., Babesia spp., Anaplasma spp., Rickettsia spp., Bartonella spp. and the tick-borne encephalitis virus. The Gram-negative bacterium Anaplasma phagocytophilum is present worldwide, including Belgium where numerous patients were shown to harbour antibodies against this pathogen as recorded by the Belgian National Reference Center (NRC) for Anaplasma. The clinical presentation of human granulocytic anaplasmosis is an acute, febrile, nonspecific, flu-like illness. Leukopenia, thrombocytopenia and increased hepatic transaminase activities are commonly present early in the disease. Diagnosis early in the course of infection relies on the detection of antibodies or of the bacterium in the blood, as is performed at the NRC for Anaplasma, part of the Clinical Laboratory of the Queen Astrid Military Hospital in Brussels, Belgium. In this article, we discuss diagnostic test results as well as recent clinical and demographic characteristics of patients whose samples were analyzed by the NRC for Anaplasma in a four-year period (2013-2016).
Assuntos
Anaplasma phagocytophilum , Anaplasmose , Técnicas de Laboratório Clínico/métodos , Vetores de Doenças , Anaplasma phagocytophilum/imunologia , Anaplasma phagocytophilum/isolamento & purificação , Anaplasmose/epidemiologia , Anaplasmose/imunologia , Anaplasmose/fisiopatologia , Anaplasmose/terapia , Animais , Anticorpos/análise , Bélgica/epidemiologia , Demografia , Gerenciamento Clínico , Feminino , Humanos , Masculino , Prevalência , Estações do Ano , Avaliação de Sintomas/métodos , CarrapatosRESUMO
BACKGROUND: The geographic distribution of Dermacentor reticulatus is expanding in Europe. Surveillance of this tick species and its pathogens is desirable, as it transmits pathogens of public and veterinary importance. A high-throughput real-time PCR-based array was used to screen 1.741 D. reticulatus ticks from Belgium, Germany, The Netherlands, and Great Britain for the presence of 28 tick-borne bacteria and twelve protozoan parasites. The presence of pathogen DNA was confirmed by conventional PCR followed by sequencing. RESULTS: The array detected the presence of DNA from Borrelia spp. (7%), B. afzelii (0.1%), B. garinii (0.1%), B. spielmanii (0.1%), B. miyamotoi (0.2%), Anaplasma marginale (0.1%), A. phagocytophilum (0.1%), Ehrlichia canis (2%), Rickettsia helvetica (0.2%), spotted fever group Rickettsia (9.6%), Francisella tularensis or Francisella-like endosymbionts (95%), Coxiella burnettii (0.1%), Babesia divergens (0.2%), B. canis (0.9%) B. vogeli (5.6%), and Theileria equi (0.1%). Only the presence of B. canis and spotted fever group Rickettsia could be confirmed by conventional PCR and sequencing. The spotted fever Rickettsia-positive samples were all identified as R. raoultii. CONCLUSIONS: We successfully detected and determined the prevalence of B. canis and R. raoultii in D. reticulatus. An high-throughput array that allows fast and comprehensive testing of tick-borne pathogens is advantageous for surveillance and future epidemiological studies. The importance of thorough validation of real-time PCR-based assays and careful interpretation is evident.
RESUMO
In this paper we aim to add additional knowledge regarding the occurrence, origin and epidemiological features of the English sweating sickness. The English sweating sickness raged in five devastating epidemics with mortality rates between 30 and 50% between 1485 and 1551 throughout England, and on one occasion also affected mainland Europe, in 1529. The Picardy sweat, generally considered as the English sweating sickness' lesser deadly successor, flared up in France in 1718 and caused 196 localized outbreaks with varying severity all over France and neighboring countries up to 1861. The English sweating sickness has been the subject of numerous attempts to define its origin, but so far all efforts have failed due to lack of material, DNA or RNA, that - using modern techniques and knowledge - could shed light on its cause. Although the time frame in which the English sweating sickness occurred and the geographical spread of the outbreaks is generally known, we will demonstrate here that there was more to it than meets the eye. We found reports of cases of sweating sickness in years before, after and between the 1485, 1508, 1517, 1529 and 1551 epidemics, as well as reports of sweating sickness in Italy and Spain. CONCLUSION: In spite of the fact that the English sweating sickness apparently has not caused casualties for a more than a century now, we suggest that -given the right circumstances- the possibility of re-emergence might still exist. The fact that up until today we have no indication concerning the causal pathogen of the English sweating sickness is certainly not re-assuring.
Assuntos
Doença do Suor/história , Surtos de Doenças , Inglaterra/epidemiologia , Europa (Continente)/epidemiologia , História do Século XV , História do Século XVI , HumanosRESUMO
To estimate the seroprevalence of Borrelia burgdorferi (Bb), Anaplasma phagocytophilum (Ap), and Francisella tularensis (Ft) in Belgium, we tested sera from three population-based samples in which exposure to pathogen is assumed to vary: 148 samples from workers professionally exposed, 209 samples from rural blood donors, and 193 samples from urban blood donors. Sera were tested using ELISA or the immunofluorescence assay test. The seroprevalence of Bb was 5.4% in workers professionally exposed, 2.9% in rural blood donors, and 2.6% in urban blood donors, which is similar to other studies. The fraction of negative results decreases significantly from urban blood donors and rural blood donors to workers. Regarding the seroprevalence of Ap, the cutoff titer of 1:64 recommended by the manufacturer may be set too low and produces artificially high seroprevalence rates. Using a cutoff titer of 1:128, the seroprevalence of Ap was estimated at 8.1% for workers professionally exposed, 6.2% for rural blood donors, and 5.7% for urban blood donors. Tularemia sera confirmed the presence of the pathogen in Belgium at 2.0% for workers and 0.5% for rural and urban blood donors. Our study is one of the few providing an estimation of the seroprevalences of Bb, Ap, and Ft in three different populations in Belgium, filling the gap in seroprevalence data among those groups. Our findings provide evidence that the entire Belgian population is exposed to Bb, Ap, and Ft infections, but a higher exposure is noticed for professionals at risk. Education on the risk factors for tick bites and preventive measures for both professionals exposed and the general population is needed.
Assuntos
Anaplasma phagocytophilum , Anaplasmose/epidemiologia , Borrelia burgdorferi , Francisella tularensis , Doença de Lyme/epidemiologia , Tularemia/epidemiologia , Adulto , Idoso , Anaplasmose/parasitologia , Bélgica/epidemiologia , Feminino , Humanos , Doença de Lyme/microbiologia , Masculino , Pessoa de Meia-Idade , Fatores de Risco , Estudos Soroepidemiológicos , Tularemia/microbiologiaRESUMO
An open, uncontrolled single centre study was conducted in the Travel Clinic at the Military Hospital, Brussels. Eighty-eight subjects were recruited who had a primary series of tick-borne encephalitis (TBE) vaccine more than 5 years ago and who never received a booster dose afterwards. Response rate after booster vaccination was very high: 84 out of 88 subjects (95.5%) had neutralizing antibodies on plaque reduction neutralization test and all (100%) had IgG antibodies on ELISA, on Day 21-28 after booster vaccination. This study adds valuable information to the common situation of delayed booster interval. The results of our study indicate that in young healthy travellers (<50 years), one booster vaccination after a primary series of TBE vaccine in the past is sufficient to obtain protective antibodies, even if primary vaccination is much longer than the recommended booster interval of 5 years.
Assuntos
Vírus da Encefalite Transmitidos por Carrapatos/imunologia , Encefalite Transmitida por Carrapatos/prevenção & controle , Imunização Secundária , Avaliação de Resultados em Cuidados de Saúde , Vacinas Virais/imunologia , Adulto , Bélgica , Ensaio de Imunoadsorção Enzimática , Feminino , Hospitais Militares , Humanos , Esquemas de Imunização , Masculino , Pessoa de Meia-Idade , Medicina Militar , Testes de Neutralização , Medicina de ViagemRESUMO
The English sweating sickness caused five devastating epidemics between 1485 and 1551, England was hit hardest, but on one occasion also mainland Europe, with mortality rates between 30% and 50%. The Picardy sweat emerged about 150 years after the English sweat disappeared, in 1718, in France. It caused 196 localized outbreaks and apparently in its turn disappeared in 1861. Both diseases have been the subject of numerous attempts to define their origin, but so far all efforts were in vain. Although both diseases occurred in different time frames and were geographically not overlapping, a common denominator could be what we know today as hantavirus infections. This review aims to shed light on the characteristics of both diseases from contemporary as well as current knowledge and suggests hantavirus infection as the most likely cause for the English sweating sickness as well as for the Picardy sweat.
Assuntos
Surtos de Doenças/história , Infecções por Hantavirus/história , Infecções por Hantavirus/virologia , Orthohantavírus/isolamento & purificação , Doença do Suor/história , Doença do Suor/virologia , Inglaterra/epidemiologia , França/epidemiologia , Infecções por Hantavirus/epidemiologia , História do Século XVI , História do Século XVII , História Medieval , Humanos , Doença do Suor/epidemiologiaRESUMO
BACKGROUND: Although Ixodes spp. are the most common ticks in North-Western Europe, recent reports indicated an expanding geographical distribution of Dermacentor reticulatus in Western Europe. Recently, the establishment of a D. reticulatus population in Belgium was described. D. reticulatus is an important vector of canine and equine babesiosis and can transmit several Rickettsia species, Coxiella burnetii and tick-borne encephalitis virus (TBEV), whilst Ixodes spp. are vectors of pathogens causing babesiosis, borreliosis, anaplasmosis, rickettsiosis and TBEV. METHODS: A survey was conducted in 2008-2009 to investigate the presence of different tick species and associated pathogens on dogs and cats in Belgium. Ticks were collected from dogs and cats in 75 veterinary practices, selected by stratified randomization. All collected ticks were morphologically determined and analysed for the presence of Babesia spp., Borrelia spp., Anaplasma phagocytophilum and Rickettsia DNA. RESULTS: In total 2373 ticks were collected from 647 dogs and 506 cats. Ixodes ricinus (76.4%) and I. hexagonus (22.6%) were the predominant species. Rhipicephalus sanguineus (0.3%) and D. reticulatus (0.8%) were found in low numbers on dogs only. All dogs infested with R. sanguineus had a recent travel history, but D. reticulatus were collected from a dog without a history of travelling abroad. Of the collected Ixodes ticks, 19.5% were positive for A. phagocytophilum and 10.1% for Borrelia spp. (B. afzelii, B. garinii, B. burgdorferi s.s., B. lusitaniae, B. valaisiana and B. spielmanii). Rickettsia helvetica was found in 14.1% of Ixodes ticks. All Dermacentor ticks were negative for all the investigated pathogens, but one R. sanguineus tick was positive for Rickettsia massiliae. CONCLUSION: D. reticulatus was confirmed to be present as an indigenous parasite in Belgium. B. lusitaniae and R. helvetica were detected in ticks in Belgium for the first time.
Assuntos
Anaplasma phagocytophilum/isolamento & purificação , Babesia/isolamento & purificação , Borrelia/isolamento & purificação , Rickettsia/isolamento & purificação , Carrapatos/classificação , Animais , Bélgica/epidemiologia , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães , Feminino , Masculino , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologia , Carrapatos/microbiologia , Carrapatos/parasitologiaRESUMO
The transmission of pathogens to susceptible hosts is dependent on the vector population dynamics. In Europe, bank voles (Myodes glareolus) carry Puumala hantavirus, which causes nephropathia epidemica (NE) in humans. Fluctuations in bank vole populations and epidemics in humans are correlated but the main factors influencing this relationship remain unclear. In Belgium, more NE cases are reported in spring than in autumn. There is also a higher incidence of human infections during years of large vole populations. This study aimed to better understand the link between virus prevalence in the vector, vole demography, habitat quality, and human infections. Three rodent populations in different habitats bordering Brussels city, Belgium, were studied for two years. The seroprevalence in voles was influenced first by season (higher in spring), then by vole density, vole weight (a proxy for age), and capture site but not by year or sex. Moreover, voles with large maximal distance between two captures had a high probability for Puumala seropositivity. Additionally, the local vole density showed similar temporal variations as the number of NE cases in Belgium. These results showed that, while season was the main factor influencing vole seroprevalence, it was not sufficient to explain human risks. Indeed, vole density and weight, as well as the local habitat, were essential to understanding the interactions in these host-pathogen dynamics. This can, in turn, be of importance for assessing the human risks.
Assuntos
Virus Puumala/patogenicidade , Animais , Arvicolinae/virologia , Bélgica , Febre Hemorrágica com Síndrome Renal/transmissão , Febre Hemorrágica com Síndrome Renal/virologia , Humanos , Análise Espaço-TemporalRESUMO
Hantavirus-specific serology tests are the main diagnostic technique for detection of hantavirus infection in Bosnia and Herzegovina. In order to enhance hantavirus infections monitoring a sensitive PCR based assay was developed to detect Dobrava (DOBV) and Puumala (PUUV) hantaviruses. Nested primer sets were designed within three different regions of the viral RNA (S and M segment of DOBV and M segment of PUUV) based on highly similar regions from a number of different European hantavirus strains. Assay conditions were optimized using cell cultures infected with DOBV Slovenia, PUUV Sotkamo and PUUV CG 18-20. This sensitive and specific assay has proven to be useful for detection of both Puumala and Dobrava hantaviruses.
Assuntos
Técnicas de Laboratório Clínico/métodos , Infecções por Hantavirus/diagnóstico , Técnicas de Diagnóstico Molecular/métodos , Orthohantavírus/isolamento & purificação , Reação em Cadeia da Polimerase/métodos , Virus Puumala/isolamento & purificação , Virologia/métodos , Animais , Bósnia e Herzegóvina , Chlorocebus aethiops , Primers do DNA/genética , Orthohantavírus/genética , Infecções por Hantavirus/virologia , Humanos , Virus Puumala/genética , RNA Viral/genética , Sensibilidade e Especificidade , Células VeroRESUMO
In Europe, hantaviruses (Bunyaviridae) are small mammal-associated zoonotic and emerging pathogens that can cause hemorrhagic fever with renal syndrome (HFRS). Puumala virus, the main etiological agent carried by the bank vole Myodes glareolus is responsible for a mild form of HFRS while Dobrava virus induces less frequent but more severe cases of HFRS. Since 2000 in Europe, more than 3000 cases of HFRS have been recorded, in average, each year, which is nearly double compared to the previous decade. In addition to this upside long-term trend, significant oscillations occur. Epidemic years appear, usually every 2-4 years, with an increased incidence, generally in localized hot spots. Moreover, the virus has been identified in new areas in the recent years. A great number of surveys have been carried out in order to assess the prevalence of the infection in the reservoir host and to identify links with different biotic and abiotic factors. The factors that drive the infections are related to the density and diversity of bank vole populations, prevalence of infection in the reservoir host, viral excretion in the environment, survival of the virus outside its host, and human behavior, which affect the main transmission virus route through inhalation of infected rodent excreta. At the scale of a rodent population, the prevalence of the infection increases with the age of the individuals but also other parameters, such as sex and genetic variability, interfere. The contamination of the environment may be correlated to the number of newly infected rodents, which heavily excrete the virus. The interactions between these different parameters add to the complexity of the situation and explain the absence of reliable tools to predict epidemics. In this review, the factors that drive the epidemics of hantaviruses in Middle Europe are discussed through a panorama of the epidemiological situation in Belgium, France, and Germany.
RESUMO
BACKGROUND: Ticks are the most important pathogen vectors in Europe. They are known to be influenced by environmental factors, but these links are usually studied at specific temporal or spatial scales. Focusing on Ixodes ricinus in Belgium, we attempt to bridge the gap between current "single-sided" studies that focus on temporal or spatial variation only. Here, spatial and temporal patterns of ticks are modelled together. METHODS: A multi-level analysis of the Ixodes ricinus patterns in Belgium was performed. Joint effects of weather, habitat quality and hunting on field sampled tick abundance were examined at two levels, namely, sampling level, which is associated with temporal dynamics, and site level, which is related to spatial dynamics. Independent variables were collected from standard weather station records, game management data and remote sensing-based land cover data. RESULTS: At sampling level, only a marginally significant effect of daily relative humidity and temperature on the abundance of questing nymphs was identified. Average wind speed of seven days prior to the sampling day was found important to both questing nymphs and adults. At site level, a group of landscape-level forest fragmentation indices were highlighted for both questing nymph and adult abundance, including the nearest-neighbour distance, the shape and the aggregation level of forest patches. No cross-level effects or spatial autocorrelation were found. CONCLUSIONS: Nymphal and adult ticks responded differently to environmental variables at different spatial and temporal scales. Our results can advise spatio-temporal extents of environment data collection for continuing empirical investigations and potential parameters for biological tick models.
Assuntos
Ecossistema , Ixodes/fisiologia , Animais , Comportamento Animal , Bélgica , Ninfa , Dinâmica Populacional , Análise de Regressão , Temperatura , Fatores de Tempo , Tempo (Meteorologia)RESUMO
Human granulocytic anaplasmosis (HGA) is a tick-borne rickettsial infection of neutrophils caused by Anaplasma phagocytophilum. Although the pathogen was known as a veterinary agent as early as 1932, the link with human disease was first established in 1990. In the past decennium, the involvement of HGA as an important and frequent cause of fever with a history of tick bite was increasingly recognized in many regions of Europe. This paper presents a 10-year A. phagocytophilum serosurveillance (2000-2009), wherein 1672 serum samples were tested and 418 were found positive. A total of 111 patients had a history of tick bite, fever, and at least a 4-fold rise in titre and are thus considered to be confirmed cases. These findings suggest that Belgium is a hot spot for HGA infections.
Assuntos
Anaplasma phagocytophilum/crescimento & desenvolvimento , Anaplasmose/sangue , Anticorpos Antibacterianos/sangue , Vetores Aracnídeos/classificação , Ixodes/classificação , Vigilância da População/métodos , Adolescente , Adulto , Idoso , Anaplasmose/complicações , Anaplasmose/diagnóstico , Anaplasmose/epidemiologia , Anaplasmose/imunologia , Anaplasmose/microbiologia , Anaplasmose/transmissão , Animais , Vetores Aracnídeos/fisiologia , Bélgica , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Febre/etiologia , Humanos , Ixodes/fisiologia , Masculino , Pessoa de Meia-Idade , Estudos Prospectivos , Estudos Soroepidemiológicos , Testes SorológicosRESUMO
Ticks can transmit a variety of viruses, bacteria or parasites that can cause serious infections or conditions in humans and animals. While tick-borne diseases are becoming an increasing and serious problem in Europe, tick-borne diseases are also responsible for major depressions in livestock production and mortality in sub-Saharan Africa, Latin America and Asia. This review will focus on the most important circulating tick-transmitted pathogens in Europe (Borrelia spp., Anaplasma phagocytophilum, Babesia spp., tick-borne encephalitis virus, Rickettsia spp. and Crimean-Congo hemorrhagic fever virus).