Publications by authors named "Christos Haralambous"

11 Publications

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Impact assessment of COVID-19 non-pharmaceutical interventions in long term care facilities in Cyprus: Safety improvement strategy.

Saf Sci 2021 Nov 17;143:105415. Epub 2021 Jul 17.

Center of Excellence in Risk & Decision Sciences, European University Cyprus, Nicosia 2404, Cyprus.

The current COVID-19 crisis has changed our everyday lives almost in every aspect. Many people worldwide have died or hospitalised due to the severe impact of COVID-19 on the vulnerable population, and in particular to the elderly residents of long term care facilities (LTCF). The problem is amplified due to the fact that many of those occupants also suffer from comorbidities (e.g. respiratory and cardiovascular diseases, hypertension, etc.) and are therefore regarded as a susceptible host to severe COVID-19 disease. Impacts can be felt in the wider societal safety level. The aim of the present study is, therefore, to present the first National multimodal quality and safety improvement strategy plan for the LTCF in the Republic of Cyprus. The current program focused on the intensification of COVID-19 epidemiological surveillance, the promotion of educational training on best practises in infection control and prevention, and the implementation of additional non-pharmaceutical interventions (NPIs), according to the recommendations of ECDC (European Centre for Disease Prevention and Control) and WHO (World Health Organization). This innovative program fostered the interconnectivity and collaboration among the local authorities, academia and the local leaders of the LTCF. In addition, this program reinforced the importance of volunteerism and active participation of medical students in the National initiatives against the COVID-19 pandemic. The effectiveness of the adopted multimodal advanced care-safety planning program is appraised based on the reported new confirmed COVID-19 cases among LTCF healthcare workers and occupants, after the introducing and implementation of the selected NPIs. This multimodal strategy plan seems to be capable of reducing significantly the number of new cases of COVID-19 infections in LTCF and as a result, to also affect the residents' death number.
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http://dx.doi.org/10.1016/j.ssci.2021.105415DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8285258PMC
November 2021

Extensive Testing and Public Health Interventions for the Control of COVID-19 in the Republic of Cyprus between March and May 2020.

J Clin Med 2020 Nov 8;9(11). Epub 2020 Nov 8.

Limassol General Hospital, 3304 Limassol, Cyprus.

Coronavirus disease 2019 (COVID-19) has significantly affected the well-being of individuals worldwide. We herein describe the epidemiology of COVID-19 in the Republic of Cyprus during the first epidemic wave (9 March-3 May 2020). We analyzed surveillance data from laboratory-confirmed cases, including targeted testing and population screening. Statistical analyses included logistic regression. During the surveillance period, 64,136 tests (7322.3 per 100,000) were performed, 873 COVID-19 cases were diagnosed, and 20 deaths were reported (2.3%). Health-care workers (HCWs) represented 21.4% of cases. Overall, 19.1% of cases received hospital care and 3.7% required admission to Intensive Care Units. Male sex (adjusted Odds Ratio (aOR): 3.04; 95% Confidence Interval (CI): 1.97-4.69), increasing age (aOR: 1.56; 95%CI: 1.36-1.79), symptoms at diagnosis (aOR: 6.05; 95%CI: 3.18-11.50), and underlying health conditions (aOR: 2.08; 95%CI: 1.31-3.31) were associated with hospitalization. For recovered cases, the median time from first to last second negative test was 21 days. Overall, 119 primary cases reported 616 close contacts, yielding a pooled secondary attack rate of 12% (95%CI: 9.6-14.8%). Three population-based screening projects, and two projects targeting employees and HCWs, involving 25,496 people, revealed 60 positive individuals (0.2%). Early implementation of interventions with targeted and expanded testing facilitated prompt outbreak control on the island.
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http://dx.doi.org/10.3390/jcm9113598DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC7695263PMC
November 2020

Genetic diversity and structure in Leishmania infantum populations from southeastern Europe revealed by microsatellite analysis.

Parasit Vectors 2013 Dec 5;6:342. Epub 2013 Dec 5.

Laboratory of Molecular Parasitology, Hellenic Pasteur Institute, 127 Vasilissis Sofias Avenus, 11521, Athens, Greece.

Background: The dynamic re-emergence of visceral leishmaniasis (VL) in south Europe and the northward shift to Leishmania-free European countries are well-documented. However, the epidemiology of VL due to Leishmania infantum in southeastern (SE) Europe and the Balkans is inadequately examined. Herein, we aim to re-evaluate and compare the population structure of L. infantum in SE and southwestern (SW) Europe.

Methods: Leishmania strains collected from humans and canines in Turkey, Cyprus, Bulgaria, Greece, Albania and Croatia, were characterized by the K26-PCR assay and multilocus enzyme electrophoresis (MLEE). Genetic diversity was assessed by multilocus microsatellite typing (MLMT) and MLM Types were analyzed by model- and distance- based algorithms to infer the population structure of 128 L. infantum strains.

Results: L. infantum MON-1 was found predominant in SE Europe, whilst 16.8% of strains were MON-98. Distinct genetic populations revealed clear differentiation between SE and SW European strains. Interestingly, Cypriot canine isolates were genetically isolated and formed a monophyletic group, suggesting the constitution of a clonal MON-1 population circulating among dogs. In contrast, two highly heterogeneous populations enclosed all MON-1 and MON-98 strains from the other SE European countries. Structure sub-clustering, phylogenetic and Splitstree analysis also revealed two distinct Croatian subpopulations. A mosaic of evolutionary effects resulted in consecutive sub-structuring, which indicated substantial differentiation and gene flow among strains of both zymodemes.

Conclusions: This is the first population genetic study of L. infantum in SE Europe and the Balkans. Our findings demonstrate the differentiation between SE and SW European strains; revealing the partition of Croatian strains between these populations and the genetic isolation of Cypriot strains. This mirrors the geographic position of Croatia located in central Europe and the natural isolation of the island of Cyprus. We have analysed the largest number of MON-98 strains so far. Our results indicate extensive gene flow, recombination and no differentiation between MON-1 and MON-98 zymodemes. No correlation either to host specificity or place and year of strain isolation was identified. Our findings may be associated with intensive host migration and common eco-epidemiological characteristics in these countries and give valuable insight into the dynamics of VL.
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http://dx.doi.org/10.1186/1756-3305-6-342DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4029556PMC
December 2013

Multilocus microsatellite typing (MLMT) of strains from Turkey and Cyprus reveals a novel monophyletic L. donovani sensu lato group.

PLoS Negl Trop Dis 2012 14;6(2):e1507. Epub 2012 Feb 14.

Laboratory of Molecular Parasitology, Hellenic Pasteur Institute, Athens, Greece.

Background: New foci of human CL caused by strains of the Leishmania donovani (L. donovani) complex have been recently described in Cyprus and the Çukurova region in Turkey (L. infantum) situated 150 km north of Cyprus. Cypriot strains were typed by Multilocus Enzyme Electrophoresis (MLEE) using the Montpellier (MON) system as L. donovani zymodeme MON-37. However, multilocus microsatellite typing (MLMT) has shown that this zymodeme is paraphyletic; composed of distantly related genetic subgroups of different geographical origin. Consequently the origin of the Cypriot strains remained enigmatic.

Methodology/principal Findings: The Cypriot strains were compared with a set of Turkish isolates obtained from a CL patient and sand fly vectors in south-east Turkey (Çukurova region; CUK strains) and from a VL patient in the south-west (Kuşadasi; EP59 strain). These Turkish strains were initially analyzed using the K26-PCR assay that discriminates MON-1 strains by their amplicon size. In line with previous DNA-based data, the strains were inferred to the L. donovani complex and characterized as non MON-1. For these strains MLEE typing revealed two novel zymodemes; L. donovani MON-309 (CUK strains) and MON-308 (EP59). A population genetic analysis of the Turkish isolates was performed using 14 hyper-variable microsatellite loci. The genotypic profiles of 68 previously analyzed L. donovani complex strains from major endemic regions were included for comparison. Population structures were inferred by combination of bayesian model-based and distance-based approaches. MLMT placed the Turkish and Cypriot strains in a subclade of a newly discovered, genetically distinct L. infantum monophyletic group, suggesting that the Cypriot strains may originate from Turkey.

Conclusion: The discovery of a genetically distinct L. infantum monophyletic group in the south-eastern Mediterranean stresses the importance of species genetic characterization towards better understanding, monitoring and controlling the spread of leishmaniasis in this region.
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http://dx.doi.org/10.1371/journal.pntd.0001507DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3279343PMC
June 2012

Leishmaniases and the Cyprus paradox.

Am J Trop Med Hyg 2010 Mar;82(3):441-8

Veterinary Services of Cyprus, Nicosia, Cyprus.

In Cyprus, leishmaniasis has been considered exclusively a veterinary problem. It was prevalent before 1945, and until its recent reemergence, it was nearly eradicated by 1996 as a consequence of the destruction of reservoir hosts and vectors. A survey carried out to provide an unbiased estimate of current transmission rates in dogs and humans showed a 9-fold increase in dog seroprevalence (reaching 14.9%) compared with 10 years ago. However, no human cases caused by Leishmania infantum were detected, although L. donovani cases were reported recently. The 62 strains isolated from dogs were typed as L. infantum MON-1 (98.4%), which is the predominating zymodeme in the Mediterranean region, and MON-98 (1.6%). The Phlebotomus species P. tobbi (vector of L. infantum in Cyprus), P. galilaeus, and P. papatasi were the predominant species captured. Two transmission cycles seem to run in parallel in Cyprus: in dogs with L. infantum and in humans with L. donovani.
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http://dx.doi.org/10.4269/ajtmh.2010.09-0282DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2829906PMC
March 2010

The paraphyletic composition of Leishmania donovani zymodeme MON-37 revealed by multilocus microsatellite typing.

Microbes Infect 2009 May-Jun;11(6-7):707-15. Epub 2009 Apr 17.

Institut für Mikrobiologie und Hygiene, Charité Universitätsmedizin Berlin, Berlin, Germany.

Multilocus microsatellite typing (MLMT) was employed to compare strains of Leishmania donovani belonging to the MON-37 zymodeme (MON-37 strains) from Cyprus and Israel to MON-37 strains from the Indian subcontinent, the Middle East, China and East Africa as well as strains of other zymodemes. The MLMT data were processed with a distance-based method for construction of phylogenetic trees, factorial correspondence analysis and a Bayesian model-based clustering algorithm. All three approaches assigned the MON-37 strains to different distantly related genetically defined subgroups, corresponding to their geographical origin. Specifically, the Kenyan, Sri Lankan and Indian MON-37 strains were genetically closer to strains of other zymodemes from the same regions than to MON-37 strains from other areas. MON-37 strains from Cyprus and Israel were clearly different not only among themselves, but also compared to all the other MON-37 strains studied and could, therefore, be autochthonous. This study showed that the zymodeme MON-37 is paraphyletic and does not reflect the genetic relationship between strains of different geographical origin.
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http://dx.doi.org/10.1016/j.micinf.2009.04.009DOI Listing
August 2009

Differentiation and gene flow among European populations of Leishmania infantum MON-1.

PLoS Negl Trop Dis 2008 Jul 9;2(7):e261. Epub 2008 Jul 9.

Institut für Mikrobiologie und Hygiene, Charité Universitätsmedizin Berlin, Berlin, Germany.

Background: Leishmania infantum is the causative agent of visceral and cutaneous leishmaniasis in the Mediterranean region, South America, and China. MON-1 L. infantum is the predominating zymodeme in all endemic regions, both in humans and dogs, the reservoir host. In order to answer important epidemiological questions it is essential to discriminate strains of MON-1.

Methodology/principal Findings: We have used a set of 14 microsatellite markers to analyse 141 strains of L. infantum mainly from Spain, Portugal, and Greece of which 107 strains were typed by MLEE as MON-1. The highly variable microsatellites have the potential to discriminate MON-1 strains from other L. infantum zymodemes and even within MON-1 strains. Model- and distance-based analysis detected a considerable amount of structure within European L. infantum. Two major monophyletic groups-MON-1 and non-MON-1-could be distinguished, with non-MON-1 being more polymorphic. Strains of MON-98, 77, and 108 were always part of the MON-1 group. Among MON-1, three geographically determined and genetically differentiated populations could be identified: (1) Greece; (2) Spain islands-Majorca/Ibiza; (3) mainland Portugal/Spain. All four populations showed a predominantly clonal structure; however, there are indications of occasional recombination events and gene flow even between MON-1 and non-MON-1. Sand fly vectors seem to play an important role in sustaining genetic diversity. No correlation was observed between Leishmania genotypes, host specificity, and clinical manifestation. In the case of relapse/re-infection, only re-infections by a strain with a different MLMT profile can be unequivocally identified, since not all strains have individual MLMT profiles.

Conclusion: In the present study for the first time several key epidemiological questions could be addressed for the MON-1 zymodeme, because of the high discriminatory power of microsatellite markers, thus creating a basis for further epidemiological investigations.
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http://dx.doi.org/10.1371/journal.pntd.0000261DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2438616PMC
July 2008

Development of a molecular assay specific for the Leishmania donovani complex that discriminates L. donovani/Leishmania infantum zymodemes: a useful tool for typing MON-1.

Diagn Microbiol Infect Dis 2008 Jan 21;60(1):33-42. Epub 2007 Sep 21.

Laboratory of Molecular Parasitology, Department of Microbiology, Hellenic Pasteur Institute, 115 21 Athens, Greece.

We have developed a simple, rapid, sensitive, and cost-effective typing method, based on the amplicon size of the K26 gene, capable of species/strain discrimination of Leishmania donovani complex strains causing visceral leishmaniasis (VL). It was evaluated on 112 strains and compared with multilocus enzyme electrophoresis (MLEE) typing. The K26 polymerase chain reaction (PCR) applied on 26 representative L. donovani complex strains gave 14 different amplicon sizes. The assay was specific to the L. donovani complex and discriminated L. infantum from L. donovani strains. MON-1 strains were also easily distinguished from other non-MON-1. Surprisingly, 29.3% of the Greek strains included in this study were MLEE typed as MON-98 and gave exclusively a 940-bp amplicon. The majority of Greek MON-1 strains gave also the 940-bp amplicon, whereas a 626-bp amplicon was consistently obtained with other European MON-1 strains. K26 PCR-restriction fragment length polymorphism, based on MON-1 K26 sequence polymorphism, gave 2 MON-1 subgroups. Application of the method may contribute to efficiently monitor VL.
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http://dx.doi.org/10.1016/j.diagmicrobio.2007.07.019DOI Listing
January 2008

Evolutionary and geographical history of the Leishmania donovani complex with a revision of current taxonomy.

Proc Natl Acad Sci U S A 2007 May 21;104(22):9375-80. Epub 2007 May 21.

*Biology Centre, Institute of Parasitology, Czech Academy of Sciences, and Faculty of Biology, University of South Bohemia, 370 05 Ceské Budejovice, Czech Republic.

Leishmaniasis is a geographically widespread severe disease, with an increasing incidence of two million cases per year and 350 million people from 88 countries at risk. The causative agents are species of Leishmania, a protozoan flagellate. Visceral leishmaniasis, the most severe form of the disease, lethal if untreated, is caused by species of the Leishmania donovani complex. These species are morphologically indistinguishable but have been identified by molecular methods, predominantly multilocus enzyme electrophoresis. We have conducted a multifactorial genetic analysis that includes DNA sequences of protein-coding genes as well as noncoding segments, microsatellites, restriction-fragment length polymorphisms, and randomly amplified polymorphic DNAs, for a total of approximately 18,000 characters for each of 25 geographically representative strains. Genotype is strongly correlated with geographical (continental) origin, but not with current taxonomy or clinical outcome. We propose a new taxonomy, in which Leishmania infantum and L. donovani are the only recognized species of the L. donovani complex, and we present an evolutionary hypothesis for the origin and dispersal of the species. The genus Leishmania may have originated in South America, but diversified after migration into Asia. L. donovani and L. infantum diverged approximately 1 Mya, with further divergence of infraspecific genetic groups between 0.4 and 0.8 Mya. The prevailing mode of reproduction is clonal, but there is evidence of genetic exchange between strains, particularly in Africa.
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http://dx.doi.org/10.1073/pnas.0703678104DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC1890502PMC
May 2007
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