A cluster of four cases of cutaneous leishmaniasis by Leishmania donovani in Cyprus: a case series

Introduction: Leishmaniasis is endemic in more than 95 countries and is the only tropical/subtropical vector-borne disease that has been endemic in Southern Europe for decades. To the best of our knowledge, this is the first case of cutaneous leishmaniasis by Leishmania donovani in a child and the first cluster with adult cases reported in Europe. Case presentation: We describe a familial cluster of four cutaneous leishmaniasis cases among Greek Cypriots caused by L. donovani in a Paphos village, in Cyprus. A 6-year-old boy (Case number 1) had a persistent lesion in the left angle of his upper lip, a 60-year-old woman (Case number 2) presented with a 2cm-diameter glabella lesion on her forehead, a 60-year-old man (Case number 3) developed a lesion on his moustache area and a 40-year-old woman (Case number 4) had a lesion on her neck. In Case number 3 the lesion was self-cured; the other cases recovered after surgical resection followed by liposomal amphotericin B (Case numbers 1 and 4) or thermotherapy and liposomal amphotericin B (Case number 2). Conclusions: This familial cluster of cutaneous leishmaniasis, due to the anthroponotic L. donovani, shows that the sand fly species responsible for transmitting this parasite species is found in the area around the three neighbouring houses involved. The factors favourable for the survival, spread and contact of the vector with people could be assessed in this area for the establishment of preventative measures to safeguard public health

Canine tick-borne pathogens in Cyprus and a unique canine case of multiple co-infections

Canine tick-borne pathogens such as Ehrlichia canis and Hepatozoon canis are widespread in the Mediterranean basin but have never been reported or investigated in Cyprus. We describe herein the presence of canine tick-borne pathogens in three dogs with clinical signs compatible with vector-borne diseases from Paphos area of Cyprus. Molecular and phylogenetic analysis revealed the presence of E. canis, Anaplasma platys, H. canis, Babesia vogeli and Mycoplasma haemocanis in Cyprus. One dog co-infected with E. canis, H. canis, B. vogeli and M. haemocanis is, to the best of our knowledge, the first report of this multiple co-infection in dogs. The tick-borne pathogens reported in the current study should be considered in the differential diagnoses in dogs exposed to ticks in Cyprus

Association between canine leishmaniosis and Ehrlichia canis co-infection: a prospective case-control study

Abstract Background In the Mediterranean basin, Leishmania infantum is a major cause of disease in dogs, which are frequently co-infected with other vector-borne pathogens (VBP). However, the associations between dogs with clinical leishmaniosis (ClinL) and VBP co-infections have not been studied. We assessed the risk of VBP infections in dogs with ClinL and healthy controls. Methods We conducted a prospective case-control study of dogs with ClinL (positive qPCR and ELISA antibody for L. infantum on peripheral blood) and clinically healthy, ideally breed-, sex- and age-matched, control dogs (negative qPCR and ELISA antibody for L. infantum on peripheral blood) from Paphos, Cyprus. We obtained demographic data and all dogs underwent PCR on EDTA-blood extracted DNA for haemoplasma species, Ehrlichia/Anaplasma spp., Babesia spp., and Hepatozoon spp., with DNA sequencing to identify infecting species. We used logistic regression analysis and structural equation modelling (SEM) to evaluate the risk of VBP infections between ClinL cases and controls. Results From the 50 enrolled dogs with ClinL, DNA was detected in 24 (48%) for Hepatozoon spp., 14 (28%) for Mycoplasma haemocanis, 6 (12%) for Ehrlichia canis and 2 (4%) for Anaplasma platys. In the 92 enrolled control dogs, DNA was detected in 41 (45%) for Hepatozoon spp., 18 (20%) for M. haemocanis, 1 (1%) for E. canis and 3 (3%) for A. platys. No Babesia spp. or “Candidatus Mycoplasma haematoparvum” DNA was detected in any dog. No statistical differences were found between the ClinL and controls regarding age, sex, breed, lifestyle and use of ectoparasitic prevention. A significant association between ClinL and E. canis infection (OR = 12.4, 95% CI: 1.5–106.0, P = 0.022) was found compared to controls by multivariate logistic regression. This association was confirmed using SEM, which further identified that younger dogs were more likely to be infected with each of Hepatozoon spp. and M. haemocanis, and dogs with Hepatozoon spp. were more likely to be co-infected with M. haemocanis. Conclusions Dogs with ClinL are at a higher risk of co-infection with E. canis than clinically healthy dogs. We recommend that dogs diagnosed with ClinL should be tested for E. canis co-infection using PCR

Towards Truly “Global” Near Infrared Calibrations for Protein and Neutral Detergent Fibre in Dried Ground Forages

Over the past five years, Foss and DeLaval have sponsored the activities of a group of forage analysts with the aim of developing global Near Infrared (NIR) calibrations for parameters that are important in ruminant nutrition. The approach adopted has been based on the amalgamation of historical databases from centres worldwide and calibrations for protein and neutral detergent fibre (NDF) in dried ground forages have been developed based on databases that currently comprise approximately 30,000 records. Protein and NDF, while not the most important parameters in ruminant nutrition, were chosen for the initial calibration development exercise because of the amount of data available and because the methodologies adopted by different laboratories worldwide were relatively uniform. The aim was to create calibrations that would work for any forage type in any area of the world. Over the past two years, several trials have been carried out worldwide comparing the performance of global calibrations with the performance of locally developed calibrations for indigenous forages and based on reference values from local laboratories

Spread of Vector-borne Diseases and Neglect of Leishmaniasis, Europe

Exotic vector-borne diseases are gaining attention at the expense of leishmaniasis

One Health: The global challenge of epidemic and endemic leishmaniasis

'One Health' proposes the unification of medical and veterinary sciences with the establishment of collaborative ventures in clinical care, surveillance and control of cross-species disease, education, and research into disease pathogenesis, diagnosis, therapy and vaccination. The concept encompasses the human population, domestic animals and wildlife, and the impact that environmental changes ('environmental health') such as global warming will have on these populations. Visceral leishmaniasis is a perfect example of a small companion animal disease for which prevention and control might abolish or decrease the suffering of canine and human patients, and which aligns well with the One Health approach. In this review we discuss how surveillance for leishmaniases is undertaken globally through the control of anthroponootic visceral leishmaniasis (AVL) and zoonotic visceral leishmaniasis (ZVL). The ZVL epidemic has been managed to date by the culling of infected dogs, treatment of human cases and control of the sandfly vector by insecticidal treatment of human homes and the canine reservoir. Recently, preventive vaccination of dogs in Brazil has led to reduction in the incidence of the canine and human disease. Vaccination permits greater dog owner compliance with control measures than a culling programme. Another advance in disease control in Africa is provided by a surveillance programme that combines remote satellite sensing, ecological modelling, vector surveillance and geo-spatial mapping of the distribution of vectors and of the animal-to-animal or animal-to-human pathogen transmission. This coordinated programme generates advisory notices and alerts on emerging infectious disease outbreaks that may impede or avoid the spreading of visceral leishmaniasis to new areas of the planet as a consequence of global warming

Epidemiological investigation of leishmaniasis in Cyprus: control and surveillance of the disease

Leishmaniases are diseases caused by members of the genus Leishmania, protozoan parasites infecting numerous mammal species including humans, and transmitted by the bite of phlebotomine sandflies. In Cyprus, leishmaniasis has been considered exclusively a veterinary problem. The aim of the study is the epidemiological investigation of leishmaniases in Cyprus, in dogs and humans, for the control and surveillance of the disease. The survey conducted to resolve this was based on the assumption that if there were to be VL cases, they had to be in the areas with the highest dog seroprevalence. To locate these areas, an epidemiological study was carried out on the dog population. A map of Cyprus was divided into 82 equal squares of which 30 were chosen randomly as the study area. Dog seroprevalence revealed the areas with the highest risk for humans. In such two areas, as well as in one with zero dog seroprevalence, a seroepidemiological study was conducted on the human population and sandfly collections were done to compare species in these three areas. The Government controlled part of the island, southern Cyprus (Fig.1), covering 5,896 km2 and comprising five prefectures, was divided into 82 equal arbitrary squares on a map. The squares were given numbers and 30 numbers were drawn from a ballot to select the squares to be included in the study. One or more villages falling in each of the 30 squares, according to dog population size, were chosen so as to have at least 200 dogs per square. A total of 900 dogs (379 males and 521 females; 719 hunting, 87 guard, 94 companion) living in the 30 areas were sampled (30 dogs per area) during 2005 and 2006. Owned dogs, living permanently in the area irrespective of race, age color or health status were included in the study. To select the dogs, the houses in each village in the study area were given numbers and drawn from a ballot; visited with a government veterinarian and the dog owners were informed about leishmaniasis and asked to participate in the study by providing a written consent. The dogs were examined clinically, peripheral blood (900 samples) and lymph node aspirates when enlarged lymph nodes were observed (22 samples) were collected and personal, epidemiological and clinical data were registered in questionnaires for each dog. Severe leishmaniasis symptoms were diagnosed in 18 dogs, the disease was confirmed by serology and PCR, the dogs were euthanized with the consent of the owner and the spleen provided. In addition, 2,056 dog sera were provided by veterinarians throughout the island for routine testing for leishmaniasis. ...................Οι λεϊσμανιάσεις είναι μολυσματικές ασθένειες που μεταδίδονται μέσω αιματοφάγου διαβιβαστή ξενιστή, τη σκνίπα και είναι ανθρωπονόσοι ή/ ζωονόσοι. Στην Κύπρο, ένα νησί στο όριο της Ευρώπης με την Ασία και την Αφρική, η ΣΛ θεωρείτο μέχρι πρότινος ασθένεια των σκύλων, καθώς δεν υπήρχε επίσημη αναφορά ανθρώπινων περιστατικών μέχρι και το 2005. Σκοπός αυτής της διατριβής ήταν η επιδημιολογική διερεύνηση της λεϊσμανίασης στην Κύπρο, στο σκύλο και στον άνθρωπο, για τον έλεγχο και την επιτήρηση της νόσου και η ανάπτυξη ενός ικανού συστήματος ενεργού επιτήρησης της ασθένειας, με σκοπό το φραγμό μετάδοσης της στον άνθρωπο και την παρεμπόδιση της εξάπλωσης της στο σκύλο. Η έρευνα σχεδιάστηκε και εκτελέστηκε έτσι ώστε να εντοπιστούν οι περιοχές με το υψηλότερο ποσοστό κρουσμάτων στα σκυλιά, οι περιοχές δηλαδή με τον μεγαλύτερο κίνδυνο για τη μετάδοση της λεϊσμανίασης στον άνθρωπο και οι περιοχές με το χαμηλότερο ποσοστό ή αρνητικές, περιοχές δηλαδή με χαμηλό κίνδυνο για τη μετάδοση της λεϊσμανίασης στον άνθρωπο. Σε αυτές τις περιοχές πραγματοποιήθηκε οροεπιδεμιολογική έρευνα στον άνθρωπο και έγιναν συλλήψεις Φλεβοτόμων με σκοπό την σύγκριση των ειδών που παρατηρούνται στις τρεις περιοχές. Για τον σκοπό αυτό, οι ελεύθερες περιοχές της Κύπρου, έκτασης 5,896 τετραγωνικών χιλιόμετρων, χωρίστηκαν σε χάρτη σε 82 ίσα τετράγωνα. Στο κάθε τετράγωνο δόθηκε αριθμός 1-82 και με κλήρωση επιλέχθηκαν τυχαία 30 τετράγωνα. Μια περιοχή-τετράγωνο μπορούσε να περιλαμβάνει 1,2,3 ή και περισσότερα χωριά ή πόλεις. Από αυτά επιλέχθηκαν, με κλήρωση, τα χωριά στα οποία πραγματοποιήθηκαν οι δειγματοληψίες στα σκυλιά και η επιλογή του αριθμού των χωριών σε κάθε περίπτωση ήταν ανάλογη του πληθυσμού των σκυλιών ώστε να περιλαμβάνει τουλάχιστο 200 ζώα ανά τετράγωνο. Από σκυλιά που ζούσαν στα 30 τετράγωνα ελήφθησαν συνολικά δείγματα από 900 ζώα (30 σκυλιά ανά τετράγωνο). Για το σκοπό αυτό, οι οικίες που θα συμπεριλαμβάνονταν στη μελέτη, από τα χωριά των 30 τετραγώνων, επιλέγηκαν μετά από κλήρωση. Τα σκυλιά εξετάστηκαν κλινικά και ελήφθησαν δείγματα περιφερικού αίματος (900 δείγματα) και πολφός λεμφογαγγλοίων στις περιπτώσεις που διαπιστώθηκε διόγκωση των λεμφαδένων (22 δείγματα). Για κάθε σκυλί συμπληρώθηκε ερωτηματολόγιο με όλα τα προσωπικά, επιδημιολογικά και κλινικά του στοιχεία. Σοβαρά κλινικά συμπτώματα διαγνώστηκαν σε 18 σκυλιά, στα οποία έγινε ευθανασία μετά από επιβεβαίωση σπλαχνικής λεϊσμανίασης με ορολογικές και μοριακές εξετάσεις και προσκομίστηκε η σπλήνα για καλλιέργεια, μετά από τη σύμφωνη γνώμη του ιδιοκτήτη. Επιπρόσθετα εξετάστηκαν 2056 δείγματα ορών αίματος από σκυλιά από όλες τις ελεύθερες περιοχές της Κύπρου, που ελήφθησαν από ιδιώτες κτηνίατρους από ζώα για σκοπούς ελέγχου ρουτίνας. ..................

Geographical Distribution of MDR1 Expression in Leishmania Isolates, from Greece and Cyprus, Measured by the Rhodamine-123 Efflux Potential of the Isolates, Using Flow Cytometry

Leishmaniasis, a neglected vector-borne disease caused by the protozoan parasite Leishmania, is encountered in 98 countries causing serious concerns to public health. The most alarming is the development of parasite drug resistance, a phenomenon increasingly encountered in the field rendering chemotherapy ineffective. Although resistance to drugs is a complex phenomenon, the rate of efflux of the fluorescent dye Rhodamine-123 from the parasite body, using flow cytometry, is an indication of the isolate's ability to efflux the drug, thus avoiding death. The rate of efflux measured 275 Leishmania strains, isolated from patients and dogs from Greece and Cyprus, was measured and mapped to study the geographical distribution of the multidrug resistance (MDR) gene expression as an indication of the drug resistance of the parasite. The map showed that out of the seven prefectures, where dogs presented high efflux rates, five also had patients with high efflux rates. In one, out of the 59 prefectures studied, the highest number of isolates with efflux slope α > 1, in both human and dog isolates, was found; a fact which may suggest that spread of drug resistance is taking place. The virulence of the Leishmania strains, assessed after infecting human macrophages of the THP-1 cell line, fluctuated from 1% to 59.3% with only 2.5% of the isolates showing infectivity > 50%. The most virulent strains were isolated from Attica and Crete