Sandfly surveillance and investigation of Leishmania spp. DNA in sandflies in Kosovo.

In the past decade, leishmaniasis seems to be re-emerging in Balkan countries. There are serious implications that Kosovo is a visceral leishmaniasis endemic region with autochthonous transmission; nevertheless, surveillance of vectors, reservoirs or the disease is not yet established. Gaining knowledge about sandfly vector species is a prerequisite for the development of a monitoring and control plan in the future. After a long gap in research of over 70 years, sandfly studies in Kosovo were resumed in 2014. During this presence/absence study, nine sandfly species were detected: Phlebotomus papatasi, Ph. perfiliewi, Ph. tobbi, Ph. neglectus, Ph. simici, Ph. balcanicus, Ph. alexandri, Ph. mascittii and Sergentomyia minuta. Three species are new with regard to the fauna of Kosovo - Ph. alexandri, Ph. balcanicus and Ph. mascittii. Besides increased diversity, changes in the number of collected specimens and distribution range of species were recorded, with Ph. neglectus being the most dominant species with the widest distribution. Testing of randomly chosen females for Leishmania spp. DNA resulted the in detection of L. tropica in a specimen of Ph. neglectus. The presence of numerous vector species in the sandfly fauna of Kosovo pose a threat for the re-emergence of vector-borne diseases. Therefore, continuous surveillance is recommended with regular updates on vector distribution and abundance.

Genetic diversity and population structure of Culex modestus across Europe: does recent appearance in the United Kingdom reveal a tendency for geographical spread?

In mainland Europe, the mosquito species Culex modestus Ficalbi (1890) is a bridge vector for West Nile virus (WNV) from its natural bird-mosquito cycle to mammals. The present study assessed the genetic diversity of Cx. modestus, as well as related Culex species, using the mitochondrial COI DNA barcoding region and compared this with the population structure across Europe. A haplotype network was mapped to determine genealogical relationships among specimens. The intraspecific genetic diversity within individual Culex species was below 2%, whereas the interspecific genetic divergence varied from 2.99% to 13.74%. In total, 76 haplotypes were identified among 198 sequences. A median-joining network determined from 198 COI sequences identified two major lineages that were separated by at least four mutation steps. A high level of intraspecific genetic diversity was not detected in Cx. modestus in samples submitted from different European populations, which indicates that morphologically identified specimens represent a single species and not a species complex. Therefore, it is deduced that different populations of Cx. modestus will show a similar potential to transmit WNV, lending support to concerns that the population present in southeast England represents a risk of transmission to humans.

Croatian guidelines for the pharmacotherapy of type 2 diabetes

INTRODUCTION: The Croatian Association for Diabetes and Metabolic Disorders of the Croatian Medical Association has issued in 2011 the first national guidelines for the nutrition, education, self-control, and pharmacotherapy of diabetes type 2. According to the increased number of available medicines and new evidence related to the effectiveness and safety of medicines already involved in the therapy there was a need for update of the existing guidelines for the pharmacotherapy of type 2 diabetes in the Republic of Croatia. PARTICIPANTS: as co-authors of the Guidelines there are listed all members of the Croatian Association for Diabetes and Metabolic Diseases, as well as other representatives of professional societies within the Croatian Medical Association, who have contributed with comments and suggestions to the development of the Guidelines. EVIDENCE: These guidelines are evidence-based, according to the GRADE system (eng. Grading of Recommendations, Assessment, Development and Evaluation), which describes the level of evidence and strength of recommendations. CONCLUSIONS: An individual patient approach based on physiological principles in blood glucose control is essential for diabetes' patients management. Glycemic targets and selection of the pharmacological agents should be tailored to the patient, taking into account the age, duration of disease, life expectancy, risk of hypoglyce- mia, comorbidities, developed vascular and other complications as well as other factors. Because of all this, is of national interest to have a practical, rational and applicable guidelines for the pharmacotherapy of type 2 diabetes.

Mass production cage for Aedes albopictus (Diptera:Culicidae).

Mass production is an important co mponent of any pest or vector control program that requires the release of large number of insects. As part of efforts to develop an area-wide program involving the sterile insect technique (SIT) for the control of mosquitoes, the Insect Pest Control Laboratory of the Food and Agriculture Organization-International Atomic Energy Agency (FAO-IAEA Joint Division) has developed a mass production cage (Aedes MPC) for brood stock colonies in a mass production system for Aedes albopictus (Skuse, 1895). A preliminary experiment using Plexiglas cages was carried out to estimate the impact of cage volume on egg productivity. Transparent Plexiglas cages of different dimensions but loaded with the same adult density were tested. Egg productivity (number of eggs laid per adult female) and adult survival were recorded and analyzed. According to the results, the optimal volume of 100 liters has been chosen to develop the Aedes MPC. The numbers of adults introduced into the Aedes MPC did not affect the egg production and adult survival in comparison with the Plexiglas cage experiment data, confirming the possible use of Aedes MPC for mass-rearing procedures. Finally, the modification of Aedes MPC and creation of a new prototype model of MPC (Anopheles MPC) to effectively contain Anopheles arabiensis (Patton, 1905) adults is discussed with major changes pioneered to oviposition devices and systems for automatic collection of the eggs.

Comparison of carbon dioxide, octenol and a host-odour as mosquito attractants in the Upper Rhine Valley, Germany.

Field studies were conducted in the Upper Rhine Valley to determine the responses of mosquitoes to CDC traps baited with either CO2, octenol, light or paired combinations of these. Among eight mosquito species caught, the attractant effect on trap catches was studied in the four most abundant: Aedes vexans, Ae.rossicus, Ae.cinereus and Culex pipiens. Traps baited only with light or octenol caught few mosquitoes, whereas many were caught by traps baited with CO2 alone or in combination with either of the other candidate attractants. CO2 baited traps, with or without light, caught the most Aedes. The combination of CO2 and octenol attracted most Cx pipiens, but this apparent synergy was not significant. Using a caged hamster compared to CO2 as bait in a CDC light-trap with only intermittent fan suction, the hamster attracted less mosquitoes than CO2 emitted at a rate of 225 g/h on days 1 and 2, whereas on days 3 and 4 the smell from the hamster's cage became significantly more attractive than this rate of CO2 for all species of mosquitoes.

Role of larval cadavers in recycling processes of Bacillus sphaericus.

The influence of larval cadavers of Culex pipiens on recycling processes of Bacillus sphaericus was investigated by bioassays and spore counts in the laboratory. Studies conducted with 3 different B. sphaericus concentrations (0.005, 0.01, 0.05 mg B. sphaericus/liter) indicated that the presence of cadavers in the water contributed to the maintenance of toxic levels of B. sphaericus. Larval cadavers seem to contain all the nutrients necessary both for vegetative multiplication and for toxin synthesis associated with the sporulation process. Bioassays of B. sphaericus revealed that the mortality of Culex pipiens remained on a high level over a period of 26 days when larval cadavers were added every second day to the test vessels. This result was supported by a sharp increase in spore density when cadavers were added at the same interval. The test series showed B. sphaericus recycles in intact cadavers of Culex pipiens, whereas this phenomenon could not be observed when crushed cadavers were used in the trials. Therefore, our results demonstrated that for successful recycling processes it seems of crucial importance that infected cadavers remain intact at least for a certain time and also that the dosage of the applied B. sphaericus plays a major role in recycling processes whereas larval density is only of minor importance to these processes.

Dependence of CO2-baited suction trap captures on temperature variations.

In order to provide better standardized CO2-baited trap samples, the relationship between sample size and evening temperatures was studied. Adult mosquitoes were collected for 3 h centered on sunset by an automatic interval suction trap baited with CO2 in the Upper Rhine Valley, Germany. Aedes vexans females were most abundant, and their blood-seeking activity showed a significant correlation with the evening's average temperature (r = 0.76, P < or = 0.05). A higher degree of correlation was obtained when the number of specimens caught was related to temperature indices (r = 0.93-0.98, P < or = 0.01). A regression equation indicates the lower temperature threshold for Aedes vexans was between 9 and 10 degrees C. For each degree of increase in the average temperature index, it was estimated that a 5.5% greater number of mosquitoes would be trapped. An upper temperature threshold for this species was not observed at average temperatures < or = 23 degrees C. A relationship between catches of Aedes rossicus and Aedes cinereus and temperatures recorded at the site during the study was not found.

A convenient mosquito membrane feeding system.

A convenient, electronically controlled, in situ, membrane feeding system is described. Evaluation of the system for feeding single pair-mated Aedes aegypti with defibrinated, refrigerated pig blood indicated no significant difference from mouse-fed controls in rat of egg maturation, fecundity, or pupal yield. The feeding system is also suitable for use with substitute protein meals.

Factors influencing the activity of Bacillus thuringiensis var. israelensis treatments.

Environmental factors influence the effectiveness of microbial control agents in mosquito control programs. Four of these factors (water temperature, larval density, sunlight and the effect of associated filter feeders) were studied with Bacillus thuringiensis var. israelensis under laboratory and semifield conditions in Europe using different instars of Aedes vexans, Ae. aegypti and Culex pipiens. Bioassays conducted at a low temperature (5 degrees C) yielded 10-fold higher LC50 and LC90 values compared with those conducted at a high temperature (25 degrees C). The efficacy of B.t.i. decreased in a linear manner with increasing larval density. Sunlight can reduce the effectiveness of B.t.i. by several times. Competition in food intake by filter feeding Daphnia resulted in lower mortality of mosquito larvae after B.t.i. applications.