According to entomological studies conducted over the past 30 years, there was low malaria transmission in suburb of Dakar but little evidence of it in the downtown area. However; there was some evidence of local transmission based on reports of malaria among permanent residents. An entomological evaluation of malaria transmission was conducted from May 2005 to October 2006 in two areas of Dakar.Plasmodium falciparum circumsporozoïte indexes were measured by ELISA, and the entomological inoculation rates (EIR) were calculated for both areas. Molecular assessments of pyrethroid knock down resistance (Kdr) and of insensitive acetylcholinesterase resistance were conducted.Mosquitoes were sampled by human landing collection during 34 nights in seven places in Bel-air area (238 person-nights) and during 24 nights in five places in Ouakam area (120 person-nights). Mosquitoes were identified morphologically and by molecular methods. The Anopheles gambiae s.l. respectively were caught in Bel-air and Ouakam. Three members of the complex were present: Anopheles arabiensis (> 98%), Anopheles melas (< 1%) and An. gambiae s.s. molecular form M (< 1%). Infected mosquitoes were caught only during the wintering period between September and November in both places. In 2005 and 2006, annual EIRs were 9,5 and 4, respectively, in Bel-air and 3 and 3, respectively, in Ouakam. The proportion of host-seeking An. gambiae s.l. captured indoors were 17% and 51% in Bel air and Ouakam, respectively. Ace 1 mutations were not identified in both members of the An. gambiae complex. Kdr mutation frequency in An. arabiensis was 12% in Bel-air and 9% in Ouakam.From May 2005 to October 2006, 4,117 and 797 An. melas in Dakar.Malaria is transmitted in Dakar downtown area. Infected mosquitoes were caught in two subsequent years during the wintering period in two distant quarters of Dakar. These data agree with clinical data from a Senegalese military Hospital of Dakar  where most malaria cases occurred between October and December. It was the first detection of  Plasmodium falciparum by Anopheles gambiae s.l. in Dakar were reported in different studies [Anopheles arabiensis was the main vector in this area [An. arabiensis as the vector [An. arabiensis were caught, and none of them were infected by P. falciparum [intra-muros (i.e. in the down-town area) and that the infections occurred in the suburbs or inland. Nevertheless, human malaria cases were reported in autochthonous people who had not been outside Dakar for at least one year, suggesting that malaria was transmitted in Dakar [In the sixties, the relative seriousness of malaria and the seasonal transmission of  studies -4. Immun studies . Malaria studies . In the his area . In the e vector . During lciparum -10. Base2.Located at 14°40'20" North, 17°25'22" West (the westernmost point of Africa), Dakar, the capital city of Senegal, has 1,030,594 inhabitants and covers the major part of the Cap Vert Peninsula. The altitude does not exceed 104 m. The population of the Dakar area is estimated to be 2.45 million people, representing 20% of the Senegalese population. The estimated population density is 12,233 inhabitants/kmThe study was conducted in two districts of Dakar: Bel-air in the east of the city and Ouakam in the west Figure . Bel-airThe Cap Vert Peninsula is located in the Atlantic Sudan zone. Two distinct seasons exist: a hot and wet season from June to November (maximum average temperature 28.2°C in October) and a cool and dry season from December to May (minimum average temperature 20.4°C in February). The first rains generally occur at the end of June or the beginning of July, and the last occur at the beginning of October. In 2005 and 2006 (the period covered by the study), the average rainfall was 525 and 350 mm, respectively.Sampling by human landing of malaria vectors was carried out both indoors and outdoors. Collectors gave prior informed consent and received anti-malaria prophylaxis and yellow fever immunization. Collectors were organized in teams of two for each collection point. Replacement of workers within a team was done every two hours from 7:00 PM to 7:00 AM. Teams of collectors were rotated among the collection points on different collection nights to minimize sampling bias. Landing catches were performed at seven points (two places indoors and five outdoors) in two periods, from May 2005 to May 2006 and from September to October 2006, during 34 nights in Bel-air (i.e. 238 person-nights). In Ouakam, human landing collections were performed at five points (two indoor and three outdoor locations) for two periods, from July 2005 to May 2006 and from September to October 2006, during 24 nights (i.e. 120 person-nights).et al [Mosquitoes were recorded by the location and hours of capture. They were sorted by genera, and anopheline mosquitoes were identified morphologically following the Gillies and Coetzee keys and by software from Hervy et al ,12. Culiet al . All mosP. falciparum circumsporozoite protein (CSP)[A. gambiae complex caught during the dry season, a random sample of females caught during the rainy season, together with all CSP-positive anopheline, were identified by polymerase chain reaction (PCR) at the species and molecular forms levels [Kdr and Ace1 mutations were carried out on these mosquitoes as previously described [Heads and thoraces of anopheline females were tested by enzyme-linked immunosorbent assay (ELISA) for ein (CSP). All fems levels . Moleculescribed ,17.An. gambiae s.l. biting activity is sufficient for transmission only during the end of the rainy period; the EIR calculated for this period will be considered as the annual EIR. Conformity of Kdr and Ace1 frequencies with Hardy-Weinberg expectations was tested using a Pearson chi-square test considered significant when P < 0.05. Endophagic rates were compared using a chi-square test.The human biting rate (HBR) was expressed as the number of female anopheline bites per human per night. The CSP index was calculated as the proportion of mosquitoes found to be positive for CSP. The entomological inoculation rate (EIR) was calculated as the product of the HBR and the CSP index of mosquitoes collected on humans. The Rainfall data were graciously provided by the National Weather Agency.Culex quinquefasciatus, 13.9% Culex tritaeniorynchus, 7.1% An. gambiae s.l., 2.2% Aedes aegypti; Tables A total of 69,082 mosquitoes were caught  were caught between July and December. The peak of biting occurred in September and October, at the end of the winter period (rainy season): 67% and 87% of mosquitoes were caught during these two months in Ouakam and Bel-air, respectively. During this period in 2005, in Bel-air, the average biting rate for An. gambiae s.l. was 112 bites per person per night, with a peak of 181 bites per person per night; in Ouakam, the average biting rate for An. gambiae s.l. was 19.7 bites per person per night, with a peak of 37.2 bites per person per night. During September-October in 2006, the average rates in Bel-air and Ouakam were 19.1 and 10.4 An. gambiae s.l. bites per person per night, respectively. The distribution of An. gambiae s.l. bites by hour is shown in Figure The biting activity at the two sites from May 2005 to May 2006 is shown in Figure An. gambiae s.l. caught indoor and outdoor were 121 and 115, respectively in Ouakam and 115 and 540, respectively in Bel air. The proportion of host-seeking An. gambiae s.l. captured indoors were 51% and 17% in Ouakam and Bel air, respectively. . From December 2005 to May 2006, none of the 16 mosquitoes were positive for CSP. In September and October 2006, three specimens among 1,068 were positive. The CSP index was 0.28% (CI95% = 0.06–0.82).In Bel-air, 3,049 Infected mosquitoes were identified only between the end of the rainy season. In 2005 and 2006, annual EIRs were 9.5 and 4 infective bites for a person without protection, respectively.An. gambiae s.l. collected by human landing catch were processed by ELISA for P. falciparum antigen detection in 2005, and 217 were processed in 2006. From July to the middle of September 2005, none of the 279 mosquitoes were positive. From the end of September to December, two specimens among 310 were positive. The CSP index was 0.64% (CI95% = 0.08–2.30). From December 2005 to May 2006, none of the 9 mosquitoes were positive. In September and October 2006, two specimens among 1,068 were positive. The CSP index was 0.96% (CI95% = 0.12–3.40). In 2005 and 2006, annual EIRs were 3 and 3 infective bites for a person without protection, respectively.In Ouakam, 589 An. gambiae s.l. captured before September 2005 , a random sample of the mosquitoes caught from September to December 2005 (346 from Bel-air and 216 from Ouakam), a random sample of the mosquitoes caught in September and October 2006 (137 from Bel-air and 134 from Ouakam), and all CSP-positive specimens were identified at the species level. In Bel-air, three members of the complex were present in 2005: An. arabiensis (97.8%), Anopheles melas (2.0%) and An. gambiae s.s. molecular form M (0.2%). In 2006, only An. arabiensis (98.5%) and An. gambiae s.s. molecular form M (1.5%) were present. In Ouakam, and only An. arabiensis was present in 2005. In 2006, one specimen of An. melas was caught. Emerging adults and all CSP-positive specimens were An. arabiensis.Using rDNA-PCR, all specimens of Kd-wr and Ace1 mutations. Ace 1 mutations were not identified in any members of the An. gambiae complex either in Ouakam or in Bel-air. The Kdr-w mutation frequency in An. arabiensis was 12% in Bel-air and 9% in Ouakam . The genotypic frequencies are shown in Table All CSP-positive mosquitoes per site and a random sample of 100 PCR-identified mosquitoes were tested for the P. falciparum-infected Anopheles have been caught during two rainy periods consecutively in two quarters of the town. All infected An. gambiae s.l. were caught only during the rainy period. The seasonal transmission occurs in the end of the rainy season from September to November. In August 2005, very heavy rains (278 mm of water in six days) flooded many parts of the city. Many potential breeding sites were created, and the An gambiae s.l. aggressiveness was high, with a peak of almost 200 bites per person per night in September. This abnormal situation in Dakar, due to an exceptional climatic event, could have led to an exceptional malaria transmission. However, the catch of infected Anopheles during the 2006 winter proved that malaria transmission is not exceptional and occurred each year during this period. These data agree with clinical data from a Senegalese military Hospital of Dakar , where most malaria cases occur between October and December.Malaria transmission is a reality in Dakar. An. arabiensis were infected. Anopheles arabiensis is the main member of the gambiae complex in Dakar and, according to its abundance and to the results of this study, is the main malaria vector. The presence of An. melas was detected in Dakar for the first time in the present study. Nevertheless, this vector was not found to be infected with P. falciparum. Until now, An. melas had been reported only in the mangrove swamps of the Delta's Saloum  and in the Senegal River delta  [Anopheles pharoensis nor An. gambiae molecular form M caught in Dakar were infected with P. falciparum. The aggressiveness of An. arabiensis was higher in Bel-air than in Ouakam, but EIRs were very high in the two quarters. An. arabiensis is more endophagic in Ouakam. This behavior allows for an easier access to a human blood meal that may explain the similar EIRs observed for the two quarters.Only specimens of Senegal) ,19. In aAn. gambiae s.l. aggressiveness lasts for a little longer than one month  are caught [An. arabiensis population, and to assess the gene flow between the urban and rural populations of Senegalese An. arabiensis.This situation questions the origins of these anopheline populations. Are there autochthonous populations from Dakar growing during the rainy season, or are they coming in from other areas of Senegal? Populations of y season . In Bark seasons . They coe caught . FurtherKdr) to pyrethroids and DDT of An. gambiae s.l is an increasing problem. Two mutations at the same locus in the voltage-gated sodium channel are known to confer knock-down resistance to a wide range of pyrethroids and DDT [Kdr-w and Kdr-e) in An. gambiae s.s. as well in An. arabiensis [An. arabiensis insecticide susceptibility in the area of Dakar. In 1987, bioassays were conducted in two places in the suburbs of Pikine and of Thiaroye [An. arabiensis was found in Dakar [Kdr-w frequency of 0.12 in Bel-air and 0.09 in Ouakam. The populations were not at Hardy-Weinberg equilibrium for Kdr in either site (P < 0.0001). This disequilibrium could be due to local selection pressure by the agricultural use of pyrethroids in market gardens.Pyrethroids are the main insecticide used in malaria vector control, including indoor residual spraying and impregnated materials . Pyrethroids have the advantage of acting very rapidly as insecticides, with both knockdown and lethal effects at dosages under the threshold of mammalian toxicity . Since 1 and DDT -37. Thesabiensis -41. TherThiaroye , where iin Dakar . Our datKdr mutation has been shown to be closely associated with DDT and pyrethroid resistance in several An. gambiae populations (particularly the molecular S form) [Kdr in conferring resistance in An. arabiensis remains unclear [An. arabiensis in Dakar. However, this is the first report of the Kdr-w mutation in An. arabiensis in Dakar. Resistance to carbamate and organophosphate insecticides is also widespread in West Africa. The presence of an insensitive acetylcholinesterase in populations of An. gambiae s.s. of both forms was revealed by biochemical assays. The ACE-1-R mutation has also been detected in the two molecular forms of An. gambiae s.s in many West African countries [An. arabiensis at present. Our results are consistent with these data.The  S form) -37. Howe unclear ,45 As a ountries ,46. ThisAn. arabiensis.In terms of effective vector control at the military camps and in town, choice of insecticide should depend on the results of susceptibility tests on P. falciparum isolates collected in Dakar [Plasmodium falciparum infections observed in Dakar during the rainy season. Are they due to autochthonous parasites transmitted during the winter or to parasites imported from the suburbs by commuters, as suggested by Vercruysse? [Malaria prevalence is very low in Dakar and its urban periphery ,8,10. Nein Dakar -50. Signcruysse?  In Senegcruysse? ,52.Urban malaria is considered to be an emerging problem in Africa. In 2003, 39% of Africa's people lived in urban settings; by 2030, 54% of Africans are expected to do so . With thAn melas and An gambiae s.s. in Dakar. Entomological studies are long, difficult and require time and special expertise but they are indispensable to understand the dynamics of malaria transmission in urban settlements and to monitor the increase of the insecticide resistance in urban mosquitoes. The study showed a difference in host finding behaviour between the two quarters: An. arabiensis are more endophagic in Ouakam than in Bel air. This difference of behaviour has an impact on the malaria transmission level. With a lower biting activity during the transmission season, the Annual EIR in Ouakam remains as high than in Bel air. Indoor vector control measures will probably not have the same impact in the two quarters. The misuse of impregnated bed nets during this period is probably riskier in Ouakam.The results of this study confirm that the vector complex situation in African towns is always changing, with the description of Malaria is transmitted in Dakar. This seasonal transmission occurs only during the two last months of the rainy season. The transmission level could be very high. Further studies have to be conducted in other parts of Dakar to assess the risk of transmission, to understand the role of permanent and temporary pools, the impact of urban farming and to discover the origin of the Dakar anopheline populations.The authors declare that they have no competing interests.FP was responsible for the study design, supervision of data collection, analysis, interpretation and production of the final manuscript and revisions. GT contributed to the supervision of data collection, the data analysis, interpretation and production of final manuscript. BP contributed to the supervision of data collection, to the data analysis, interpretation and production of final manuscript. LG contributed to the supervision of data collection, to the data analysis, interpretation and production of final manuscript. VM contributed to the data analysis and to the preparation of the final manuscript. FJ contributed to the supervision of data collection, to the data analysis. KP contributed to the supervision of data collection, to the data analysis. FB contributed to the supervision of data collection, to the data analysis. JFT was contributed to overall scientific management, analysis, interpretation and preparation of the final manuscript and revisions. CR was contributed to overall scientific management, analysis, interpretation and preparation of the final manuscript and revisions. CS was responsible for overall scientific management, analysis, interpretation and preparation of the final manuscript and revisions. All authors read and approved the final manuscript.


Malaria remains a major threat, to both travellers and military personnel deployed to endemic areas. The recommendations for travellers given by the World Health Organization is based on the incidence of malaria in an area and do not take the degree of exposure into account. The aim of this article is to evaluate the exposure of travellers by entomologic methods, which are the commonly used measures of the intensity of malaria transmission.From February 2004 to June 2004, five groups of 30 military personnel were stationed in up to 10 sites in western Côte d'Ivoire, from one week to several months. Adult mosquitoes were collected by human landing catches at each site during the five months and the level of exposure to malaria transmission of each group was estimated.The level of transmission varied from one site to another one from less than one to approximately more than 100 infective bites per month. In the majority of sites, at least two anopheline species were involved in transmission. The cumulative EIR over the study period varied according to the groups from 29 infected bites per person/per mission to 324.The level of malaria transmission and malaria risk varies widely (varying by a factor of eleven) between groups of travellers travelling in the same region and at the same time. Physicians involved in travel medicine or supporting expatriated populations or refugees should consider this heterogeneity and emphasize the importance of combining appropriate measures, such as chemoprophylaxis and protective measures against mosquitoes. Plasmodium sp. to anti-malarial drugs has led to an increasing interest in anti-vector protection measures [et al suggested that entomological inoculation rate (EIR) be used as a predictor of risk for non-immune persons [Malaria remains a major threat, both for travellers and for military personnel deployed each year to endemic areas. In France in 2007, up to 4,400 cases of malaria were estimated to have been imported, including 333 cases in military personnel . Chemoprmeasures . The maimeasures ,10. Seromeasures -15. For measures . Rombo e persons . However persons  has neve persons ,18. In f persons -24. ThesThe aim of this study was to estimate the level of exposure to malaria transmission of different groups of travellers with different travel itineraries during the same period in the same rural area in western Côte d'Ivoire by entomological methods.Plasmodium falciparum transmission area during the dry and rainy seasons [A company of 149 French soldiers, made up of five platoons of about 30 persons, was engaged in peace-keeping operations during a five-month period (from February to June 2004) in several rural areas in Western Côte d'Ivoire . Clinical malaria attacks were defined by the association of fever and Plasmodium infection. They were reported to the military epidemiological surveillance system.The soldiers used anti-malarial prophylaxis, including chemoprophylaxis of 100 mg of doxycycline daily, permethrin-impregnated battle dress, and deltamethrin-impregnated bed nets and repellents at night. The Army provided prophylactic drugs, anti-vectorial equipment and repellents. The military doctor for the company followed all subjects during their tour of duty and one month after their return. In the field, all fevers were recorded and Plasmodium infections were systematically investigated using rapid diagnostic test .This camp was situated at the eastern outskirts of Zoukougbeu village, bordered by the A6 main road going to Daloa.This is a small village surrounded by forest and cocoas plantations. The camp was situated at the northern outskirts of the village, near the entry, and was bordered by the path going north to Logouale and degraded forest.This is a big village on the northern side of Mont Peko that is surrounded by degraded forest and cocoa plantations. The camp was situated at the Western outskirts of the village and was bordered by a little stream, houses, and the path going to Dieouzon.The site called Danane was in fact situated two kilometers South of Danane. The camp was bordered by the A701 main road going from Danane to Bouenneu. Two villages were situated near (< 500 m) the camp.The site called Duekoue was in fact situated about three kilometers South of the Duekoue city, in a wood factory surrounded by forest and bordered by the A7 main road going to Guiglo.Anopheles gambiae s.l., Anopheles funestus s.l. and Anopheles nili s.l. by sites. Locally recruited collectors were organized into teams of two for each collection point. Workers in each team were switched out every hour from 6:00 PM to 7:00 AM. Teams of collectors were rotated among the collection points on different collection nights to minimize sampling bias. Collectors gave prior informed consent and received anti-malarial prophylaxis and yellow fever immunization.Adult mosquitoes were collected by human landing catches one to two nights per week at each site during the five-month study period. Sampling was carried out at four points (two indoors and two outdoors). The total person-nights of capture indoors and outdoors at each site during the mission are shown in additional file et al [The species of mosquitoes were recorded by day, site, and hours of capture. They were sorted by Genus and anopheline mosquitoes were identified morphologically following the keys of Gillies and Coetzee  and usinet al . Culicinet al . All mosPlasmodium falciparum was tested for using enzyme-linked immunosorbent assay (ELISA) [P. falciparum CSP, when there were less than 100 specimens or at least a random sample of 100 specimens by species and sorted by season (rainy or dry season).The presence of the circumsporozoite protein (CSP) of  (ELISA) ,30. All For each site and month of capture, all the specimens were analyzed for molecular identification if they were fewer than 100. When they were more than 100, a random sample of 100 specimens was considered for the molecular analysis. Moreover, all CSP positive specimens were identified by molecular methods.An. gambiae complex, the An. funestus and An. nili groups were identified by polymerase chain reaction (PCR) [Anopheles gambiae s.s. were molecularly distinguished into molecular forms [Females belonging to the on (PCR) -33 at thar forms . When M/ar forms ,35. DataAn. gambiae s.l., An. funestus s.l. and An. nili s.l. and their respective CSP rates.The human biting rate (HBR) was expressed as the average number of female anopheline bites per person per night during the dry season and rainy season. Specific CSP rates were calculated per species, per site and per season as the proportion of mosquitoes found to contain circumsporozoite antigen by ELISA. Ninety-five percent confidence intervals were calculated by the exact binomial method. Comparisons of these proportions were performed by Chi-square . Specific entomological inoculation rates (EIR) were calculated as the product of the HBR per site, per season for The malaria attack rate was calculated as the ratio of the number of clinical malaria cases to the number of soldiers participating in the mission.Anopheles, 20.8% were Culex, and 2.3% were Aedes, mainly Aedes aegypti (41.8%). In Logouale, the number of catching points was reduced to two (one indoors and one outdoors) after the first collection night due to the high number of Anopheles sp caught in this place. Anopheles were caught indoors (48% of captured specimens) as well as outdoors (52%). Of the 6,435 female anopheles collected, 65.2% were An. gambiae s.l., 19% An. funestus s.l. and 13.7% An. nili s.l. They were found at all sites, except in Zoukougbeu and Douekoue where An. nili was not caught. Other anopheline species were captured, including Anopheles pharoensis , Anopheles hancocki (mainly in Guezon), Anopheles dureni and Anopheles ziemanni.During the five-month period, a total of 12,520 female mosquitoes were caught during 81 collections nights (195 person-nights of capture) at ten different localities: 51.4% were An. gambiae s.l. collected, 1,626 were identified by PCR. They were all An. gambiae s.s. with An. gambiae s.s. M form (48.2%), M/S form (0.3%) and S form (51.5%). Out of the 1,219 An. funestus s.l. collected, 695 were identified by PCR, 99.1% were An. funestus s.s and 0.9% Anopheles leesoni . Out of the 880 An. nili s.l. collected, 396 were identified by PCR. They were all An. nili s.s.Out of the 4,197 An. gambiae s.s. captured at each permanent site, more than 90% in Bangolo and Guezon, 68% in Kahin, and 15% in Logouale were identified. Anopheles gambiae s.s., identified as the S molecular form, was most common in Guezon and Kahin, while the M molecular form was most common in Logouale . In Guezon, An. gambiae s.l. was exo-endophagic  and An. funestus s.l. was more endophagic (37% of bites occurred outdoors vs. 63% indoors. p < 0.05). In Kahin, An. gambiae s.l. and An. nili s.l. were exophagic and An. funestus s.l. was exo-endophagic. For An. gambiae s.l., 64% of bites occurred outdoors vs. 36% (p < 0.05). For An. nili s.l., 60% vs. 40% outdoors (p < 0.05), and for An. funestus s.l. 47% vs. 53% . In Logouale, An. gambiae s.l. and An. nili s.l. were exo-endophagic and An. funestus s.l. was exophagic. For An. gambiae s.l., 52% of bites occurred outdoors vs. 48% . For An. nili s.l., 42% vs. 58% , and for An. funestus s.l., 75% vs. 25% (p < 0.05).In Bangolo, An. gambiae s.l., An. funestus s.l. and An. nili s.l. bites per hour per site is shown in Figure An. gambiae s.l. and An. funestus s.l bites increased gradually between 7:00 p.m. and 1:00 a.m. reaching different peaks at each site. In Bangolo, the peak frequencies of bites were between 1:00 a.m. and 3:00 a.m., in Guezon between 4:00 a.m. and 5:00 a.m., and in Logouale and Kahin between 5:00 a.m. and 6:00 a.m.The distribution of An. nili s.l. bites was observed to increase gradually between 7:00 p.m. and 11:00 p.m. at all sites, and to peak between 11:00 p.m. and 00:00 a.m. and then decrease.The frequency of An. gambiae s.l. populations were found to be infected at both permanent sites and their CSP rates did not differ significantly between sites (p = 0.67). An. funestus s.l. populations were found to be infected at all sites except Logouale and their CSP rates did not differ significantly among sites (p = 0.36). Whatever the period, An. nili s.l, was found to be infected only in Kahin with a significant decrease between the dry and rainy period (p = 0.04). The CSP index for An. gambiae s.l. between the dry and the rainy season did not differ significantly. For An. funestus s.l., the CSP rates did not differ significantly between the dry and the rainy season expect in Kahin where it decreased in the rainy season (p = 0.03).Ninety of the 2,905 (3%) anopheline vectors were found to be positive by ELISA for circumsporozoite protein detection. The results by sites and by species are shown in Table An. gambiae s.l. caught during the five months at each site were identified at the species and molecular level and were all tested for CSP. None of the three M/S hybrids was infected. The difference between the CSP rates for An. gambiae M and S molecular form was not significant at any site. The CSP rates of An. gambiae M and S forms varied between the permanent sites from 1.75% to 10% . Five cases occurred in returnees: two cases in platoon 5 and 3 and one case in platoon 1. The malaria incidence rates were of 4.1% in Côte d'Ivoire and 3.4% after returning in France .The findings of this study clearly show that the level of malaria transmission and malaria risk varies widely (varying by a factor of eleven) between groups of travellers travelling in the same region and at the same time. Currently, the World Health Organization  and travAn. nili that thrive on the border of the river. This study confirms that, in wet savannah areas or in mountainous forested areas of Côte d'Ivoire, the rainy season, which occurs for most months in the year, allows the breeding sites to retain water much longer and thus sustain more substantial residual vector populations capable of ensuring malaria transmission even during the dry season [P. falciparum detection than the salivary glands detection that was used in previous studies.The level of transmission varied greatly from one site to another. These results are consistent with studies conducted in other parts of Africa that showed an important heterogeneity in malaria transmission between rural areas -43 and wy season -46. In ay season . Travel y season  and Man y season ,46, respy season -49, to gAn. gambiae s.s. and An. funestus s.s. were individually the only significant malaria vectors at three sites , in the other sites, the transmission was carried out by a vector association of at least two anopheline species, as previously described in Côte d'Ivoire [An. gambiae s.s. and An. funestus s.s,. with An. nili s.s. in a few places. In Guezon, An. funestus s.s. was the main vector during both dry and rainy seasons, and An. gambiae s.s was involved in transmission only during the rainy season. In Bangolo, An. gambiae s.s. was only found infected during the rainy season. The few human landing collections conducted here during the dry season did not allow P. falciparum-infected specimens to be caught, but An. gambiae s.s. was probably involved in malaria transmission in the dry season too. On the other hand, only a few specimens of An. funestus s.s., including one infected specimen, were caught in Bangolo, and these were only caught during the rainy season. Anopheles funestus s.s. was also only involved in malaria transmission as a secondary vector during the rainy season. In Logouale, An. gambiae s.s., An. funestus s.s. and An. nili s.s. were present during the dry and rainy seasons, but only An gambiae s.s. seemed significantly involved in malaria transmission. Indeed, the low An. funestus s.s. and An. nili s.s HBRs did not allow them to play a significant role in malaria transmission. In Kahin, An. gambiae s.s., An. funestus s.s. and An. nili s.s. were involved in malaria transmission during the dry and rainy seasons. Anopheles gambiae s.s was the main vector during the rainy season. Anopheles nili s.s was the main vector during the dry season. During this period, the EIR was above 60 infective bites by An. nili s.s. Anopheles nili breeding sites were probably located on the borders of the local river. During the wet season, the increase in the river water level probably destroyed most An. nili larval habitats. This is probably the explanation for the An. nili EIR and overall EIR decrease in Kahin during the rainy season. Others species were caught such as An. leesoni, An. hancocki, An. pharoensis and An. ziemanni, but none of them were found to be infected. During the study, in most of sites, the three dominant vectors bit humans as often indoors as they did outdoors.Although d'Ivoire ,50-52. TAn. nili s.l. bites occurred in the first part of the night. In Kahin, four of the ten infected An. nili were caught between 9.00 p.m. and midnight These findings  could limit the impact of insecticide-treated mosquito nets for local populations or travellers when they are exposed to An. nili bites. Furthermore, this study showed that 4% of bites occurred between 6:00 a.m. and 7:00 a.m. At the four permanent sites, three of the 27 infected An. funestus and 10 of the 53 infected An. gambiae were caught 6:00 a.m. and 7:00 a.m. Consequently, measures for anti-vector protection used in French military from 6:00 p.m. to 6:00 a.m., such as the wearing of long-sleeve clothes or the use of repellents, have been prolonged till 7:00 a.m. daily. These results showed that malaria transmission also occurs in the first hours of the day. Entomological studies often stopped the captures on 6:00 a.m., and, in some parts of Africa, they are probably underestimating the level of transmission.Most of et al in Cameroon [An. gambiae s.s. population composition between the dry and rainy seasons, as described by Della Torre et al in Burkina Faso [An. gambiae s.s. between western Côte d'Ivoire and Burkina Faso.The M form was predominant in sites where vegetation cover was deeper, like in Logouale and Douekoue. The S form was predominant in mountainous areas where forest cover was highly degraded (Guezon and Bleni Mehouin). In other places, the S-part of the population increased with anthropic modifications of the environment. This result is consistent with data from Wondji Cameroon . The varCameroon . Interesina Faso . This diAlthough exposure to malaria transmission is high, very few malaria cases occurred among the soldiers. Eleven cases were recorded: 54.5% of which occurred in Côte d'Ivoire and 45.5% after returning to France. These proportions are similar to those observed for the French Army from 1998 to 2006 . The attMalaria transmission in rural area is highly heterogeneous. The malaria exposure of travellers is highly dependent on this heterogeneity. Physicians involved in travel medicine or supporting expatriated population or refugees should emphasize the importance of vector control measures in protecting these populations (expatriated population or refugees) or counsel travellers before their stay in malaria transmission area. In addition, anti-vectorial measures are actually the only recommended measures for protection against vectors that transmit other pathogens, such as arboviruses.The authors declare that they have no competing interests.EOP: Contributed substantially to conception, study design, data collection, analysis and interpretation, preparation of the manuscript CR: conceived of the study, and participated in its design and coordination, data collection and critically reading the manuscript for important intellectual content. BK and CP: carried out data collection and trained the field staff. FJ, RG, VM and MB: processed entomological specimens in the laboratory, compiled the data and carried out statistical analyses. JPB: Participation in study design, coordination and critically reading of the manuscript. FP: Contributed substantially to conception, study design, data collection, analysis and interpretation, performed the statistical analysis and helped to draft the manuscript. All authors read and approved the final manuscript.This study was supported by Délégation Générale pour l'Armement  with the valuable assistance of the veterinary Chief Claire Dane.An. gambiae s.l., An. funestus s.l. and An. nili s.l. and number of species identified by molecular methodsBy site, HBR, CSP and monthly EIR of . The data provided in a table represent the results of human landing catches at each permanent and temporary sites realized from February to June 2004. The total Person-night of capture in dry season/rainy season (PNC), mean daily human biting rates (HBR), circumsporozoite protein rates (CSP) and entomological inoculation rates infective bites per person per month (EIR) of An. gambiae s.l., An. funestus s.l. and An. nili s.l. in each site are shown. The species identified by molecular methods and their number found in each site are presented too. 95% C.I.: 95% confidence interval.Click here for fileAn. gambiae s.l., An. funestus s.l. and An. nili s.l. from February to June 2004By permanent site, monthly HBR of . The data provided represent the mean daily human biting rate (HBR) of An. gambiae s.l., An. funestus s.l. and An. nili s.l. calculated by month, from February to June 2004, in four permanent sites. PNC: Total Person-night of capture.Click here for file


In recent years, human cyclosporiasis has emerged as an important infection, with large outbreaks in the United States and Canada. Understanding the biology and epidemiology of Cyclospora has been difficult and slow and has been complicated by not knowing the pathogen s origins, animal reservoirs (if any), and relationship to other coccidian parasites. This report provides morphologic and molecular characterization of three parasites isolated from primates and names each isolate: Cyclospora cercopitheci sp.n. for a species recovered from green monkeys, C. colobi sp.n. for a parasite from colobus monkeys, and C. papionis sp.n. for a species infecting baboons. These species, plus C. cayetanensis, which infects humans, increase to four the recognized species of Cyclospora infecting primates. These four species group homogeneously as a single branch intermediate between avian and mammalian Eimeria. Results of our analysis contribute toward clarification of the taxonomic position of Cyclospora and its relationship to other coccidian parasites.


He was born on 22nd September 1949. Dr. Gurwara passed his MBBS, DA and MS (Anaesth.) from M.L.N. Medical College, Allahabad, and joined M.L.B. Medical College, Jhansi, in 1976 as a lecturer. Thenceforth, on being promoted, he went on to become the head of the department and served as the head since July 2000 and retired from active service on 30th September 2009. He is survived by his wife, a son and a daughter. He was an active life member of ISA and was instrumental in the creation of Jhansi city branch being its first secretary and then president. He remained a patron of the branch till his end. He organised the annual conference of UP chapter ISA in 1987 and had been its Vice President twice.With profound grief, ISA, Jhansi city branch of UP chapter, informs the sad demise of Dr. A. K. Gurwara, Rtd. Professor and Head, Department of Anaesthesia, M.L.B. Medical College, Jhansi, on 10


Anopheles longipalpis is morphologically similar to the major African malaria vector Anopheles funestus at the adult stage although it is very different at the larval stage. Despite the development of the species-specific multiplex PCR assay for the An. funestus group, the genomic DNA of Anopheles longipalpis type C specimens can be amplified with the Anopheles vaneedeni and Anopheles parensis primers from this assay. The standard, species-specific An. funestus group PCR, results in the amplification of two fragments when An. longipalpis type C specimens are included in the analysis. This result can easily be misinterpreted as being a hybrid between An. vaneedeni and An. parensis. Anopheles longipalpis type C can be identified using a species-specific PCR assay but this assay is not reliable if other members of the An. funestus group, such as An. funestus, An. funestus-like and An. parensis, are included. The present study provides a multiplex assay that will identify An. longipalpis along with other common members of the African An. funestus group, including Anopheles leesoni.An. funestus, An. funestus-like, An. parensis, Anopheles rivulorum, An. vaneedeni and An. leesoni) in the An. funestus group and An. longipalpis type C from Malawi, Mozambique, South Africa and Zambia were used for the study. A restriction fragment length polymorphism (RFLP) assay was designed based on the DNA sequence information in the GenBank database.A total of 70 specimens from six species (EcoRI digested only An. longipalpis type C and An. funestus-like after the species-specific An. funestus group PCR assay. The An. longipalpis and An. funestus-like digestion profiles were characterized by three fragments, 376 bp, 252 bp and 211 bp for An. longipalpis type C and two fragments, 375 bp and 15 bp for An. funestus-like.The enzyme, An RFLP method for the group was developed that is more accurate and efficient than those used before. Hence, this assay would be useful for field-collected adult specimens to be identified routinely in malaria vector research and control studies. Anopheles funestus group originally consisted of nine species [An. funestus and eight minor or non-vectors, Anopheles aruni, Anopheles parensis, Anopheles vaneedeni, Anopheles confusus, Anopheles rivulorum, Anopheles leesoni, Anopheles brucei and Anopheles fuscivenosus. Subsequent studies on the systematics of the group have resulted in a reclassification of the group with An. funestus, An. aruni, An. parensis, An. confusus and An. vaneedeni being grouped together as members of the "An. funestus subgroup", An. rivulorum, An. rivulorum-like, An. brucei and An. fuscivenosus forming their own subgroup, and An. leesoni being classified with the Asian Anopheles minimus subgroup [An. funestus-like [An. funestus subgroup.The  species ,2: the msubgroup . Recentltus-like , and thiAnopheles longipalpis being one of these [An. funestus is very high. To further complicate the issue, molecular studies on this taxon have revealed at least two cryptic species, one from South Africa (Type A) and the other from Zambia (Type C) [In addition to the 10 species mentioned above, there are closely related species that are not included in the group because of morphological differences in the adult females - (Type C) .An. longipalpis has not been well studied despite the fact that it is widely distributed in eastern and southern Africa, from Sudan to Angola and South Africa [An. longipalpis from indoor and outdoor collections on Ukara Island in Lake Victoria and outdoors in Tanzania. Adugna and Petros [An. longipalpis specimens containing human blood meals from their collections in Ethiopia. Recently, Kent et al [An. funestus and Anopheles arabiensis in Zambia, it is predominantly zoophilic with only a small number feeding on humans. The species has never been implicated as a vector of malaria or involved in malaria transmission [The adult biology of h Africa ,2,6. Smih Africa ,8 reportd Petros  also fount et al  reportedsmission ,2,10,11.An. longipalpis are morphologically characterized by maxillary palpal length and pale basal and apical bands spanning the hind tarsal joints [An. funestus due to similarities of wing venation, palpal banding patterns and small body size. Currently, there are two identification methods for field-collected specimens of An. longipalpis. The first method, suggested by Kent et al [An. longipalpis using the species-specific PCR assay for the An. funestus group from the internal transcribed spacer (ITS) region of the rDNA [An. longipalpis type C which correlates with An. parensis and An. vaneedeni due to sequence similarity among these species. However, this analysis would lead to misidentification with hybrids of An. parensis and An. vaneedeni, although these have never been found in nature. The second method is a species-specific PCR assay for An. longipalpis from the ITS2 region developed by Koekemoer et al [An. funestus group are included such as An. funestus, An. funestus-like and An. parensis. Hence, this study proposes a new method for the molecular identification of An. longipalpis type C and six members of the An. funestus group using a Restriction Fragment Length Polymorphism (RFLP) that is more accurate and efficient than currently used methods.Although adult l joints ,2, this nt et al , is to ithe rDNA . The asser et al , but thiAn. longipalpis type C specimen available. Initially, specimens were identified morphologically using the keys of Gillies and Coetzee [An. longipalpis where the method of Collins et al [et al [An. funestus-like and the method of Koekemoer et al [Mosquitoes were collected from Malawi, Mozambique, South Africa and Zambia Table . There w Coetzee . All DNA Coetzee  except Ans et al  was usedl [et al  for An. er et al  for the et al [et al [An. funestus-like 390 bp fragment of Spillings et al [An. rivulorum 411 bp fragment of Koekemoer et al [EcoRI enzyme lies within the An. funestus-like and An. longipalpis type C fragments amplified by the methods of Spillings et al [et al [An. funestus-like and An. longipalpis type C directly. A total volume of 25 μL for each PCR reaction contained 1 μL of the genomic DNA of an individual mosquito, 1 × PCR Buffer, 0.2 mM of each dNTP, 0.26 pM  of each primer, and 1 unit of Taq DNA polymerase. The PCR cycling conditions were as follows: a 2 minute 94°C followed by 35 cycles of 30 seconds at 94°C, 30 seconds at 50°C and 40 seconds at 72°C; there was a final extension step of 10 minutes at 72°C. After amplification, 1 unit of EcoRI in 1 × buffer EcoRI (Roche Diagnostic) was added to the PCR reactions and digestion carried out at 37°C for a minimum of three hours. Digested fragments were electrophoresed through an ethidium bromide 2.5% agarose gel and photographed under ultraviolet light illumination using a gel imaging system.Although the method of Spillings et al  can be cl [et al , they regs et al  is closeAn. funestus group and An. longipalpis type C, resulted in different sizes of DNA fragments as recorded in Table An. longipalpis type C were amplified after the species-specific An. funestus group PCR. The restriction enzyme digested only the fragments for An. funestus-like and An. longipalpis type C. There were restriction sites at position 375 in An. funestus-like and at position 376 in the large fragment of An. longipalpis type C , An. funestus-like (375 bp) and An. rivulorum (411 bp) are close to each other. However, individuals that do not amplify at all may need additional processing using the An. longipalpis type A primers [The assay for the et al [An. longipalpis type C is similar to the sequences of An. parensis and An. vaneedeni in the An. funestus subgroup. These three species are almost identical at the adult stage with only a few minor differences [An. longipalpis was not included in the An. funestus group because of its very different larval morphology [et al [An. longipalpis type A, which is close to Anopheles pampanai and Anopheles varuna in the Asian An. minimus group based on the DNA analysis of the ITS2 region. They suggested that An. longipalpis type C should be placed in the An. funestus group supported by sequence similarity to both An. parensis and An. vaneedeni. Furthermore, Kent et al. [An. longipalpis type C were the same as the diagnostic amplicons for a hypothetical An. parensis/An. vaneedeni hybrid although no such hybrids have been recorded in nature. However, further investigation of An. longipalpis is still required with more field specimens.Koekemoer et al  reportedferences ,2. Howevrphology ,3. Koekey [et al  indicatet et al.  also repAn. funestus group and An. longipalpis type C.The application of the method described is expected to greatly improve the efficiency of large-scale analysis of field-collected samples of the The authors declare that they have no competing interests.KSC designed the study, developed the new RFLP assay and drafted the manuscript. MC and LLK assisted with analysis of the data and helped draft the manuscript.All authors have read and approved the final manuscript.


Solanum tuberosum L.) consists of a large number of tuber-bearing wild Solanum species under Solanum section Petota. One of the major taxonomic problems in section Petota is that the series classification (as put forward by Hawkes) is problematic and the boundaries of some series are unclear. In addition, the classification has received only partial cladistic support in all molecular studies carried out to date.The secondary genepool of our modern cultivated potato  were genotyped using over 200 AFLP markers. This resulted in the largest dataset ever constructed for Solanum section Petota. The data obtained are used to evaluate the 21 series hypothesis put forward by Hawkes and the 4 clade hypothesis of Spooner and co-workers.The aim of the present study is to describe the structure present in section We constructed a NJ tree for 4929 genotypes. For the other analyses, due to practical reasons, a condensed dataset was created consisting of one representative genotype from each available accession. We show a NJ jackknife and a MP jackknife tree. A large part of both trees consists of a polytomy. Some structure is still visible in both trees, supported by jackknife values above 69. We use these branches with >69 jackknife support in the NJ jackknife tree as a basis for informal species groups. The informal species groups recognized are: Mexican diploids, Acaulia, Iopetala, Longipedicellata, polyploid Conicibaccata, diploid Conicibaccata, Circaeifolia, diploid Piurana and tetraploid Piurana.Petota.Most of the series that Hawkes and his predecessors designated can not be accepted as natural groups, based on our study. Neither do we find proof for the 4 clades proposed by Spooner and co-workers. A few species groups have high support and their inner structure displays also supported subdivisions, while a large part of the species cannot be structured at all. We believe that the lack of structure is not due to any methodological problem but represents the real biological situation within section  Solanum tuberosum L.) consists of a large number of tuber-bearing wild Solanum species which grow in various habitats from the southern states of the USA to the most southern parts of Chile and Argentina. These wild species are important as a resource for valuable traits that can be used to improve the quality of the cultivars, including resistance against important diseases like Phytophthora infestans and potato cyst nematodes (Globodera spp.). Therefore it is no surprise that the wild relatives of the cultivated potato have since long drawn the attention of many plant breeders and botanists. To benefit most from the possibilities that the secondary genepool has to offer, it is necessary to have a good insight in the taxonomy. The classical treatments of potato taxonomy are from Correll [The secondary genepool of our modern cultivated potato  whose accessions did not cluster all together and 48 species whose accessions were mixed with accessions of other species. The latter was often the case with species that occur in South America, the borders of many of these species are not clearly recognizable from the NJ tree.S. nigrum, S. chaparense, S. sitiens, and S. fraxinifolium 2) North and Central American diploid series Polyadenia, Pinnatisecta, Bulbocastana and Morelliformia, 3) Circaeifolia and Piurana accessions, 4) Longipedicellata accessions, 5) Demissa and Conicibaccata accessions but without S. demissum and S. semidemissum, 6) S. verrucosum accessions, 7) Tuberosa from Bolivia, Argentina and Chile plus some accessions from other series such as Yungasensa, 8) accessions from cultivated Tuberosa species and wild Tuberosa from Peru, 9) accessions from Tuberosa and Megistacroloba, 10) accessions from S. acaule (and its subspecies), S. albicans, S. demissum, S x semidemissum and S. edinense.Above the species level, a few clusters of species groups can be distinguished in the large NJ tree . Roughly, the following groups can be found in the NJ tree of the large dataset: 1) an outgroup with Because of the size of the dataset, it proved impossible to analyze it with cladistic methods nor to analyze it for statistical support. A condensed dataset was created by choosing a representative genotype from all the available accessions (see methods section for exact details). This condensed dataset consisted of 916 genotypes.A single ratchet parsimony search consisting of 200 iterations yielded a Maximum Parsimony (MP) tree of 9669 steps. Furthermore, 20 individual independent ratchet searches each consisting of 50 iterations also yielded a MP tree of 9669 steps.Figure When comparing the NJ and the MP jackknife trees it is apparent that a large part of both trees consists of a polytomy. However, some structure is still visible in both trees, supported by jackknife values above 69. The following groups can be recognized in both the NJ jackknife tree and the MP jackknife tree:1) Mexican diploid species, with a jackknife support of 73 for the MP tree and 99 for the NJ tree; the substructure found within the Mexican and Northern American diploids is almost the same for both trees.Longipedicellata, with a jackknife support of 100 in both trees.2) A group of tetraploid Mexican/North and Central American species belonging to series S. acaule, S. demissum, and closely related species with a jackknife support of 100 in the MP tree and 99 in the NJ tree.3) A group consisting of accessions of Circaeifolia, with a jackknife support of 100 in both trees.4) A group consisting of the species belonging to series S. paucijugum, S. tuquerrense, and S. solisii, tetraploid species belonging to the series Piurana, with a jackknife support of 96 in the NJ tree and 92 in the MP tree.5) A small group of accessions belonging to There are also differences in group structure between the two trees. There are a number of groups that have good jackknife support in the NJ tree but are not supported in the MP jackknife tree:Demissa with a jackknife support of 79. In the MP tree only 2 species that are part of this group were found in one small clade: S. schenckii and S. hougasii.1) A group of hexaploid Mexican species belonging to series Conicibaccata has a jackknife support of 82 in the NJ jackknife tree. In the MP jackknife tree the same accessions are part of the polytomy. These clades represent the subgroups found within the Conicibaccata group in the NJ tree. Only one subgroup is not represented by a similar clade in the MP jackknife tree.2) A group of accessions from species belonging to series Piurana has a jackknife support of 69 in the NJ tree. In the MP tree, the jackknife support was low, so this group collapsed and 4 out of 5 supported subgroups found in the NJ jackknife tree are visible as supported separate small groups in the MP jackknife tree.3) A group of species belonging to series Conicibaccata, S. buesii, S. sandemannii and S. laxissimum with jackknife support of 92.4) A group consisting of accessions from diploid species of series S. medians, S. sandemanii, S. weberbauerii and an unknown species with a jackknife support of 85.5) A group which contains accessions of Lactuca species, sufficient phylogenetic signal was present and concluded that in practice the influence of possible limitations of AFLP, such as co-migration of nonhomologous fragments is limited. However, he stresses that the conclusion only applies to datasets with closely related species. Moreover, Kardolus [Solanum section Petota the AFLP technique is suitable up to the species level. The AFLP method has since then successfully been used in more studies on potato taxonomy [One of the arguments against the use of AFLP is the possible bias caused by homoplasy ,19,20. Ntaxonomy -24.S. demissum and S. acaule, the group of S. circaeifolium, the group of S. commersonii and the group of S. schenckii and S. hougasii. Then there are groups that are not supported in the MP jackknife tree  species do form a separate cluster.From the present results it is clear that there is no absolute relationship between EBNs and the groups found. In the group which contains Petota. Furthermore, there is a lack of support for the relationships between the different groups that were found in the NJ and MP trees. It is important to differentiate between these two phenomena because the causes underlying both cases could be different.With regard to the overall structure of the section as found in this study two main observations can be made. There seems to be a lack of supported structure, especially in the South American part of section The AFLP jackknife NJ tree and the jackknife MP tree in this study shows a lack of structure or rather, an unresolved structure for the part of the tree which contains South American species while the other part of the tree shows several well supported groups.Tuberosa different genotypes of the same species are not always grouped together and are scattered among genotypes from other species. He claims that the cause of this phenomenon is not the lack of resolution of AFLP, but the overclassification of a group of species, the so-called brevicaule-complex. The cpDNA RFLP studies of Spooner and Sytsma [Kardolus et al.  mentioned Sytsma , and Spod Sytsma  also shoSolanum section Petota and found high bootstrap values for the branch uniting all the South American species in three different types of dendrogram . However, the two subgroups within the South American clade that they distinghuished (variants C1 and C2) often show polytomies and resolution within the groups is mostly lacking.Volkov et al.  comparedLupinus. This would point at a rapid and recent diversification in the Andes. The authors also suggest that Lupinus is probably only one example of many plant radiations that followed the final uplift of the Andes. They assume that many of these plant radiations are yet unknown. It is possible that the factors underlying the Lupinus diversification are also responsible for the Solanum section Petota diversification. According to Hughes and Eastwood [Outside the field of potato taxonomy, researchers have reported similar patterns. Hughes and Eastwood  report aEastwood  these faS. etuberosum, S. palustre and S. fernandezianum which connects to the main branch of the NJ jackknife and MP jackknife tree with respectively 100 or 98 support value, none of the branches connecting two or more groups have jackknife support of 69 or higher. That is the reason why in the schematized jackknife NJ and jackknife MP trees these branches collapse in a polytomy. Contrastingly, the branches of the groups that can be recognised within the polytomy do have jackknife support, although not all species can be put in groups as discussed previously.Except for the outgroup consisting of Petota taxonomy by Kardolus et al. [Petota. Bootstrap support above 70 were given for a NJ tree branch connecting the outgroup of S. etuberosum and S. brevidens, for a branch connecting the outgroups, and for the Mexican diploids and S. circaeifolium and S. circaeifolium subspecies quimense with the other part of the tree. In the cpDNA RFLP studies on the South American part of section Petota [S. cardiophyllum and S. bulbocastanum) is connected to the other clades with a bootstrap value of 87, and Clade 3 (mainly accessions belonging to series Piurana) and Clade 4 (the rest of section Petota) are connected to each other with a branch with 96 bootstrap support.In the first study on the use of AFLP in s et al. , it provn Petota  only a fPetota in general. This indicates that our and previous results represent the real biological situation in Solanum section Petota. Since the phylogenetic signal is clearly present in our data as shown in the well-supported groups in the present study, the lack of structure in parts of the tree is not caused by the lack of phylogenetic signal in AFLP markers.We can conclude from these previous results that it is indeed difficult to find good support for the backbone structure of section Petota in 21 series. Although a few of the series seem to form natural groups, the majority of the series as proposed by Hawkes [Petota.As outlined in this paper and in other earlier studies, there are no results that support the classification of section Petota in informal species groups, following the approach of Spooner et al. [We propose to divide section r et al.  who consr et al.  and Knapr et al. ,30 who ar et al.  if applir et al. .In total, the NJ jackknife tree can be partitioned into 10 species groups. It would be possible to construct more species groups based on the structure shown in the various trees made in the present study, but these groups would then not be supported by bootstrap or jackknife supports.Although a closed classification following the rules of the Botanical Code is desirable, it seems in this case difficult to apply. In the present study, many species cannot be accommodated in groups. These species do not automatically form a group themselves, but are intentionally left unclassified.We suggest recognizing the following informal species groups as shown in the NJ jackknife tree Figure :This group contains the species groups of Spooner et al. : PinnatiS. semidemissum, S. demissum and S. x edinense. The other group shows a jackknife support of 98 and contains S. juzepczukii, S. albicans and the three subspecies S. acaule subsp. acaule, S. acaule subsp. aemulans, S. acaule subsp. punae.In our study this group contains 2 supported subgroups, one branch with jackknife support of 96 containing the species S. schenckii, S. hougasii, that form a strongly supported cluster together (jackknife support 100) and a cluster containing the species S. iopetalum, S. brachycarpum, S. guerreroense (jackknife support 90). All species were formerly designated by Hawkes [Demissa which also included the species S. demissum and closely related species. The species in our group are the same as in the species group Iopetala designated by Spooner et al. [S. brachycarpum as a synonym of S. iopetalum.This group contains the species y Hawkes  to serieLongipedicellata. The species included in this group are S. fendleri including S. fendleri subsp. arizonicum, S. stoloniferum, S. hjertingii,. S. papita, S. polytrichon, S. leptosepalum, and S. matehualae. The species S. leptosepalum, S. fendleri, S. papita, and S. polytrichon have been reduced as synonyms of S. stoloniferum [S. matehualae is reduced as synonym of S. hjertingii [As the name does suggest, this group contains species that were formerly placed by Hawkes  in the soniferum . The speertingii .Conicibaccata. According to the present study the group consists of S. flahaultii, S. moscopanum, S. orocense, S. sucubunense, S. tundalomense, S. oxycarpum, S. longiconicum, S. garcia-barrigae, S. otites, S. oxycarpum, S. agrimonifolium, S. moscopanum, S. subspanduratum, S. paramoense, and S. colombianum.This group contains species placed there by Spooner et al. , complemConicibaccata can be put in the species group Conicibaccata there are a few species that form a separate group. This group consists of the diploid species S. buesii, S. sandemanii, and S. laxissimum.Although most of the series Piurana as the contents of the group are roughly similar: S. piurae, S. acroglossum, S. blanco-galdosii, S. irosinum, S. chomatophilum, and S. paucissectum from series Piurana and S. chiquidenum from series Tuberosa.This species group was not designated by Spooner et al. . The namPiurana series [S. paucijugum, S. tuquerrense, and S. solisii.The situation as described before for the Conicibaccata group also applies partly for the Piurana group. There are a few species from the formerly designated a series  that forS. circaeifolium, S. soestii, S. capsicumbaccatum and S. circaeifolium subsp. quimense. The contents is conform Hawkes' series Circaeifolia.This group consists of S. macropilosum and S. verrucosum. The species S. macropilosum was reduced to a synonym of S. verrucosum by Spooner et al. [This group contains only 2 species; r et al. .Solanum section Petota accessions ever analysed simultaneously. All other previous studies used datasets that included less variation and fewer species. Because of the thorough sampling, it is possible to propose species groups without too many reservations. A number of species groups coincide with certain series recognized by Hawkes [Solanum section Petota is highly unbalanced. A few species groups have high support and their inner structure displays also supported subdivisions, while a large part of the species cannot be structured and they seem to be all equally related to each other and to the supported groups.As far as we know, this paper treats the largest collection of y Hawkes . HoweverSolanum section Petota cannot be structured or subdivided. We even doubt that it would be possible to find more resolution with other methods or more markers, and we consider it likely that the polytomy is indicative of the real situation in section Petota. A relatively fast spread of tuber-bearing Solanum species over South America, due to the geographic conditions in the Andes [It might be difficult to accept that a part of genus he Andes , combineSolanum clones was grown in vitro for at least 6 weeks on MS medium supplemented with 20% sucrose [In total 951 accessions representing 196 different taxa, species, 15 subspecies and 17 hybrids were sampled. We tried to include as many species as possible from various gene banks. In principle, at least 5 accessions from each available species and 5 individual plants per species  were included. Seeds were surface-sterilized and sown in vitro at 25°C. The collection of individual  sucrose  at 18°C. sucrose .S. spec.. The accessions which labels were changed are indicated in Additional file Additional file EcoRI/MseI AFLP primer combinations: E32/M49 and E35/M48. The protocol of Vos et al. [The samples were fingerprinted with two s et al.  was usedThe dataset in this study originally contained 4929 genotypes. This large dataset was analyzed with NJ and UPGMA. Because of the size of the dataset, it proved impossible to analyze it with cladistic methods nor to analyze it for statistical support, even using the SARA supercomputer (see below). It was sheer impossible for a personal computer to do any further analyses apart from the NJ and UPGMA, and for the SARA computer cluster it would have taken many months/years of computing time.For further analysis a condensed dataset was created by carefully choosing a representative genotype from all the available accessions. This condensed dataset consisted of 916 genotypes. The condensed dataset was used in both phenetic and cladistic analyses and in the resampling methods.S. sitiens, S. nigrum, S. chaparense, S. lycopersicoides, S. canense, S. fraxinifolium). The outgroup species S. etuberosum, S. palustre and S. fernandezianum were retained in the dataset.Besides choosing only one genotype per accession to represent the accession in the condensed dataset, other adjustments were made to create this dataset. All the 22 known interspecific hybrid accessions were removed, 23 other accessions were completely removed because of the extreme heterogeneity of the accession (possibly resulting from a mixture of species) in both the NJ and the UPGMA trees. Species labels of 49 accessions were changed based on their position in the NJ and/or UPGMA tree (not shown) and visual inspection of the plants in the experimental field or greenhouse in 2005 and 2006. In total 11 outgroup accessions were removed because preliminary AFLP results showed these outgroups to be too distant (Both the phenetic and the cladistic analyses were conducted using PAUP 4.0 Altivec  on the TThe cladistic analysis heuristic searches were done by using PRAP, Parsimony Ratchet Analyses using PAUP, a program that writes commands for PAUP. The commands in PRAP describe how PAUP should carry out parsimony ratchet searches . By usinFor the MP jackknife analysis, we followed the conclusions drawn by Muller  that usiMJ carried out the phenetic and phylogenetic analyses and drafted the manuscript, RVB and BV participated in coordination and design of the study and participated in drafting the manuscript, RH chose the plant material and participated in coordination and design of the study, VV and MV coordinated and carried out the growing of the material, MS and RM carried out the AFLP analyses. All authors read and approved of the final manuscript.Symbols used in additional file 1. # recorded hybrid, removed in 916 dataset. $ complete accession removed in the 916 dataset because of conflicting positions in NJ tree. & removed outgroups in 916 dataset: S. lycopersicoides, S. nigrum, S. chaparense, S. sitiens, S. canense, S. fraxinifolium. * the label of this accession was changed in the 916 dataset after checking the position in the large NJ tree and checking morphology in the greenhouse/field.  the number in parentheses indicates the number of accessions used for the 916 analysis in case of removal or change of accessions. Abbreviations for Genebank source codes: CPC: Commonwealth Potato Collection, UK. CGN: Centre for Genetic Resources, the Netherlands. cgn: cgn receipt number, Centre for Genetic Resources, the Netherlands. PI: Plant Introduction number, USA. GLKS: Gross Lusewitz, Germany.List of material used in the AFLP analyses. List of material used in the AFLP analyses. Click here for file


Anaplasma phagocytophilum, Borrelia burgdorferi, Dipylidium caninum, Leishmania infantum, Dirofilaria immitis, and Dirofilaria repens) are of zoonotic concern. Recent studies have highlighted the potential of fleas as vectors of pathogens of zoonotic relevance  in this country. While some arthropod vectors  are present in certain Italian regions throughout the year, others  are most active during the summer season. Accordingly, control strategies, such as those relying on the systematic use of acaricides and insecticides, should be planned on the basis of the ecology of both vectors and pathogens in different geographical areas in order to improve their effectiveness in reducing the risk of infection by vector-borne pathogens. This article reviews the current situation and perspectives of canine and feline vector-borne diseases in Italy.In Italy, dogs and cats are at risk of becoming infected by different vector-borne pathogens, including protozoa, bacteria, and helminths. Ticks, fleas, phlebotomine sand flies, and mosquitoes are recognized vectors of pathogens affecting cats and dogs, some of which (e.g.,  Canine and feline vector-borne diseases (VBDs) are caused by a wide range of pathogens, including viruses, bacteria, protozoa, and helminths, which are transmitted by a variety of vectors, such as ticks, fleas, mosquitoes, and phlebotomine sand flies. Some VBDs might be life-threatening in cats and dogs, they might develop after long incubation period making their diagnosis challenging, particularly because their clinical signs are not pathognomonic ,2. In adIn Italy, as well as in many other European countries, the aforementioned key information on parasitic arthropods and the pathogens they transmit is scant, anecdotic, and often outdated . This isThe present article reviews the current situation and perspectives of canine and feline VBDs in Italy. Additionally, the risk of transmission of vector-borne pathogens through different seasons is discussed in order to afford the implementation of effective control programs.Italy is located in southern Europe and comprises the long, boot-shaped Italian Peninsula, the land between the peninsula and the Alps, and two main islands (Sicily and Sardinia). The country has a territory of 301,230 sq km, of which 294,020 sq km is land. It is geographically differentiated into three main areas which include administrative regions, namely northern , central , and southern Italy  including Sicily and Sardinia islands  dogs and cats are often present in urban and rural areas representing a public health concern in Italy . Stray dCats and dogs living in public shelters in Italy may be at high risk of acquiring vector-borne pathogens, mainly because they are often untreated against ectoparasites, thus, representing an easy feeding source for them. In addition, the general conditions of these animals  may contribute to susceptibility to some VBDs. Likewise, when infected, free-roaming cats and dogs are often neither monitored nor treated against vector-borne pathogens.vice versa poses a risk for the introduction and dissemination of exotic pathogens if competent vectors are present. In a recent study, it was found that 62% of dogs infected by Babesia canis had a history of travel to East European countries .D. immitis were thought to be more active during the summer months, mainly in humid climate areas of northern Italy and, at a lesser extent, in the remaining part of the Peninsula [A. albopictus, a known vector of D. immitis and D. repens [A. albopictus has adapted to the relatively low winter temperatures of Italy, rapidly increasing its populations through the country, developing many generations over the year in central and southern regions and overwintering as eggs in colder northern regions [A. albopictus has a diurnal activity pattern [Dirofilaria spp. in areas where both A. albopictus and other nocturnal mosquitoes  occur in sympatry [For a long time, mosquito vectors of eninsula . However. repens ,42, duri. repens . In addi regions . In cont pattern . This posympatry .P. perniciosus (the most important vector of L. infantum in the Mediterranean area) is restricted to June and early October [The occurrence and spread of canine leishmaniosis in a given area depends on several factors, including vector abundance and biting rates. In Italy, the activity of phlebotomine sand flies is seasonal and restricted to the summer months -47. Inde October . The act October ,47.P. variegata has been studied in an area in southern Italy where T. callipaeda is highly endemic [P. variegata has a seasonal pattern from May to October, being more active during July and August, at 20-25°C and 50-75% of relative humidity [The ecology and distribution of  endemic . It has humidity .E. canis infection in dogs estimated by serological surveys varied from 14.9% in southern Italy [E. canis infection among 601 Italian dogs estimated by real-time PCR was lower than that recorded by serology, indicating that the infection is most prevalent in southern (9.7%) than in central (8%) and northern (2.9%) Italy [E. canis infection varies among foci according to local factors . Anaplasma platys infection has been molecularly detected in kennelled dogs in central (23%) [E. canis and A. platys may also occur as recorded in a dog population from southern Italy where 44.4% of individuals infected by A. platys were co-infected with E. canis [Rhipicephalus sanguineus, a proven vector of E. canis, has been suspected to act a vector of A. platys in Italy [E. canis and A. platys adds weight to this hypothesis. Similarly, high prevalence rates of anti-R. conorii antibodies (up to 74%) have been detected in dogs [R. conorii (56%) and A. phagocytophilum (3.7%) in dogs [Bartonella spp. DNA [Bartonella sp. (strain HMD) has been detected in five dogs, one of which was co-infected with B. vinsonii berkhoffii (genotypes II and III) [Bartonella species, some of which being of human health concern .It is difficult to assess the actual prevalence of infection by different vector-borne pathogens in cats and dogs in Italy due to the limited amount of data as well as to the difficulties in comparing information from studies using different diagnostic tools. Indeed, the prevalence of infection by vector-borne pathogens can also vary according to geographical region. For instance, the prevalence of rn Italy  to 46.7%rn Italy . Convers%) Italy . Indeed,al (23%)  and soutal (23%)  as well al (23%) . In addiE. canis . Rhipicein Italy  and the  in dogs . A study in dogs . In a respp. DNA . Moreoveand III) . These dBabesia protozoa infecting dogs in Italy [Babesia antibodies in dogs with a decreasing trend from central to northern areas [Babesia spp. infection [Babesia gibsoni in Italy has recently been supported by molecular data [Babesia spp. in blood samples from dogs with clinical signs compatible with VBDs have shown that B. canis is mainly detected in northern Italy (29.1%) whereas B. vogeli was detected mainly in central and southern Italy (16.3%) [There are limited data on in Italy -54. In arn areas . In the nfection . The occlar data . Again, D. repens has been considered for a long time to be mainly diffused in southern regions, D. immitis is endemic in northern regions with prevalence rates ranging from 22 to 80% [D. immitis showed a relevant prevalence increase in endemic areas [D. immitis has also become endemic in central regions such as Tuscany and Umbria [A. reconditum (16.5%) followed by D. repens (1.4%) and D. immitis (0.5%) in Campania [D. immitis and D. repens has been also recorded in Apulia and Calabria with prevalence rates up to 1.6% [D. repens in northern Italy [D. immitis recently detected in southern regions [Dirofilaria spp. throughout the whole country. Until now, D. immitis infection in cats has been diagnosed mostly in northern Italy where the prevalence is about 18% in pet cats [While canine dirofilariosis by 2 to 80% ,56 in doic areas . Furtherd Umbria . A recenCampania . The occ to 1.6% . The sprrn Italy  and the  regions  indicatepet cats .T. callipaeda has also been found among different wildlife species, which have been implicated as its reservoirs [The highest prevalence of canine thelaziosis has been reported in some areas of southern Italy (Basilicata region), reaching up to 60% in certain municipalities . In thisservoirs .For long time, stable endemic foci of canine leishmaniosis have been reported in southern and central Italy, with seroprevalence rates reaching up to 53.1% in some foci . In a stA. phagocytophilum, Bartonella henselae, B. vinsonii berkhoffii, B. burgdorferi, Coxiella burnetii, R. conorii, and R. felis), nematodes , tapeworms (D. caninum), and L. infantum. In most of the cases, cats and dogs are unlikely to represent important reservoir hosts and their role in the transmission of these pathogens to humans is probably minor. On the other hand, cats and dogs may play a key role in the zoonotic cycle of transmission of some pathogens, such as B. henselae and L. infantum, respectively.Many vector-borne pathogens infecting cats and dogs in Italy may also be a threat to human health. These include many bacteria  of public health concern in Italy [Dogs are primary hosts of vector-borne pathogens that may occasionally affect humans in Italy, including . repens ,66, and llipaeda . Most iminfantum ,68,69, winfantum . Althougin Italy ,66. In p. repens  and dogs. repens . Similarountries . Althougin Italy , their rB. henselae and Bartonella clarridgeiae in Italy [L. infantum has long been discussed [L. infantum infection to P. perniciosus under laboratory conditions [R. felis (the causative agent of flea-borne spotted fever in humans) in fleas (C. felis and C. canis) collected from cats [Cats have been found infected by emerging human pathogens, such as in Italy , and migiscussed  and a renditions . Recent rom cats ,27. HoweConsidering the zoonotic relevance of many vector-borne pathogens infecting cats and dogs in Italy, the control of VBDs they cause should not be only of veterinary concern but also a public health priority.The control of VBDs requires a holistic approach, considering the distribution, and the ecology of the vectors and of pathogens they may transmit and the infection progression in infected animals. The current strategies for the control of arthropod vectors in dogs and cats have recently been reviewed elsewhere ,74. The The prevention of VBDs can be achieved by means of systematic application of acaricides and/or insecticides, prophylactic administration of drugs, and vaccination. However, as discussed previously in this review, the occurrence and the risk of acquiring VBDs in Italy can vary according to the geographical and seasonal distribution of their respective vectors. Thus, the success of any prevention strategy depends on the appropriate use of commercially available tools, which should not only consider the manufacturer's instructions but also the available information on the vector ecology in a given area or region.D. immitis and D. repens has long been recommended for dogs in highly endemic areas of northern regions [E. canis [L. infantum in dogs [B. burgdorferi sensu stricto, but not against the other species. The decision to vaccinate against Lyme borreliosis should be made based on a risk assessment of the individual dog that includes information about where the dog lives and how often it frequents a tick-infested area. As far as cats, no studies are available in the literature about the prevention of VBDs.In Italy, the chemoprophylatic use of ivermectin against third- and fourth-stage larvae of  regions . In soutE. canis  and L. i in dogs ,76. In IR. felis) almost no information is available.Undoubtedly, the general picture of the VBDs affecting dogs and cats in Italy is complex mainly due to the limited amount of published data on ecology and distribution of arthropod vectors and the pathogens they transmit in northern, central and southern areas of the country. Gaps in our knowledge are even worse for vector-borne pathogens infecting cats; for some of which  [R. sanguineus in southern Italysuggests that unrecognized vector-borne pathogens may exist, which could complicate the diagnosis and management of other endemic VBDs in dogs and cats. Furthermore, considering the importance of R. sanguineus as vectors of R. conorii and the medical relevance of R. conorii infection in Italy [Among arthropods parasitizing dogs and cats in Italy, ticks are the most important vectors of pathogens. Indeed, ticks are proven vectors of a large number of pathogens, present in a wide geographical distribution and, most importantly, they are active during the whole year. In particular, due to its high degree of adaptability to different microenvironments and its capability to occasionally feed on hosts other than dogs, d humans ,11. Furtnt years ,77. For ngestion  and a rern Italy . Furtherain HMD)  in salivin Italy ,81, furtAs a priority, it is necessary to establish effective national surveillance systems based on a regional level that, using standardized diagnostic procedures and protocols, could provide a clear picture on vector-borne pathogens circulating among dogs and cats (both pet and free-roaming ones) in different Italian regions. Data on the distribution of arthropod vectors  and the pathogens they transmit would not only allow the assessment of the risk of the introduction of exotic pathogens into non-endemic areas but, more practically, could provide information to veterinarians and pet owners on timing and protocols that should be chosen for preventing VBDs in different Italian regions. In this regard, it is crucial to translate that data from research into changed practices by veterinarians and to ensure owners' compliance.An affordable and reliable control of canine and feline VBDs should also be part of a strategy for the management of zoonotic infections in order to reduce the risk of pathogen circulation between pet animals and humans. However, these actions cannot be disconnected from political initiatives to reduce the number of free-roaming dogs and cats in Italy and to improve vector control programs in public kennels. Finally, it is crucial to implement a permanent surveillance system to prevent the entry and exit of exotic vector-borne pathogens through dogs and cats travelling with their owners to and from Italy.The authors declare that they have no competing interests.DO and FD-T contributed equally to conceiving and writing this article.


Most research pertaining to childhood obesity has assessed the effectiveness of preventative interventions, while relatively little has been done to advance knowledge in the treatment of obesity. Thus, a 4-week family- and group-based intervention utilizing group dynamics strategies designed to increase cohesion was implemented to influence the lifestyles and physical activity levels of obese children.This paper provides an overview of the rationale for and implementation of the intervention for obese children and their families. Objectives of the intervention included the modification of health behaviors and cohesion levels through the use of group dynamics strategies. To date, a total of 15 children  and their families have completed the intervention (during the month of August 2008). Physiological and psychological outcomes were assessed throughout the 4-week intervention and at 3-, 6-, and 12-month follow-up periods.It is believed that the information provided will help researchers and health professionals develop similar obesity treatment interventions through the use of evidence-based group dynamics strategies. There is also a need for continued research in this area, and it is our hope that the Children's Health and Activity Modification Program (C.H.A.M.P.) will provide a strong base from which others may build. Canadian children are becoming progressively overweight and obese. In 2004, 26% of children and adolescents aged 2 to 17 were either obese or overweight . A seconprevention of obesity in youth ) worked together to complete a "C.H.A.M.P. Family Action Plan" that included the following prompts: (a) These are the reasons my family is involved in C.H.A.M.P.; (b) Our family's top 3 goals for the next 4 weeks are...; and (c) State the actions your family will take over the next 4 weeks to reach your goals. Insofar as the last element is concerned , families were asked to outline: the During the family education sessions and the follow-up booster sessions, weekly messages, resources, and handouts regarding healthy living, ideas for physical activity, and motivation were provided to families (parents were provided with C.H.A.M.P. binders in which to store this information). These sessions also allowed parents and guardians to openly share and discuss their successes and difficulties in the home environment, and to ask questions to other group members or health care professionals. To further enhance communication among children, family members, and camp/research staff during the 4-week intervention, a weekly newsletter containing pictures, information about the previous week, health-related information, and upcoming events was provided to families. Each newsletter also contained a page entitled "The Fabulous You", which was dedicated to recognizing an outstanding accomplishment for each child during the previous week. As mentioned previously, families also received bi-monthly newsletters containing health- and program-related information following completion of the formal 4-week intervention.One of the primary purposes of both the family-based education sessions and the follow-up booster sessions was to provide families with an opportunity to meet, communicate, and interact with other individuals in a similar situation to themselves. Thus, all aspects of the family-based sessions centred around communication, feedback (where necessary), and social support. For example, parents were asked to share their contact information with one other C.H.A.M.P. family to enhance accountability for their family goals, and to increase communication, interaction, and support among families. Parents also participated in focus groups during which they were encouraged to openly share their perceptions of and experiences with the program. Finally, parents provided weekly feedback  to researchers about their involvement in the program.Open lines of communication were also maintained among counselors, program staff, and families throughout and following the 4-week program. When necessary, parents were provided with verbal or written updates regarding their child, and families were also given contact information for two counselors and several members of the research team. Following program involvement, families were contacted by program staff on a regular basis as a means of communication and support, and to relay friendly reminders for program-related events, sessions, and research assessments.For all family-based sessions, barriers to physical activity and healthy eating were identified and group discussions were implemented to create solutions for overcoming such challenges.To increase physical activity, group support, and family commitment, all families were provided with a one-month membership  at the YMCA in London, Ontario. In addition, one of the follow-up booster sessions consisted of a family-based aerobics/strength training session for children, siblings, and guardians. Finally, families were provided with coupons and information for family-friendly activities  that could be utilized following involvement in the 4-week program.z), waist circumference, lean body mass, body fat percentage , and specific fitness indices . Fasting blood sample measurements yielded values for total cholesterol, high density lipoprotein cholesterol (HDL-c), low density lipoprotein cholesterol (LDL-c), triglycerides, insulin, serum glucose, and HOMA-IR. Additionally, vessel wall imaging (VWI) was conducted on all children to monitor plaque formation, vessel elasticity, and capillary function, thus providing an estimate of cardiovascular health. As mentioned previously, semi-structured focus groups were also conducted with children and parents to gain qualitative information pertaining to participants' experiences with and the perceived impact of the program. Finally, self-report measures included children's health-related quality of life (measured via the PedsQL 4.0) [A mixed methods  approach was taken in the collection and evaluation of data generated from the intervention and follow-up assessments. Several outcomes were assessed at various time points including standardized body mass index (BMI-run test . ObjectisQL 4.0) -40, the sQL 4.0) , questiosQL 4.0) , task ansQL 4.0) , and perRate each of your C.H.A.M.P. camp-mates on how much of a friend they are to you". Children were shown pictures of all C.H.A.M.P. participants, and asked to rate each child on a scale from 1 ("very good friend") to 5 . The remaining 15 questions were answered on a five-point response scale , with a corresponding graphic scale containing faces with a range of emotional expressions  adapted from Wong and Baker [the total C.H.A.M.P. population , the C.H.A.M.P. teams , and the C.H.A.M.P. counselor groups . As can be seen in Table S1 (additional file Cohesion was measured using an 18 item questionnaire developed specifically for the present research and modified from the Sports Cohesiveness Questionnaire . The firnd Baker . These qIn summary, this article has provided a rationale for the creation of C.H.A.M.P., a background in the area of childhood obesity, and an overview of the group dynamics strategies used to facilitate cohesion among children and family members. All of the information needed to create and implement a similar group-based program has been discussed: study design, components of the intervention, recruitment, and measurement procedures. Although data analyses from Year 1 are not complete, preliminary results appear promising and anecdotal reports from children, guardians, and counselors have been positive. Additionally, the first year of this pilot project has provided valuable information regarding what aspects of the intervention appear to be most and least effective. The results of the analyses will help further the knowledge in family- and group-based interventions aimed at the treatment of overweight and obese children. Results of the intervention will be available in late 2009, and will be disseminated to relevant community and government organizations, and published in academic journals. In the meantime, it is hoped that the issues discussed provide guidance to those undertaking similar trials with children. Ultimately, it is our goal to utilize the results of the 2-year pilot project to offer C.H.A.M.P. on an annual basis with a larger number of children and families across a broader range of ages, settings, and communities.The authors declare that they have no competing interests.SB, JI, RP, HP, and KS were all involved in the development of the program. They provided support and expertise related to the design and implementation of the study. LM, SB, AC, and SS participated in the writing of the manuscript, while JI, RP, HP, and KS provided revisions and comments, and approved the final version. All authors read and approved the final version of the manuscript.The pre-publication history for this paper can be accessed here:Table S1.Click here for file


Sir,It seems that the authors were completely oblivious of the landmark paper which was published by Dr. A. D. Dias on the "SEPA" flap.st paper was published in the British Journal of Plastic Surgery 1984;37:256-61 as "The Superficial External Pudendal Artery (SEPA) axial pattern flap". In this paper, the anatomy of the flap was studied in 12 cadavers and the flap has was clinically in six patients. A bilateral SEPA flap was also described. Moreover, the anatomical position of the flap makes it possible to immobilize the hand in a much more comfortable and natural position than that assumed in a groin flap. As the flap can be rotated to almost 180 degrees on either side, it is very suitable for defects in the genital region. Thus, it can be used for skin cover in the second stage repair of hypospadius, or a urinary fistula, or in difficult repairs in a hypospadic cripple. It can also add bulk to a hypoplastic penis and its use in epispadias repair is very convenient. The flap is also useful for degloving injuries of the penis and scrotum, in reconstruction of an amputated penis and in the correction of certain contractures in the groin.The 1A second paper: "The Anatomical basis of the SEPA flap" by U. A. Patil, A. D. Dias and R. L. Thatte was published in the British Journal of Plastic Surgery 1987;40:342-7.A third paper: "The use of the SEPA flap in the repair of defects in the hands and fingers" by A. D. Dias, R. L. Thatte, U. A. Patil, L. D. Dhami, S. Prasad was also published in the British Journal of Plastic Surgery 1987;40:348-59.I hope that I have placed the subject in the proper perspective.


To the Editor: Tickborne rickettsioses are caused by obligate intracellular gram-negative bacteria that belong to the spotted fever group of the genus Rickettsia. These zoonoses share characteristic clinical features, including fever, headache, rash, and sometimes eschar formation at the site of the bite  of rickettsia, as described (ompA gene (R. montanensis DNA]) were included in each test. The sequences of PCR products were obtained and compared with those available in GenBank  similarity with R. conorii (GenBank accession no. AY346453); those of a 340-bp segment of gltA showed 100% similarity with that of R. conorii (AE008677). The sequences of a 517-bp segment of ompA and a 341-bp segment of gltA amplified from A. lepidum showed 100% similarity to the corresponding sequences of R. africae .One specimen of R. conorii, the agent of Mediterranean spotted fever, in Uganda. Although the main vector of this rickettsia is Rh. sanguineus, the few ticks of this species we tested were negative (R. conorii in Ha. punctaleachi, although it has been detected in the closely related Ha. leachi in Zimbabwe (Ha. punctaleachi prefers warm and humid conditions but can exist wherever rodent hosts for its immature stages and canine hosts for its adult stages are present (R. conorii in Ha. punctaleachi does not mean that this tick is an efficient vector (To our knowledge, this is the first detection of R. africae, the agent of African tick bite fever, in Djibouti. R. africae was also detected in 1 A. lepidum collected in Sudan (A. variegatum and A. hebraeum with high infection rates throughout sub-Saharan Africa (A. lepidum, which coexists with A. variegatum in limited locations, is chiefly a cattle parasite. It will also attach to smaller domestic animals and a few wild herbivores, but it attacks humans less frequently than A. variegatum or A. hebraeum. A. lepidum occurs in a variety of climatic regions but most commonly inhabits semiarid regions in eastern Africa. The cattle in our study had been imported from Ethiopia, and the ticks may have infested these animals before their arrival in Djibouti. Indeed, in 1973 Burgdorfer obtained an isolate from A. variegatum in Ethiopia, which was thereafter shown to be indistinguishable from the rickettsia described as R. africae (R. africae in Djibouti and that it could affect their patients, both local and international, including French and American soldiers based in this country (This is also, to our knowledge, the first detection of R. conorii and R. africae in Uganda and Djibouti, respectively. However, healthcare workers who treat persons who live in or have traveled to these countries should be alert for spotted fever group rickettsial infections in their patients (Because we did not do systematic sampling, our results cannot address the prevalence and distribution of