Leishmania that infected phlebotomines in Caxias, Maranhão, Brazil, an area that is highly endemic for leishmaniasis. Sandflies were captured in light traps located in the peridomiciliary environments of randomly selected houses in urban and rural settings between 1800 and 0600 hours on new moon days between March 2013 and February 2015. DNA extracts from 982 engorged female sandflies were submitted to fragment length polymorphism analysis to identify infecting species of Leishmania, and blood sources were identified for 778 of these specimens. Infection by Leishmania infantum was detected in Lutzomyia longipalpis, Lu. whitmani and Lu. termitophila; L. infantum/L. braziliensis in Lu. longipalpis, Lu. whitmani and Lu. trinidadensis; L. shawi in Lu. longipalpis; L. mexicana in Lu. longipalpis; L. braziliensis in Lu. longipalpis and Lu. whitmani; L. guyanensis in Lu. longipalpis and Lu. termitophila; L. amazonensis in Lu. longipalpis and L. lainsoni or L. naiffi in Lu. longipalpis, while Lu. longipalpis and Lu. trinidadensis were infected with unidentified Leishmania sp. Blood sources were identified in 573 individual phlebotomines and the preferred hosts were, in decreasing order, chicken, dog, rodent and human with lower preferences for pig, horse, opossum and cattle. Lu. longipalpis and Lu. whitmani performed mixed feeding on man, dog and rodent, while Lu. longipalpis was the most opportunistic species, feeding on the blood of all hosts surveyed, but preferably on dog/chicken, dog/rodent and rodent/chicken. Our findings reveal the concomitant circulation of Leishmania species that cause visceral leishmaniasis and tegumentary leishmaniasis in the study area, and explain the occurrence of autochthonous human cases of both clinical forms of leishmaniasis in Caxias, Maranhão. The results support our hypothesis that, in the municipality of Caxias, transmission of Leishmania occurs in close proximity to humans.The aims of the study were to determine the blood feeding preferences of sandflies and to identify species of  Leishmania (Kinetoplastida: Trypanosomatidae) and transmitted by hematophagous sandflies of the subfamily Phlebotominae (Diptera: Psychodidae)[Leishmania (Leishmania) infantum, and American tegumentary leishmaniasis (ATL), which manifests in cutaneous, mucocutaneous and diffuse forms [The leishmaniases are a group of neglected tropical diseases caused by protozoa of the genus chodidae). The dischodidae). The epichodidae). In the se forms ,4.Lutzomyia longipalpis, an anthropophilic phlebotomine that is widely distributed throughout the country, and Lu. cruzi, which is prevalent in some municipalities in the central-west region [L. infantum are domestic dogs and other wild canids [In Brazil, the principal vectors of the AVL pathogen are t region ,6. The md canids .Leishmania species responsible for tegumentary forms of the disease in humans. Six of these species belong to the subgenus Viannia: L. (Viannia) braziliensis, L. (V.) guyanensis, L.(V.) lainsoni, L.(V.) naiffi, L.(V.) shawi and L.(V.) lindenbergi, while L.(Leishmania) amazonensis belongs to the subgenus Leishmania [Lu. intermedia, Lu. neivai, Lu. whitmani, Lu. umbratilis, Lu. flaviscutellata, Lu. antunesi, Lu. migonei, Lu. fischeri, Lu. pessoai, Lu. wellcomei, Lu. complexa, Lu. ayrozai, Lu. paraensis, Lu. amazonensis, Lu. hirsuta hirsuta, Lu. ubiquitalis, Lu. gomezi and Lu. tuberculata [Regarding ATL, Brazil is the country with the greatest variety ishmania ,7. The serculata .Leishmania species within Brazil is not yet fully know, the geographical spread of leishmaniasis has expanded over recent years in several states, including the northeastern state of Maranhão where AVL and ATL are endemic and widespread [Leishmania have already been identified in Maranhão as, for example, Lu. whitmani infected with Leishmania sp. [Lu. longipalpis infected with L. infantum [Lu. longipalpis naturally infected with the AVL pathogen was 0.76% in the municipality of Caxias, Maranhão, where both AVL and ATL are prevalent. Despite this low infection rate, AVL endemicity in the area was sustained as demonstrated by the notification of 45 human cases of the disease during the year of the study [Leishmania responsible were not identified.Although the distribution of despread . Phlebotania sp.  and Lu. infantum ,12. Accohe study . In addiLeishmania sp.  associated with these sandflies and to determine the blood food sources of the infected insects. Molecular techniques afford high sensitivity in detecting Leishmania sp. and identifying the blood source of infected phlebotomines. Such information facilitates the understanding of vector competence, transmission dynamics and the ecoepidemiology of leishmaniasis, thereby increasing our knowledge of how species interact with their natural habitat and assisting in the guidance of disease control and surveillance [Considering that 91 phlebotomine species are distributed within the different ecosystems of Maranhão [eillance –17.Leishmania that infect phlebotomines and on the blood food sources of these vectors in the cerrado area of Caxias, Maranhão. Our results support the hypothesis that, in the municipality of Caxias, the transmission of Leishmania occurs in environs close to human dwellings. We have verified this hypothesis by carrying out an ecological study in which we have determined the frequency of phlebotomine species and identified the host preferences of the sandflies in urban and rural settings of Caxias.Few studies have focused on the identification of species of Didelphis albiventris) was authorized by the Ethics Committee on the Use of Animals in Research of the Universidade Federal de Minas Gerais (license no. 37950–1), while the collection of phlebotomines was approved by the Instituto Chico Mendes de Conservação da Biodiversidade (authorization no. 46319–1).Details of the project were submitted to and approved by the Ethics Committee of the Universidade Estadual do Maranhão (protocol no. 909.095). The collection of blood samples from the marsupial  located in the eastern mesoregion of Maranhão State. The municipality comprises an area of 5,150,647 kmabitants . The warabitants , and then = 10) and in the rural district of Bom Jardim (n = 10). Sampling was performed near the shelters of domestic animals, when present, with the aid of Centers for Disease Control (CDC)-type light traps placed at a height of 1.5m above ground.The entomological study was carried out on new moon days during the period March 2013 to February 2015 with collections performed from 1800 to 0600 hours. Phlebotomines were captured in the vicinity of 20 houses selected at random in the urban neighborhood of Salobro . All phlebotomines were identified according to the Young and Duncan  classifi5'-GTGGCCGAACATAATGTTAG-3' and 5'-CCACGAACAAGTTCAACATC-3' to amplify a 200 base pair (bp) fragment from the cacophony gene IVS6 region in New World sandflies [Lu. longipalpis, and a negative control comprising a reaction mixture without DNA, were used in the amplifications.DNA was extracted using the Gentra Puregene Blood Kits , used according to the manufacturer's instructions, and extracts were stored at -20°C until required for analysis by polymerase chain reaction (PCR). In order to serve as a control for the extraction procedure, and to exclude the possibility of PCR inhibition, DNA samples were submitted to PCR analysis using the primers andflies . A positLeishmania DNA by amplification of a 300–350 bp fragment of the internal transcribed spacer 1 (ITS1) using the primers 5’CTGGATCATTTTCCGATG3’ (LITSR) and 5’TGATACCACTTATCGCACTT’3 (L5.8S) [HaeIII digestion) and the band patterns were compared with those obtained from the DNA of the World Health Organization Leishmania strains reference L. amazonensis (IFLA/BR/1967/PH8), L. guyanensis (MHOM/BR/1975/M1176), L. braziliensis (MHOM/BR/1975/M2903), L. lainsoni (MHOM/BR/81/M6426), L. naiffi (MDAS/BR/1979/M5533), L. shawi (MCEB/BR/1984/M84408), L. mexicana (HMOM/BZ/1982/BEL21) and L. infantum (MHOM/1973/BH46), which had been characterized in the Laboratory of Leishmania Biology, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais.DNA samples was screened for the presence of  (L5.8S) ,23. PCR  (L5.8S) , and the (L5.8S) . The ITS™ PCR beads  and primers described previously [cytB) gene. The cytB-PCR amplicons were submitted to RFLP analysis (HaeIII and MboI digestion). Amplification and restriction products were subjected to electrophoresis on 5% polyacrylamide gel [Didelphis albiventris) , pig (Sus domesticus), cattle (Bos taurus), human (Homo sapiens), rodent (Mus musculus), dog (Canis familiaris), chicken  and horse  .In order to test for the presence of DNA from the blood of vertebrate hosts, samples of sandfly DNA were submitted to PCR analysis using PuRe Taq Ready-To-Goeviously  for the mide gel , and themide gel . The banwww.labimage.net, was employed. This software is able to determine the size of each DNA fragment (band) selected in the gel constitutive of each DNA band in comparison with the fragments of the 25 bp and 50 bp DNA ladder .To assure reproducible results, all reactions were performed at least in three independent experiments. The gels were carefully analyzed to determine the PCR-RFLP restriction patterns. To perform data analysis the LabImage-1D Gel Analysis software, Version 2.7.2 , available at the website The 200 bp-fragment from the cacophony IVS6 region of phlebotomines could be amplified in all of the sandfly DNA samples studied, thus confirming the high quality of the extracted DNA and the absence of PCR inhibitors.Lu. longipalpis (n = 763), Lu. evandroi (n = 3), Lu. sordellii (n = 1), Lu. lenti (n = 1) and Lu. whitmani (n = 1), while 213 were collected in the rural area and comprised the species Lu. longipalpis (n = 175), Lu. whitmani (n = 23), Lu. trinidadensis (n = 6), Lu. termitophila (n = 5), Lu. lenti (n = 3) and Lu. sordellii (n = 1).Of the 982 female phlebotomines dissected, 769 were collected in urban settings and comprised the species Leishmania sp. presenting an overall infection rate of 5.2% (Leishmania identified by ITS1-PCR-RFLP were L. infantum (n = 28), mixed L. infantum/L. braziliensis (n = 8), L. braziliensis (n = 3), L. shawi (n = 3), Leishmania sp. (n = 3), L. mexicana (n = 2), L. guyanensis (n = 2), L. amazonensis (n = 1) and L. lainsoni or L. naiffi (n = 1) , but rates varied substantially among species of sandfly with Lu. termitophila presenting the highest frequency of infection (40.0%), followed by Lu. trinidadensis (33.0%). Lu. whitmani (17.0%) and Lu. longipalpis (11.0%). Infection by L. infantum and mixed infection by L. infantum/L. braziliensis were the most frequent.According to ITS1-PCR analysis, 51 of the 982 female sandflies were positive for  of 5.2% , Fig 2A. (n = 1) . In the cytB-PCR-RFLP band patterns following MboI and HaeIII digestion (Sources of blood feed could be identified in 73.7% (573/778) of engorged female sandflies, most of which were collected in the urban area . The Considering sandflies that had simple feeding , food soWith regard to sandflies that had fed on two hosts (mixed feeding) , prefereLu. longipalpis fed on the blood of all of the vertebrate hosts studied but showed a single source preference for, in decreasing order, chicken, dog, rodent and human. Furthermore, this phlebotomine fed on eight host combinations in the urban area and five host combinations in the rural area, with the predominance of dog/chicken followed by dog/rodent. In the rural area, Lu. whitmani exhibited a single source preference for human, dog, chicken, and rodent blood, in decreasing order, and for dog/rodent and dog/chicken combinations. The only example of an engorged Lu. whitmani female captured in the urban area has fed on a dog/rodent combination. The single source preferences of Lu. evandroi and Lu. lenti in the urban area were dog and rodent, respectively, while dog was the preferential host for Lu. termitophila, Lu. trinidadensis and Lu. sordellii in the rural area. A dog/rodent combination was established as the blood source for Lu. trinidadensis, Lu. termitophila and Lu. lenti that had fed on two hosts in the rural area. As shown in Leishmania-infected blood in engorged sandflies were dog, rodent and human, independent of the number of vertebrate hosts or Leishmania species.Females of Leishmania infection in the canine reservoirs coupled with the L. infantum-infection rates in Lu. longipalpis (AVL vector) of 2.0 and 7.0% in urban and rural areas, respectively, and the L. braziliensis-infection rate in Lu. whitmani (ATL vector) of 4.0% [Between March 2013 and February 2015, 107 human and 2,380 canine cases of AVL were recorded in the municipality of Caxias, together with 62 human cases of ATL [ of 4.0% ,27,28.L. infantum infection was detected in the AVL vector Lu. longipalpis and in non-vectorial species such as Lu. whitmani and Lu. termitophila. The natural infection of Lu. longipalpis by L. infantum has been described in various AVL-endemic regions of Brazil [Lu. longipalpis and of infected canine reservoirs, which are widely distributed in the State [In the area encompassed by the present study, f Brazil –33, inclf Brazil , and thef Brazil . Indeed,he State .Lu. longipalpis by L. infantum is low, even in areas with high AVL endemicity. However, based on the high degree of anthropophilia, vectorial capacity, simultaneous presence of vector and disease, and the abundance and spatial distribution of the vector observed in the present study, it is possible to state with certainty that Lu. longipalpis plays a key role in the AVL transmission cycle in the municipality of Caxias where prevalence rates of human and canine cases are elevated.In nature, the infection rate of Lu. whitmani and Lu. termitophila by L. infantum was detected for the first time in the rural area of Caxias during the present study. This finding is important since Lu. whitmani was the second most abundant species of sandfly in the study area, particularly in the rural setting. On the other hand, the high rate of infection (20%) of Lu. termitophila by L. infantum must be examined carefully since only one specimen was infected. Natural infection of Lu. whitmani and Lu. termitophila by L. infantum was first recorded by Saraiva et al. [L. infantum naturally infecting the phlebotomines Lu. migonei and Lu. cortelezzii, both species of which are vectors of ATL in Brazil while the latter is a proven vector of ATL in Argentina [Infection of a et al.  in the srgentina –38.L. infantum in sandflies that are not considered to be vectors of AVL demonstrates the ability of a range of phlebotomine species to acquire L. infantum infection. The importance of these parasite-phlebotomine associations, even if they occur only occasionally, requires further study in order fully to understand their implications regarding the ecoepidemiology of leishmaniasis. However, the conclusion that a species of sandfly is a vector in the leishmaniasis cycle cannot be based exclusively on the occurrence of natural infection by the causative agent. Together with the natural infection factor, a vector of Leishmania sp. must fit the criteria described by Killick-Kendrick [The finding of infection by Kendrick .Lu longipalpis naturally infected by L. braziliensis, L. amazonensis, L. mexicana, L. shawi, L. guyanensis and L. lainsoni or L. naiffi in Maranhão, in an area of cerrado that is not within the Amazon region. However, it is important to emphasize that Lu longipalpis was the most abundant and widespread sandfly species in the study area and appeared to be perfectly adapted to domestic and anthropic settings.We report here for the first time the occurrence of Lu longipalpis naturally infected by L. braziliensis has been described in other regions of Brazil [Lu. longipalpis naturally infected by L. amazonensis has been established in the state of Mato Grosso do Sul [L. amazonensis is the causative agent of ATL, including the anergic or diffuse cutaneous forms of the disease, and its vectors are normally Lu. flaviscutellata and Lu. olmeca in the Brazilian Amazon region [L. amazonensis have been reported in all geographical and ecological regions of Maranhão [The presence of f Brazil ,40, whilo do Sul ,42. L. an region . These tMaranhão ,45.Lu. longipalpis infected by L. mexicana has not been previously reported in Brazil, L. mexicana-infected humans have occasionally been recorded [Lu. longipalpis naturally infected by L. shawi, neither in Maranhão nor in other Brazilian regions, suggesting that this phlebotomine does not form part of the L. shawi transmission cycle outside the Amazon region, where the normal vector is Lu. whitmani. Although we found Lu. longipalpis infected by L. guyanensis in the rural area of Caxias, this species of Leishmania is usually restricted to recently colonized areas of the Amazon region, where Lu. umbratilis and Lu. anduzei are the main vectors [L. guyanensis has also been detected in Minas Gerais in a pool of sandflies that are not considered to be vectors of ATL [While the occurrence of recorded . There a vectors  and eden vectors ,48. The s of ATL .L. lainsoni, or possibly L. naiffi, is reported here for the first time outside the Amazon region of Brazil. The main vector of L. lainsoni in the Amazon is Lu. ubiquitalis, although there are reports of the occurrence of L. lainsoni-infected Lu. davisi in the state of Amazonas [L. lainsoni-infected Lu. nuneztovari anglesi in Bolivia [L. lainsoni in South America, which has now been shown to extend to Caxias in the state of Maranhão. The main vectors of L. naiffi, a Leishmania generally found in the Amazon region, are Lu. squamiventris, Lu. paraensis and Lu. ayrozai [Lu. davisi and Lu. hirsuta hirsuta, none of these sandflies have been identified in the Caxias area [The presence of Amazonas –52 and o Bolivia . Moreove Bolivia . These f ayrozai . Althougias area ,57.L. braziliensis-infected Lu. whitmani was confirmed at the site of the present study. This phlebotomine is a natural vector of L. braziliensis in various Brazilian regions [L. braziliensis is distributed throughout the ATL-endemic zone of Brazil, from north to south of the country and in both new and well-established settlements, and is commonly associated with the presence of a range of domestic, synanthropic and wild animals.The presence of  regions , especia regions ,28, and  regions , generalLu. longipalpis, Lu. whitmani and Lu. trinidadensis co-infected by L. infantum/L. braziliensis, examples of which have not been recorded recently, is reported herein for the first time in Caxias. There is, however, no evidence that Lu. whitmani and Lu. trinidadensis are involved in the transmission of L. infantum or that Lu. trinidadensis plays a role in the transmission of L. braziliensis to humans. Although reports of mixed infections are available, including that of Lu. flaviscutellata by L. braziliensis and L. amazonensis [Leishmania are more common than the literature would suggest, particularly in foci where the two species overlap.The presence of zonensis , we beliLeishmania sp., which can be attributed to the presence of unidentified species of Leishmania or other trypanosomatids. These problems could not be resolved using the PCR-RFLP technique owing to the small amount of DNA available or to the presence of degraded DNAIn the present study, three sandfly females were classified as infected by Leishmania transmission, interactions between Leishmania vectors and reservoirs and the anthropophilic attraction of vectors [L. donovani transmission areas has been associated with increased risk to humans in some studies and with decreased risk in others, showing that the relationship between mammals and phlebotomines is complex and can affect the abundance, aggregation, feeding behavior and infection rates of the vector [The blood food sources of female sandflies were identified in the present study since this information can improve our knowledge regarding  vectors ,61. Furt vectors . For exae vector .Lu. longipalpis was the most opportunistic species, since it fed on the blood of all vertebrate hosts tested, but exhibited a preference for chickens, dogs rodents and humans, and, to a lesser extent, pigs and horses. Chickens were also the preferential source of blood for female sandflies when all identified species were considered together. Chickens are very common in rural and urban settings, and chicken coops represent attractive areas for phlebotomines to rest and reproduce [Leishmania, they may be important in the maintenance and domiciliation of vectors by attracting mammalian reservoirs to the vicinity [Leishmania has already been detected in Maranhão [In the present study, eproduce –66. Althvicinity . It is pvicinity . Galati vicinity  called aMaranhão .Although horses were not high on the preference list of phlebotomine food sources, the intense movement of these animals between rural villages, and even in the urban periphery, could be a risk factor for leishmaniasis. Indeed, clinical cases of naturally acquired leishmaniasis have occasionally been found in horses in transmission areas in Brazil, specifically São Paulo  and MinaLeishmania vectors [Considering mixed blood feeding, the dog/rodent combination was the most frequent blood source for female sandflies. This type of feeding behavior reflects the adaptability of sandflies and indicates that these insects can adjust their feeding patterns according to availability, accessibility, abundance, size and biomass of vertebrate hosts . Under c vectors .Lu. longipalpis and Lu. whitmani were found to be infected by L. infantum and L. braziliensis, and Lu longipalpis and Lu. trinidadensis were co-infected by L. infantum/L. braziliensis. We also observed that, occasionally, Lu. evandroi and Lu. sordellii performed single feeding on dogs and Lu. lenti fed on rodents, either alone or in combination with dogs, but without infection by Leishmania. From an epidemiological viewpoint, it would appear that these species of phlebotomine are not involved in the Leishmania transmission cycle in Caxias.Preferences for dogs, rodents and humans, along with the observed mixed feeding behavior, constitute an important epidemiological factor in Caxias since, in our study, L. infantum. Since infection rates are commonly high among canine populations in endemic areas, and a large proportion of affected animals are asymptomatic [L. braziliensis in Brazil [L. braziliensis in dogs has been amply described [L. amazonensis [L. infantum/L. braziliensis [Leishmania that cause ATL are always lower in dogs than in humans. It may be that dogs, similar to humans, are victims of the disease and not just carriers of parasites in nature.Among domestic animals, dogs are the species most commonly affected by leishmaniasis and have long been recognized as the main domestic reservoirs of ptomatic , dogs reptomatic . Furthern Brazil –78, evenescribed –79, and zonensis  or co-iniliensis  have beeL. braziliensis in nature [Leishmania sp. since they represent an important link between domestic and wild environments [Results of a study conducted in northeastern Brazil strongly indicated that some small rodents are involved in the maintenance of n nature , reinforronments .Lu longipalpis fed on the blood of opossum in the peridomicile, even at low frequency, is most interesting because it establishes a connection between the domestic and wild cycles of AVL and, according to Cabrera et al. [Our observation that a et al. , may givLu. longipalpis and Lu. whitmani feed on human blood as well as on the blood of one or more hosts  that inhabit the environs of rural and urban dwellings. Such findings provide strong evidence that these phlebotomines exhibit anthropophilic behavior and support our original hypothesis that, in the municipality of Caxias, transmission of Leishmania occurs in close proximity to humans (i.e. inside or near domiciles). However, the observations reported herein of natural infection of non-vectorial sandflies by Leishmania sp. do not prove the participation of such species in the transmission of AVL and ATL in Caxias.In conclusion, the present study has demonstrated that Leishmania, with the potential to cause different clinical manifestations, which can be transmitted by different vectors and maintained by several mammal reservoirs. In spite of the high number of human cases and considering that current control and prevention methods have not been shown to be effective, new strategies for appropriate intervention and control should be developed to avoid propagating even to non-endemic areas. From this perspective, has become increasingly important to study essential issues involved in the transmission, such as the natural sand fly infection rates of the vectors by Leishamania sp., the geographical distribution of these parasites in potential vectors and the study of their blood meal sources, which are important tools for eco-epidemiological studies of leishmaniasis.In Brazil, the transmission cycles of leishmaniasis have been found to have a focal distribution in specific geographic areas. Leishmaniasis is characterized by complex and specific epidemiological profiles for each local of transmission, involving several species of Our findings have important implications relating to the epidemiology of leishmaniasis especially regarding vector competence, knowledge of which is vital for successful vector surveillance and control.S1 Table(XLSX)Click here for additional data file.


Despite the significant progress made in prevention, diagnosis, and treatment, there is still a high rate of vaginal dysbiosis in Kazakh women. The use of antibiotics in the treatment of vaginal dysbiosis contributes to the elimination of pathogens as well as microflora, which can lead to a decrease in local immunity and more favorable conditions for infection spread. The most physiologically safe and promising method for the restoration of vaginal biocenosis is the use of probiotics administered by a vaginal route.We have allocated 64 of cultures of Lactobacillus from the vaginal epithelium of healthy women of reproductive age and women with diagnosed bacterial vaginosis (BV). Identification of cultures was performed by PCR analysis of 16S ribosomal RNA. Evaluation of biological significance was determined by the following criteria: high antagonistic activity against Candida albicans, Escherichia coli, Serratia marcescens, Proteus mirabilis, Klebsiella ozaenae, and Staphylococcus aureus; and production of hydrogen peroxide, resistance to antibiotics, adhesive activity. We studied the symbiotic relationship of selected biologically active of cultures to each other and received options for consortiums with properties of probiotics through co-cultivation.Results of genotyping showed that the isolated lactobacilli belong to the seven species: L. fermentum, L. salivarius, L. gasseri, L. crispatus, L. jensenii, L. plantarum, and L. delbrueskii. L. fermentum, L. salivarius, L. gasseri, and L. jensenii occur in women with suspected BV. The highest percentage of occurrence in the vagina of healthy women was L. fermentum (28%). Most strains of lactobacilli possess high inhibitory activity for all test-strains, except Candida albicans (37.5%). 56% of studied cultures revealed high adhesion to human erythrocytes. All lactobacillus strains were resistant to metronidazole, 80% to kanamycin, 57% to vancomycin, and sensitivity to roxithromycin, amoxiclav, ampicillin was diagnosed in all strains. 50% of cultures showed a moderate sensitivity to gentamicin and cefazolin. In a study of peroxide-producing activity, 80% of the cultures exhibited peroxide-producing activity. As a result of screening, the 7 most active strains of lactobacilli were selected for development of 10 variants of probiotic consortia. Also, there was increase of adhesive activity in the consortia compared to other components. These consortia can be used for the treatment of BV in addition to metronidazole.The probiotic consortia identified in this study had high antagonistic, adhesive properties, and resistance to metronidazole. These probiotics can potentially be used for the development of biological products for the treatment and prevention of bacterial vaginosis.


Mosquito nets containing synergists designed to overcome metabolic resistance mechanisms in vectors have been developed. These may enhance excitability in the mosquitoes and affect how they respond to CDC light-traps. Investigating the behaviour of vectors of disease in relation to novel mosquito nets is, therefore, essential for the design of sampling and surveillance systems.® long lasting insecticidal net (LLIN) and Olyset Plus® LLIN containing piperonyl butoxide (PBO) synergist. Nets were rotated daily between the nine rooms over nine nights. A further series of experiments using the nets on alternate nights in a single room was undertaken during the short rains. Anopheles gambiae s.l. were collected in CDC light-traps, a window-trap and Furvela tent-trap. Anopheles gambiae s.l. were identified to species by polymerase chain reaction (PCR).In an initial experiment in Muleba, Tanzania, nine bedrooms from three housing clusters were sampled. CDC light-traps were operated indoors next to occupied untreated nets (UTN), OlysetAn. gambiae s.l. were An. gambiae s.s., the remainder being Anopheles arabiensis. The number of mosquitoes collected from 81 light-trap collections was greater in the presence of an Olyset  relative to an UTN. In a second experiment, in the wet season 84% of the 180 An. gambiae s.l. identified were An. arabiensis. The number of An. gambiae s.l. collected from a light-trap compared to a tent-trap was significantly higher when an Olyset Plus net was used compared to an UTN. Survival of the mosquitoes in the window trap was not reduced by the use of an Olyset Plus net in the bedroom relative to an Olyset net.In the initial experiment 97.7% of the 310 An. gambiae or An. arabiensis collected in light-traps when a treated net is used requires further experimentation and may be because of a heightened escape reaction on the part of the mosquito.Mosquitoes entering bedrooms, even those susceptible to pyrethroids, were not killed by contact with an Olyset Plus LLIN. The enhanced numbers of  Anopheles arabiensis collected in households with light-traps in the presence of deltamethrin-treated or UTNs, however, reported no significant difference in numbers caught . There was no significant difference (p > 0.05) between the numbers of C. fuscopennata .Three-hundred-and-three (97.7%) of the 310  gambiae . In the An. arabiensis was relatively stable. The proportion of engorged and part-fed female mosquitoes was significantly different when sampling was performed with an UTN relative to an Olyset or Olyset Plus . Despite the provision of sugar solution and usage of a conical collection bag, only 10% of the mosquitoes were alive in the morning. The proportion of An. arabiensis collected alive or dead was not affected by the type of net in use . The total nightly indoor collection (light-trap and window-trap), which was always small compared to the light-trap, was also independent of the net in use.During the sampling conducted during the short rains, the population of An. arabiensis were collected in the light-trap compared to the tent-trap  whereas the numbers collected when an Olyset LLIN or UTN was present were not significantly different  and , respectively  An. gambiae in the dry season and susceptible An. arabiensis in the wet season, were caught when an UTN was used compared to the standard Olyset or Olyset Plus. If, as seems reasonable based on previous hut trials in Benin [An. gambiae s.l. entering houses, deterrent effect [The results obtained from both sets of experiments indicate that the type of mosquito net in use affected the numbers of mosquitoes caught by light-traps. Surprisingly, significantly fewer mosquitoes, both resistant in Benin , the nett effect , and theAn. arabiensis population was susceptible to pyrethroids [An. gambiae in Muleba is highly resistant to this group of insecticides [The results of the bioassay conducted with the standard Olyset in the wet season indicated that the ethroids . On the cticides , 14, 27.In many areas people may possess LLINs but may not use them. Non-LLIN-using households may be at a greater risk when PBO-containing nets are used by their neighbours, as they may be when topical repellents are used . Distrib


AbstractPtychadena of the herpetological collection of the Museo Nacional de Ciencias Naturales de Madrid reveals the first record of Ptychadenaschillukorum from Mauritania, extending the known distribution range of the species in West Africa more than 450 km northwards. A key is provided for the four Mauritanian species of Ptychadena to solve problems in identification encountered in previous studies.The study of specimens of the genus  Ptychadena Boulenger, 1917 is a genus of medium-sized Afrotropical frogs that comprises 50 known living species , P.bibroni , P.tellini  ), P.longirostris , P.oxyrhynchus , P.trinodis , P.mascareniensis , P.tournieri , P.stenocephala , P.submascareniensis , P.retropunctata , and P.schillukorum  ) ) . At the bibroni . PtychadPtychadenatrinodis differs from the other two species of Ptychadena recorded in Mauritania by the presence of a tarsal tubercle and by very large inner and outer metatarsal tubercles; in Mauritania it has been reported in two localities: Guelta Oumm Lebare and Guelta Metraucha. Ptychadenacf.mascareniensis is cited only at one locality in Mauritania: Zoueina; it differs from P.bibroni by a very subtle outer metatarsal tubercle, lack of supernumerary tubercles on toe IV, dorsal median ridges that reach the posterior edge of the eyes, different webbing formula (22/3–3 in P.cf.mascareniensis and 2–21/3 in P.bibroni), and by a continuous lateral ridge which is broken in P.bibroni. Ptychadenabibroni is known by two records in Mauritania: Kaedi and Mahmoûdé Lake PtychadenPtychadena). Only Hoplobatrachusoccipitalis, Sclerophrysxeros, and Tomopternacryptotis, are present across the north in the Saharan realm  and on field data obtained by RM.In this note, a new species record of Ptychadena held at the National Museum of Natural Sciences of Madrid (MNCN-CSIC) were examined. Thirty-one specimens were collected in Mauritania: one specimen of Ptychadenatrinodis  were collected in October 1998 by RM, a few days after a (rare) event of rainfall in Southern Mauritania in the proximities of Aleg . Additionally, 10 individuals (MNCN 43846–43855) were collected in October 2006 by J. M. Padial and I. De la Riva in Aleg surroundings  . In this work the criteria of Padial and De la Riva considering the presence of P.cf.mascareniensis in Mauritania are followed.The poor conditions in which we found the specimen of  la Riva  makes it la Riva . Unfortu la Riva  of the sPtychadenaschillukorum from Mauritania fitted with the description of the species  event of rainfall that caused the formation of shallow pools and the growth of herbaceous vegetation on the normally barren sand Fig. . Other atis Fig. .Ptychadena  while in P.trinodis a fine white-yellow vertebral line and light colored lateral ridge is generally present. Ptychadenaschillukorum differs from P.bibroni by its more compact body, shorter legs, lack of supernumerary tubercles on toe IV, always interrupted dorsal ridges, by its inconspicuous outer metatarsal tubercle and by thigh pattern, that is speckled in P.schillukorum and usually with yellow longitudinal and irregular line in P.bibroni. Finally, P.schillukorum is distinctive from P.mascareniensis by its compact body shape, dorsal and lateral ridges, always interrupted, and shorter legs.The four Mauritanian species of a Figure  are easiPtychadena species that occur in Mauritania is presented.Based on Ptychadenaschillukorum may be discovered in the future. Molecular studies of P.schillukorum should be carried out to know how complex is the species across their distribution (expected to be high or a group of species as suggested by Studies concerning amphibian fauna in Mauritania are scarce, suggesting that other populations of PageBreak


Papuanella  was established by Papuanella is currently recognized as a valid genus within the subfamily Papuininae of the family Camaenidae . Species included in the genus as defined by O.ogeramuensis , comb. n., and O.finisterrensis , comb. n.nclature , the namOgeramua is named for Ogeramua, Papua New Guinea, the type locality of its type species, and is to be treated as feminine in gender.The epithet


Dr. Navnendra Mathur (28.09.1951 - 03.07.2009)What does one say about a dedicated academician, an astute clinician; a versatile professional well versed in Orthopedics, Physical Medicine and Rehabilitation and a great endearing colleague and friend!Dr. Navnendra Mathur did his MS Ortho from SMS Medical College, Jaipur, in 1980. He then did his DNB in Physical Medicine and Rehabilitation in 1986. The following year he was selected for the prestigious IOA Johnson and Johnson Fellowship. After a stint as orthopedic specialist at King Fahd Hospital, Jeddah, Saudi Arabia he returned to join as Assistant Professor in Physical Medicine and Rehabilitation. He rose to the post of Professor and Head of Department, Physical Medicine; Director of the Rehabilitation Research Centre set up by his mentor Dr PK Sethi.Dr. Navnendra was virtually obsessed with the study of spinal injuries and presented many outstanding papers on the subject at national and international conferences including the International Spine and Spinal injuries Conference at Indian Spinal Injuries Centre, New Delhi; International Society of Physical Medicine and Rehabilitation Medicine, Seoul, Asian Oceana Conference of Physical Medicine & Rehabilitation, Nanking, China and ASCON at Vietnam last year. He was also slated to attend the International Spine meet at Hong Kong this month. A life member of IOA, Dr Mathur has to his credit more than 25 publications in international and national Orthopedic and Physical medicine journals.Born in November 1951, Dr. Navnendra had yet to complete 58 years. He passed away on July 3, 2009. He is survived by his wife Dr. Sushma, a CGHS GDMO, and two daughters.


Anaplasma phagocytophilum in Europe is transmitted by the ixodid tick vector Ixodes ricinus. A. phagocytophilum causes a disease with diverse clinical signs in various hosts. A great genetic diversity of the groESL operon of A. phagocytophilum has been found in ticks elsewhere. In Slovenia, the variety of the groESL operon was conducted only on deer samples. In this study, the prevalence of infected ticks was estimated and the diversity of A. phagocytophilum was evaluated. On 8 locations in Slovenia, 1924 and 5049 (6973) I. ricinus ticks were collected from vegetation in the years 2005 and 2006, respectively. All three feeding stages of the tick's life cycle were examined. The prevalence of ticks infected with A. phagocytophilum in the year 2005 and in the year 2006 was 0.31% and 0.63%, respectively, and it did not differ considerably between locations. The similarity among the sequences of groESL ranged from 95.6% to 99.8%. They clustered in two genetic lineages along with A. phagocytophilum from Slovenian deer. One sequence formed a separate cluster. According to our study, the prevalence of A. phagocytophilum in ticks is comparable to the findings in other studies in Europe, and it does not vary considerably between locations and tick stages. According to groESL operon analysis, two genetic lineages have been confirmed and one proposed. Further studies on other genes would be useful to obtain more information on genetic diversity of A. phagocytophilum in ticks in Slovenia.Ticks act as vectors of many pathogens of domestic animals and humans.  Ixodes ricinus [I. ricinus feeds on livestock, deer, dogs and a wide variety of other species, including humans [I. ricinus is a confirmed vector of the bacterium Anaplasma phagocytophilum [A. phagocytophilum, the agent of granulocytic anaplasmosis, was formerly known as human granulocytic ehrlichiosis agent (HGE agent), Ehrlichia phagocytophila and E. equi [A. phagocytophilum causes a disease with diverse clinical signs in various hosts from asymptomatic to life-threatening [A. phagocytophilum, the groESL operon is widely used as the 16 S rRNA gene is too conservative [groESL sequences of A. phagocytophilum in ticks has also been described in Germany [A. phagocytophilum was estimated and the diversity of the groESL operon of detected DNA of A. phagocytophilum was evaluated.Ticks and tick-borne diseases affect animal and human health worldwide. A vector of many diseases in Europe and Slovenia is  ricinus . It can  ricinus . Ticks' g humans . I. ricitophilum . The tictophilum . A. phag E. equi . A. phageatening . No fataeatening . Importaeatening . The wileatening . The preeatening . To descervative . It has ervative . An imme Germany . The var Germany . In a pr Germany . The tic2 over 100 m of vegetation [16 S rRNA was examined [16 S rRNA of genus Anaplasma sp. and Ehrlichia sp., was used [groESL operon. A nested PCR would amplify a 1296-bp fragment of groESL operon of A. phagocytophilum variants [A. phagocytophilum, grown in a HL-60 cell culture, was used as a PCR positive control. If a pool of adult ticks was positive, the stored half of each dissected tick from a pool was used for DNA extraction and further amplifications. All amplicons of groESL operon were further analyzed by sequencing on both strands with the BigDye Terminator Cycle Sequencing Ready Reaction Kit . The sequences were analyzed with computer programs of the Lasergene 1999 software package  based on Clustal W algorithm [® Excel Add-In, developed by Brad Biggerstaff, CDC, Fort Collins, CO). Statistical analyses were performed using SPSS version 17.0 . P values of 0.05 or less were considered statistically significant.The study was performed in the years 2005 and 2006 at 8 locations in Slovenia. The criteria for selecting the locations were the tick-borne pathogens' presence in human patients or a higher altitude of the location compared to others. Ticks were collected at forest edges by dragging a flag with a surface of 1 mexamined . For thewas used . All poslgorithm . The dislgorithm  and the lgorithm  was usedI. ricinus ticks were collected by flagging vegetation in the years 2005 and 2006, respectively. Ticks were separated into pools: 252 pools in 2005 and 442 pools in 2006  p. ticks , only I.16 S rRNA of Anaplasma sp. was detected in 26 pools of adult and nymphal stages of ticks . Probably, a novel genetic lineage of the groESL operon of A. phagocytophilum was found. To obtain more information about genetic diversity of A. phagocytophilum in I. ricinus in Slovenia, additional genetic markers, such as ankA and msp4, should be analyzed.In one pool of nymphs from Mozirje, a I. ricinus nymphal and adult stages are responsible for the transmission of the pathogen as A. phagocytophilum was present in both stages. There was no significant difference in the prevalences of A. phagocytophilum at different locations and in both years. The prevalence of infection in ticks did not differ considerably from the reports from elsewhere in Europe. With this study, we have confirmed that I. ricinus is a vector of a variant of A. phagocytophilum that causes the disease in humans and dogs. The sequencing of the groESL operon has demonstrated a great diversity of A. phagocytophilum in Slovenia. With phylogenetic analysis, two genetic lineages have been confirmed and another has been proposed. Further phylogenetic studies of several other genes, such as ankA and msp4, might be useful to obtain more information about genetic diversity.The authors declare that they have no competing interests.KSS conducted the laboratory study and drafted the manuscript. MS was involved in laboratory study. DD facilitated the molecular laboratory study. NK conducted the field study and did the statistical analysis. TAZ was involved in the project design and participated in drafting the manuscript. All co-authors have read the manuscript.


He studied at Carmichael Medical College, Kolkata, and completed his M.B.B.S. in 1952. He joined Patna University in 1954 for a postgraduate degree in General Surgery and graduated with an M.S. (Ortho) degree from Patna in 1957.Dr. Mahendra T. Mehta was the first Orthopedic Surgeon of Gujarat and the President of the Indian Orthopedic Association from 1988 to 1989. He was born in Zariya, Bihar on February 5His first research project, 'Diaphysecomy and Bone Regeneration' initiated in Patna in 1955, was supported by a scholarship from Lady Tata Memorial Trust. He joined Sheth Vadilal Sarabhai Hospital in 1957 and later in 1961, joined Civil Hospital, Ahmedabad as an Honorary Orthopedic Surgeon. He continued his research here with a research grant from Gujarat University, and finally presented his work at the SICOT meeting in Copenhagen in 1975.He had a great interest in C.T.E.V and considered manual correction and maintenance with a plaster cast to be a mainstay in its management. Armed with the training imparted by his mentor, Dr. B. Mukhopadyay, he started anterolateral decompression for Pott's Paraplegia in 1957 in Ahmedabad. In the late 1950s, he gave up immobilization and accepted active mobilization for tuberculosis of joints. He was awarded the B. N. Mehta Oration Award in 1974 after his presentation of his work on bone and joint tuberculosis.He was a teacher to hundreds of students, yet his humble words to his family in the last week of his life were, 'I am a student and I have yet to learn a lot.' Not only was he a host for many medical students, doctors, and visiting orthopedic surgeon friends, but he also looked after all Johnson and Johnson fellows during their visits to Ahmedabad. Family members concede: 'He taught us to transform relations to dear ones and dear ones to great relationships'. His family has lost a most dear one, but feels his presence all the time, even after his death.He offered deep insights into orthopedics as a member of the Advisory committee appointed by the IOA in 1980 for the standardization of a postgraduate (PG) degree in orthopedics in India. The committee brought in the idea of a PG work book, a precursor of the log book. He served B. J. Medical College and Civil Hospital as an Honorary Professor of Orthopedics from 1961 to 1981, and served various hospitals in Ahmedabad, including the Cancer Hospital in Ahmedabad.He was a recipient of P. R. Trivedi (1980) and Dr. Katrak (1982) Oration Awards. He was the founder President of the Gujarat Orthopedic Association. He published, 'Forty years of Orthopedics—As I Saw It' in February 1998. It's a small book, but a treasure of precious thoughts.He has made a great difference for many lives, not only as an orthopedic surgeon, but also as a great human being. In his death on August 14, 2008, Yasubhabhi has lost a partner who greatly loved her and always took her with him on all his travels. In his death, we orthopedic surgeons in Gujarat have lost a father, who kept all of us together in our nest.May God Almighty rest his soul in Eternal Peace.


Amaranthus cruentus L.  and nightshade, Solanum macrocarpon L.  were examined on the development and demographic parameters of the spider mite, Tetranychus ludeni Zacher (Acari: Tetranychidae). This mite was recently identified as a pest of the two leafy vegetables which are widely used in West Africa. The experiments were conducted at the International Institute of Tropical Agriculture, Benin, West Africa, in a growth chamber at 27°C, 70% ±10% RH and 12:12 (L:D). Immature development of T. ludeni was shorter on A. cruentus than on S. macrocarpon, whereas female longevity was the same on the two vegetable crops. Total fecundity per female was higher on A. cruentus than on S. macrocarpon, largely due to longer survival of adult female T. ludeni on the former; however, no differences were observed in the daily fecundity of T. ludeni on the two plant species. The comparison of intrinsic rates of natural increase (mr), the net reproductive rates (oR) and the survival rates of adult stage of T. ludeni on the two vegetable crops suggests that T. ludeni performs better on S. macrocarpon than on A. cruentus. Reasons for the lower rate of population growth observed on amaranth should be studied in more details as this could be used in IPM strategies such as intercropping to reduce pest density and in developing biopesticides for use against T. ludeni in vegetable farms in Africa.It is well recognized that the quality of host plants affects the development and survival of plant-feeding arthropods. The effects of two leafy vegetable crops, amaranth,  It is now well recognized that host plant quality can affect several life-history characteristics of their herbivores, by impairing growth, lowering resistance to disease and reducing fecundity . AlteratIn the prevailing vegetable cropping systems in sub-Saharan Africa, several vegetable crop species are grown in close proximity in small lots. The crops are attacked by several pest species , and theTetranychus urticae Koch and T. ludeni Zacher are the most important on many vegetable crops  in the family Tetranychidae are among the most important crop pests worldwide. The subfamily Tetranychinae includes a number of economically significant species of which le crops . These tle crops . Tetranyorldwide . In Indiand okra . Controland okra , which hand okra , observele farms  with theTo overcome the problems of resistance development in spider mites, the search for more durable crop protection solutions based on IPM systems in vegetable farms is necessary . HoweverReproduction in spider mites is very sensitive to a wide variety of intrinsic and extrinsic conditions. Intrinsic factors that influence life table parameters of tetranychid mites include mite strain and level of inbreeding, colony density, age of females and of the population, fertility status of the females and various behavioural parameters . ExtrinsAmaranthus emeritus L.  and nightshade, Solanum macrocarpon L. , locally known as ‘Gboma’, are respectively the first and second most commonly grown and consumed leafy vegetable crops species in southern Benin , in a field at the International Institute of Tropical Agriculture (IITA), Cotonou Station in Benin. These mite colonies were maintained indoors on potted plants of S. aethiopicum at 26 ± 1°C and 65–85% RH for 2 weeks  cut from S. aethiopicum were transferred singly using a camel hair brush onto fresh leaf discs (22 mm in diameter), cut from each of A. cruentus and S. macrocarpon, respectively. These leaf discs were kept on water saturated cotton wool in Petri dishes (15 cm in diameter). Water was added each day as needed to ensure adequate moisture for the leaf discs and to maintain a water barrier to restrict mite movement. These rearing units were kept in a growth chamber at 27°C, 70% ± 10% RH and 12:12 (L:D). Leaf discs were replaced twice weekly to maintain freshness of leaves, thereby preserving host-plant attributes. Observations were made everyday at 06:00 hours and 18:00 hours and the developmental stage reached by each individual was recorded until all adults emerged. Individuals that escaped from leaf discs were not included in the analysis.To determine the developmental time of immature stages of To determine the life table characteristics of the herbivorous mite on each of the two vegetable crops, 25 deutonymphal  females see  were ranThe Student t-test in SAS  was usedA. cruentus than on S. macrocarpon . Similarly, total duration of development from egg to adult was also shorter on A. cruentus than on S. macrocarpon . On both vegetable crops, the longest stage was the egg that had a developmental time that was generally more than two fold longer than any of the other stages.Except for egg and larval stages that had the same duration on the two vegetable crops, the other life stages were shorter on T. ludeni, and the length of the pre-oviposition and oviposition periods did not differ significantly between the two vegetable crops tested; however, T ludeni had a longer post-oviposition period on A. cruentus than on S. macrocarpon (P < 0.05). The total number of eggs laid per female was lower on S. macrocarpon than on A. cruentus (P < 0.05); however, daily egg production was similar on the two vegetable crops .The longevity of adult female le crops . Egg prole crops . Peak feT. ludeni decreased considerably on A. cruentus after nine days whereas on S. macrocarpon, the decrease began after five days and became more pronounced after 12 days , net reproductive rate (oR), female progeny, and the survival rate at adult stage of T. ludeni differed between the two host plants. Parameters such as mr, oR and female progeny were respectively 1.24, 2.55 and 2.35 fold higher on S. macrocarpon than on A. cruentus. Similarly, the survival rate of immatures to the adult stage was 72% on S. macrocarpon while it was 45% on A. cruentus; and doubling time was shorter on S. macrocarpon than on A. cruentus.Life table parameters are presented in S. macrocarpon, than on A. cruentus. The production of females seems constant on S. macrocarpon with a trough only on day 19, followed immediately by a rapid increase at the end of the experiment. On A. cruentus, the production of females showed an overall decreasing trend with a trough around day 11 after which it reached a peak before decreasing, starting from day 16 to the end of the experiment.The temporal fluctuations of the percentage of female progeny  against T. ludeni feeds, survives and develops on the two vegetables A. cruentus and S. macrocarpon, and that host type can greatly affect its development, fecundity and life-table parameters. Indeed, the results showed that the host plant had substantial effects on the intrinsic rate of natural increase (mr), the net reproductive rate (oR), female progeny, and the survival of the adult stage.Our study shows that Phaseolus vulgaris L., mean duration of developmental stages for female T. ludeni was 9.98 days  ulmi  ulmi . Lewontius) ulmi , and Wreus) ulmi , showed A. cruentus compared with S. macrocarpon, life table parameters such as number of progeny per female , survival of immatures to the adult stage, proportion of females in the progeny and net reproductive rate were higher on S. macrocarpon than on A. cruentus. This suggests that egg viability was lower on A. cruentus than on S. macrocarpon. Indeed many eggs of T. ludeni on A. cruentus failed to hatch  confirm these suggestions, as it was higher on S. macrocarpon than on A. cruentus. However, the data found in this study are lower than the 0.273 observed on cotton at 28°C by da Silva  was higher on to hatch , and juvrocarpon . The tem species . With S.da Silva , but fal species .A. cruentus may be due to its chemical composition as it may contain some so-called digestibility reducers known to exert sublethal effects by impairing herbivores' growth. Several studies have shown that population density and fecundity of various tetranychid mites on various host plants depend on plant quality (mr) of T. urticae on bean leaves and the potassium content of the leaves (see review by A. cruentus shows that its potassium content is much higher than that of S. macrocarpon (mr of T. ludeni on A. cruentus. Furthermore, some secondary plant metabolites such as phenolic compounds are also known to adversely affect pathogens and insect/mite populations (T. ludeni on A. cruentus compared with S. macrocarpon.The high mortality recorded during the immature stages on  quality . Some ch quality . Nitrogerocarpon , and thiulations , and thiT. ludeni used in these experiments came from S. aethiopicum, a solanaceous species close to S. macrocarpon. These mite populations might have, therefore, developed an adaptation to solanaceous species and this could enhance their performances on S. macrocarpon compared with A. cruentus, an Amaranthaceae. The higher net reproductive rate and the shorter doubling time of T. ludeni on S. macrocarpon may explain the relatively higher densities of T. ludeni generally observed on solanaceous plants in the field (The population of he field . It is whe field .T. ludeni are dependent on host-plant quality. Demographic parameters of this mite pest are better on S. macrocarpon than on A. cruentus, suggesting that S. macrocarpon is more suitable to T. ludeni than A. cruentus. This differential suitability of host plants to the mite is an important factor to consider while exploring IPM solutions for T. ludeni. For example, intercropping and/or rotation of A. cruentus and S. macrocarpon could be used to break the dissemination of T. ludeni within a vegetable farm. Moreover, a full knowledge of the chemical composition of potential host plants and/or non-host plants of the mite pest, and of how this can affect the growth of the mite might be an important key for developing biopesticides for a cheaper and environmentally safer control of mite pest on vegetable farms in Africa.This study shows that population density and fecundity of


Arvid Carlsson was born in Uppsala, Sweden in 1923. Dr. Carlsson, a pharmacologist, is best known for his contributions on the neurotransmitter, dopamine, for which he won the Nobel Prize in 2000 for Medicine/Physiology. The co-recipients were Dr. Eric Kendel and Dr. Paul Greengard.Dr. Carlsson entered Medical School in 1941 and his education was interrupted by several years of service in the Swedish armed forces. In 1951, he finished the M.L. degree, now equivalent to M.D. in North America. Later, he became a Professor at the University of Lund. In 1959, he moved to Goteborg University.In 1957, Dr. Carlsson showed that dopamine was a neurotransmitter in the brain and not just a precursor of norepinephrine.3 He did Dr. Carlsson was also instrumental in developing the ‘dopamine theory of schizophrenia’5 and theDr. Carlsson was also among the first researchers of the antidepressant compound, zimeldine, which was the first selective serotonin re-uptake inhibitor. The precursor of this drug was brompheniramine. Here, one should note that he did substantial work on the synthesis and metabolism of 5-hydroxytryptamine (serotonin) in the central nervous system. However,In the aging brain, there is a reduction of the levels of several transmitter substances and of the activities of enzymes involved in their synthesis and/or catabolism. CarlssonIt is interesting to note that Dr. Carlsson opposed fluoridation of water as it is a violation of modern pharmacological principles, and he succeeded in his campaign along with hundreds of other scientists. It is also important to note that the incidence of dental caries was the same in Sweden as compared to fluoridated countries such as the USA. As British Columbia in Canada was considering fluoridation of water, Carlsson said, ‘I would advise against fluoridation. He reiterated that individual prophylaxis (treatment) is preferable on principle grounds and that it is equally effective’.


Anopheles gambiae s.s mosquitoes are important vectors of lymphatic filariasis (LF) and malaria in Ghana. To better understand their ecological aspects and influence on disease transmission, we examined the spatial distribution of the An. gambiae (M and S) molecular forms and associated environmental factors, and determined their relationship with disease prevalence. Published and current data available on the An. gambiae species in Ghana were collected in a database for analysis, and the study sites were georeferenced and mapped. Using the An. gambiae s.s sites, environmental data were derived from climate, vegetation and remote-sensed satellite sources, and disease prevalence data from existing LF and malaria maps in the literature. The data showed that An. gambiae M and S forms were sympatric in most locations. However, the S form predominated in the central region, while the M form predominated in the northern and coastal savanna regions. Bivariate and multiple regression analyses identified temperature as a key factor distinguishing their distributions. An. gambiae M was significantly correlated with LF, and 2.5 to 3 times more prevalent in the high LF zone than low to medium zones. There were no significant associations between high prevalence An. gambiae s.s locations and malaria. The distribution of the An. gambiae M and S forms and the diseases they transmit in Ghana appear to be distinct, driven by different environmental factors. This study provides useful baseline information for disease control, and future work on the An. gambiae s.s in Ghana. Anopheles gambiae sensu lato (s.l.) contains seven species, of which An. gambiae sensu stricto (s.s), An. arabiensis and An. melas are three of the major vectors of lymphatic filariasis (LF) and malaria caused by Wuchereria bancrofti, and Plasmodium falciparum respectively in West Africa An. gambiae s.l and the An. funestus to be the major vectors of LF in the southern coastal zone and in the northern region of the country The mosquito Anopheles vectors prioritizes the use of insecticide treated materials and indoor residual spraying. However, the efficiency of these interventions will depend on information on the distribution and abundance of the main vectors, the specific molecular forms of An. gambiae s.s and the levels of insecticide resistance within them Vector control is considered an important tool for diseases transmitted by mosquitoes and other insect vectors An. gambiae complex in Africa An. gambiae s.s covered 11 sites across different ecological zones of the country An. gambiae S form predominated across the country, except in the arid north where only the M form was found. While this was the first major attempt at presenting the geographical distribution and ecological variations in An. gambiae s.s in Ghana, it is not known how these data compare with other An. gambiae s.s data, whether there are specific environmental factors driving the distributions of the M and S molecular forms, and if there are any associations with LF and malaria distributions Studies on the distribution of the An. gambiae s.s in Ghana, and aimed to i) examine the spatial distribution of the An. gambiae M and S forms across the country, ii) identify key environmental factors associated with their distribution, and iii) determine their relationship with LF and malaria prevalence distributions. It is envisaged that this information will help develop a comprehensive profile on the ecology of An. gambiae s.s in Ghana, which will assist researchers and the diseases control programs in the country.In order to better understand the ecological aspects of this important vector, their influence on the epidemiology of LF and malaria, this study collated data on The Republic of Ghana is a developing West African country bordered to the north by Burkina Faso, to the east by Togo, to the West by Cote d'Ivoire and to the south by the Gulf of Guinea An. gambiae s.l in Ghana were identified from various sources including published articles in peer-reviewed journals, unpublished works from MPhil and PhD theses held at the Noguchi Memorial Institute for Medical Research (NMIMR), Accra-Ghana, as well as on-going studies at NMIMR. The collated data spanned from 2001 to 2008. Information on the location, study period, sample size, collection method, mosquito species and molecular forms, from each study were collated into a database. Various collection methods including; human landing catches, pyrethroid spray catches, larval collections and aspirators were used, depending on the location of the study sites and the objectives of the various studies. However, irrespective of the collection method, data collected from the same location in different years were considered separately. With the exception of few sites, most locations (i.e collection site) in the database were geo-referenced using the latitude and longitude coordinates obtained by cross-checking the names with data from the GEOnet Names Server Entomological studies on An. gambiae s.l, and the different prevalence distributions of An. gambiae M and S across the country were mapped. Mosquito collection methods were also compared, to highlight differences in sampling. Second, spatial analysis of the An. gambiae M and S prevalences were examined using ArcGIS Spatial Analyst and Statistics tools . The Moran's I statistic was used to determine spatial autocorrelation patterns i.e. clustered, dispersed, random, and the Getis-Ord Gi* statistic was used to identify the specific locations where high and low prevalences were clustered  +1.96 and −1.96 standard deviations). In addition, the kernel density estimation (KDE) method, non-parametric way of estimating the probability density function, was used to create a continuous surface representing the high to low density distributions of each molecular form.All data were imported into the geographical information systems software ArcGIS 9.2  for mapping and spatial analyses. First, the overall distribution of the An. gambiae M and S in Ghana, specific data on elevation, vegetation, precipitation, temperature and humidity were obtained for each location (i.e. collection site), and compiled into a database, for descriptive and statistical analyses in SPSS 16.0 .To examine environmental factors associated with the prevalence distributions of Elevation data were derived from the U.S Geological Survey's ETOPO2 Digital Elevation Model available from ESRI . The elevation at each collection site was determined by importing the digital elevation map into ArcGIS 9.2 and extracting the underlying value (metres). Vegetation and climate data were based on the mean annual values for the specific year of study, obtained from the best available sources via the Climate Data Library of the International Research Institute for Climate and Society An. gambiae M and S, and each environmental variable was examined using bivariate correlations, Pearson's correlation coefficient . Stepwise multiple linear regression analysis was then used to identify the environmental factor that would best predict the distribution of each molecular form. To account for environmental variables that may be highly correlated with each other, the level of colinearity tolerance in the stepwise regression procedure was set at ≥0.8 and only variables above this threshold were accepted in the models. Second, to better understand the environmental parameters associated with the An. gambiae M and S forms, mean environmental measures between high and low prevalence sites were compared using the Mann-Whitney U test with Bonferroni correction for multiple comparisons.First, the relationship between An. gambiae M and S prevalence and the distribution of disease, maps on the LF prevalence P. falciparum prevalence W. bancrofti seroprevalence data collected in 2000 from 401 villages throughout Benin, Burkina Faso, Ghana and Togo P. falciparum malaria prevalence map was modelled on extensive data obtained from children aged 2–10 years in non-epidemic periods, using a generalized linear mixed model To examine the relationship between the An. gambiae s.s collection sites were used as focal points, whereby the underlying disease prevalence data could be compared with the entomological data. The LF and malaria prevalence data, corresponding to the latitude and longitude of each mosquito collection site, were extracted and exported for descriptive and statistical analyses, which included bivariate correlations, and comparison of means between high and low prevalence sites of the An. gambiae M and S forms.The Further, it was of particular interest to explore the entomological and environmental characteristics in different LF transmission zones based on the prevalence data map in Gyapong et al. 2002 An. gambiae species complex database contained 143 records with a total of 12,607 mosquitoes, reflecting both larval and adult catches. From this, the distribution of An. gambiae s.l was mapped , followed by An. melas  and An. arabiensis . The map indicates that the distribution of An. gambiae s.s was widespread, while An. melas was primarily found along the coastal Savanna zones, a predominantly marshy environment, and An. arabiensis mainly in the northern savanna zone. Anopheles funestus, the second most important vector of malaria and lymphatic filariasis in Ghana, was also recorded in 9 sites with a total of 1,825 mosquitoes. Refer to supplementary data File S1 “Anopheles distribution records in Ghana” for further information on An. funestus and other Anopheles mosquito species, collection sites and coordinates, year and month of collection (where available), collection methods, numbers collected and identified, and the data sources.The collated s mapped . The mosAn. gambiae s.s data, a total of 70 collection sites had information on the M form  and S form . The collection sites were located predominately in three geographical regions i.e. the south eastern, central western and north central. The distribution of An. gambiae M and S forms varied across the country in different proportions .Of the portions . Figure An. gambiae M  and An. gambiae S  forms. The resultant Z scores of the Getis-Ord Gi* hot spot analyses (using inverse-distance weighting), indicated similar trends with significantly different clustering of high and low prevalences of the An. gambiae M and S forms. These spatial trends were overlaid a density distribution surface map shown in While, it is possible that the geographical grouping of collection sites may influence trends, the spatial analyses carried out in this study indicated a positive spatial autocorrelation or clustering for both the An. gambiae M and S forms, and the environmental variables are shown in An. gambiae M form was significantly positively associated with temperature (r = 0.51), and negatively with elevation (r = −0.28), precipitation (r = −0.33), and humidity (r = −0.26). This contrasts to the An. gambiae S form, which was found to be significantly negatively associated with temperature (r = −0.58), and positively with elevation (r = 0.30) and rainfall (r = 0.41). Interestingly, elevation, precipitation and temperature correlations increased when data were stratified by the two main collection methods, HLC and larval collections  and for An. gambiae S, 36%  of the variance in the model  between elevation, precipitation and temperature for both An. gambiae M and S forms between the high and low prevalence areas.For each molecular form, comparisons of environmental measures between locations with significantly high and low prevalences, defined by positive Z scores (≥+1.96) and negative Z scores (≤−1.96) respectively, are shown in An. gambiae M and S forms was plotted across a range of environmental groupings , while An. gambiae S form was significantly negatively associated with LF (r = −0.48), but positively with malaria (r = 0.26). Interestingly, correlation analysis between each mosquito species, and each disease indicated significant negative associations between An. gambiae M and An. gambiae S (r = −0.76), and between LF and malaria prevalence (r = −0.41).The relationship between the An. gambiae M and S sites (as described above). Overall, locations with high An. gambiae M prevalences (Z scores≥+1.96), were found to have significantly higher LF prevalences (20%) than those locations with low prevalences (Z scores≤−1.96), by these species (2.4%) and/or where the prevalence of An. gambiae S form was significantly high (2.2%). No significant differences were found between malaria prevalence and each mosquito species  and temperatures (26.8°C) were found to be significantly higher (±2SE), and An. gambiae S prevalence (25.6%) significantly lower, than those found in medium transmission  and low transmission  zones. In the zero/low transmission zone, precipitation measures (2.83 mm) were found to be significantly higher (+2SE), than those in the medium transmission (2.34 mm) to high transmission (1.99 mm) zones  zones . Refer tAn. gambiae s.l in Ghana et al. attempted a broader coverage, this study represents the first nationwide review aimed at identifying the distribution of the various member species of the An. gambiae s.l, the molecular forms of the An. gambiae s.s, together with key environmental drivers and how they may relate to diseases. This study confirms previous observations that An. gambiae s.l is the major human biting mosquito species in Ghana An. gambiae s.l, An. gambiae s.s is the predominant species An. gambiae s.l found in Ghana are An. arabiensis and An. melas.Most entomological studies on An. gambiae M and S forms varied significantly across the country. The two molecular forms were found sympatrically in most locations, except in some areas in the middle region of the country where only the An. gambiae S form was observed, and in certain areas in the northern savanna and coastal savanna areas where only the An. gambiae M form was observed. This is confirmed by the spatial analysis and high Z score values in An. gambiae M/S form dominant areas. Also, the clustering remained relatively consistent irrespective of the different collection methods. Along the coast and in the northern savanna, the An. gambiae M form was predominant and clustered, while the S molecular form was most common and clustered in the middle belt. This positive spatial autocorrelation indicates that An. gambiae M/S distributions are geographically defined, and nearby areas are likely to comprise the same or similar species compositions, than those further away. The distribution of each species is also influenced by distinct and geographical related environmental factors and habitat characteristics. For example, the dominance of the An. gambiae M form in the northern and coastal savanna areas may be due to the wide presence of permanent breeding conditions provided by irrigation facilities An. gambiae S form in the middle region of the country may be explained by the fact that this region is mountainous, forested, with lower mean temperatures and the highest recorded rainfall in Ghana, which supports the findings of our study. In sub-Saharan Africa the abundance and distribution of Anopheles mosquito species is dependent on environmental factors and ecological zones An. gambiae M and S forms, which appear to be driven by a range of environmental factors. The relation between human population density and vector distribution is an indirect one and difficult to measure. However, it may be inferred that the NDVI, which is a measure of vegetation greenness and density, is somehow affected by the degree of land occupancy and exploitation by humans through the action of occupation and clearance for agricultural and developmental purposes.The distribution of the An. gambiae S form in a larger part of the country from this study confirm suggestions that the An. gambiae S form, has broader environmental ranges, and therefore is found in more locations than the M form An. arabiensis and An. gambiae s.sAn. gambiae M form was more prevalent and seemingly better adapted to higher temperatures, than the S form. This is in agreement with the suggestion that the An. gambiae M form shows a more latitudinal range in West Africa than the S form Variations in the vector population densities of these two molecular forms have been observed in populations in Mali and Cameroon as well as between the various chromosomal forms in Mali An. gambiae S form, whilst high LF prevalence and high transmission zones were associated with high temperatures and significantly high An. gambiae M prevalences. This relationship between An. gambiae M and S forms with LF and malaria supports previous studies that suggest the Mopti form of An. gambiae s.s is more associated with W. bancrofti than malaria transmission An. gambiaeAn. gambiae Mopti chromosomal form dominant areas The mosquito vectors' association with each disease might explain the negative association between LF and malaria described by Kelly-Hope and colleagues An. gambiae M and S forms in Ghana, highlighting the environmental factors that may play a role in determining their distributions. The information provided also marks a beginning in understanding the LF disease distribution pattern in Ghana relative to the forms of the An. gambiae s.s. Despite the limitations of this study in using previously modeled LF and malaria data, these results are very useful for disease control and allocation of resources, especially for LF, which together with its vectors appear to be restricted to hotter, less elevated regions of the country. Another limitation to this study is the small number of sites involved in the analysis of the An. gambiae M and S distribution. This coupled with their uneven distribution between the southern half and the northern half of the country could introduce potential bias in the analyses. Undertaking spatial statistical analysis with a larger dataset in the future, and further modeling their distributions through ecologic niche modeling The results from this study provide useful information on the distribution of the File S1Data file for Anopheles mosquito species, collection sites and coordinates, year and month of collection (where available), collection methods, numbers collected and identified, and the data sources.(0.04 MB XLS)Click here for additional data file.File S2Summary of entomological and environmental variables in different LF transmission zones.(0.08 MB DOC)Click here for additional data file.


In biopsies from the oral mucosa of 235 cases in which the diagnosis was lichen planus, keratosis or leukoplakia, mitotic values were calculated for the stratum basale  and the stratum spinosum (M.V. spinous). The mean M.V. basal was significantly different from the mean M.V. spinous in the keratosis and leukoplakia groups, but not in the lichen planus group. Within the keratosis and leukoplakia groups, M.V. basal and M.V. spinous were significantly correlated. When each of the mean M.V.s was compared with the M.V.s for the other diagnostic groups, various significant differences were found. The M.V.s were examined in relation to the type of keratinization, the presence of acanthosis or atrophy, and the patient's age, but the M.V.s were not significantly related to these features.


The Magpie Trial, a randomised trial comparing magnesium sulphate with placebo for women with pre-eclampsia. This paper describes methods used for follow up in the UK, and presents additional data collected.In the UK 774 women and their 827 children were included; excluded were women discharged without a surviving child and families who opted out. General practitioners were sent a questionnaire when the child was around 18 months old. When the child was two years, or older, questionnaires asking about the health of the women and children were posted to families. A sample of families was offered a home visit, during which the child was assessed using the Bayley Scales of Infant Development.Of the women, 12 were lost to follow up and three died. Of the children, 12 were lost to follow up, 5 were excluded and 19 died. General practitioners returned 688/759 (91%) questionnaires, as did 619/759 (82%) women. Responses were largely comparable. 32 women had serious morbidity potentially related to pre-eclampsia. 30% of children were reported to have been admitted to hospital. There were no clear differences between the randomised groups in the child's behaviour, women's fertility or use of health service resources.Data presented here provide further reassurance about the longer term safety of magnesium sulphate when used for women with pre-eclampsia. Postal questionnaires in the UK to assess the longer term health and wellbeing of women and children recruited to trials are feasible, and can achieve a high response rate. Responses from families and general practitioners were comparableTrial registration number of the Magpie Trial [ISRCTN86938761] Pre-eclampsia; a multisystem disorder of pregnancy usually associated with raised blood pressure and proteinuria, complicates 2–8% of pregnancies. Althougin utero exposure to magnesium sulphate is associated with an increase in the risk of death or neurosensory disability for the child [The Magpie Trial compared magnesium sulphate with placebo for women with pre-eclampsia. This rahe child  , or in he child .For the follow up study; methods for tracing and assessing families were developed that could be implemented across 19 countries and a range of settings.,12 NeverBetween July 1998 and November 2001, 804 women were recruited to the Magpie Trial at 67 UK hospitals. Follow up was from October 2002 until May 2004. Thirty women were excluded from tracing leaving 774 eligible for follow up  with som1) Does your child play happily with toys for up to 10 minutes?2) Is your child frightened in new situations?3) Does your child play happily with other children if you are around?4) Does your child cling to you when you are with other people?5) Does your child settle easily to sleep throughout the night?Options for response to questions 'Yes' 'Sometimes' 'No'6) Has your child been prescribed any medicines to be taken for more than two weeks?If yes, please give their names ..................................................................7) Over the last 3 – 6 months have you taken your child to the general practitioner?If yes, how many times and what for ............................................................The accompanying women's questionnaire asked women about their own health, again with additional questions used only in the UK:1) Have you used contraception since your child was born?If yes which ones ................................................................2) Did having pre-eclampsia (toxaemia) contribute to your choice?If yes, please explain how ...........................................................3) Have you tried to get pregnant again since your child was born?If no, did having pre-eclampsia contribute to your decision?Please explain ...................................................................4) Have you seen anyone else about your health, or how you feel, since your child was born?If yes, who and what for? .......................................Options for response to questions 'Yes' or 'No'Women were also asked about their experience of participating in the Magpie Trial. Responses to these questions are part of a qualitative analysis of participants' views of being in the trial, and are reported elsewhere .Children were considered 'screen positive' if they failed the ASQ, if the ASQ could not be scored, or if the child passed an ASQ for a younger age group. The families of these children were contacted by telephone and offered a home visit for further assessment. Children were considered screen negative if they passed the ASQ for their own age group, or for an older child. Screen negative children in the north east of England were offered a home visit.The aim of the home visit was to confirm whether or not the child had neurodevelopmental delay, or any other significant problem. If so, to collect information that would, if possible, establish a diagnosis. Families were asked about the child's current health and development, and the child was tested using the Bayley Scales of Infant Development (BSID-II) . These aDuring the home visit we took the opportunity to measure the woman's blood pressure, and to ask her about her health. Home visits lasted approximately two to three hours. All assessments were blind to the allocated treatment.Responses by the general practitioner and the family were described and compared for both women and children. As information was collected from general practitioners earlier than from families, the aim was to assess whether these responses were comparable and whether differences could reasonably be expected to be due to the time difference. Data assessing the child's behaviour, the woman's fertility, and their use of health service resources were compared between the two allocated treatment groups.Of the 774 UK women included in follow up, 12 were lost to follow up and three died after discharge from hospital  after the child was born (adjusted for gestation at birth up to 24 months). There was no difference in the response rate between the two allocated groups. In addition, there were no clear differences between women who responded to the questionnaire (n = 619) and those who did not (n = 140) in characteristics at trial entry, outcome at delivery or outcome at discharge following delivery (data not shown).Of women included in the UK follow up, three quarters (74%) were primiparous, a quarter were less than 34 weeks gestation at trial entry, just over half (56%) had labour induced, two thirds (61%) were delivered by caesarean section and nearly half (46%) had severe pre-eclampsia (data not shown). For 132 children the family did not complete the questionnaire; but for two the families contacted us to say the child had severe disability and request that they have no further assessment. The remaining 130 children were no more likely to have developmental delay, reported by the general practitioner, than those children for whom the family completed a questionnaire (data not shown).Overall for 688/759 (91%) women the general practitioner's questionnaire was completed. The majority (62%) responded after the initial questionnaire, some practices requiring reminders (34%). Fifty-one practices requested, and were sent, a copy of the trial consent form; of these 36 responded by returning the completed questionnaire. Payment was requested and provided for 36 practices . Five practices gave no response. No general practitioner could be identified for three women. The general practitioner's questionnaires were completed 23 months  after the child was born (adjusted for gestation at birth up to 24 months). Based on information provided by the general practitioner we did not contact four families: the reasons were recent marital breakdown (2), recent death of husband (1) and drug addiction (1).Questionnaire responses were available from both general practitioners and women for 551/759 women Table . For theOverall serious morbidity potentially related to pre-eclampsia was reported for 31 (4%) women recruited in the UK; of whom 26 had severe hypertension (defined as taking two or more antihypertensives at time of completing questionnaire), six had renal problems, and one woman had had a stroke. Other morbidity was reported for 331 (44%) women, this included: minor ailments  for 235 (31%) women, major psychiatric illness (psychosis or depression requiring treatment) for 144 (19%) and minor psychiatric illness (stress or anxiety not requiring treatment) for 49 (6%). Women's reporting of their fertility since the Magpie Trial and family history of pre-eclampsia and use of health care resources are in Table Over 30% of children were reported to have been admitted to hospital: chest infection and minor surgery being the most common reasons. Other morbidity was reported for 149 26%) children, this included: minor ailments . Prescribed medication for more than two weeks was reported for 126 (25%) children: these included vitamins (29%), asthma treatment (25%) and antibiotics (19%)  receptors in the brain, the pathway for anoxic cell damage, we hypothesized that al delay . These aThe high response rate to the postal questionnaires is a key strength of our study. By sending questionnaires to both the families and general practitioners we were able to achieve 98% follow up. For families we were unable to contact, or who did not respond to the questionnaire, information was provided by their general practitioner. Comparison of these responses is reassuring, in that they are largely comparable. This is consistent with the agreement between a parental questionnaire and paediatric assessment for children with a high prevalence of impairment . The maiThe follow up study did not begin until after recruitment closed, as this was when funding was secured. Nevertheless, use of the Office for National Statistics ensuring complete ascertainment of mortality, and accessing the National Health Service Tracing Scheme enabled us to trace all but four surviving mothers and their four children. An additional advantage of the separate system for ascertaining deaths was that unnecessary distress of contacting families if the child had died could be avoided, and if the mother died an appropriate approach could be made to the child's current carer.Fewer families responded to the questionnaires than did the general practitioners. Our children's questionnaire asked for the child's current height and weight, information that was often missing from questionnaires returned. Some families told us they delayed returning questionnaires in order to take the child to a clinic to be weighed and measured. It is seems likely that this delay contributed to the non-response. As the data for height and weight were incomplete, they have not been reported. Our impression is that women and families largely valued being contacted for the follow up study. Only 12 families chose to opt out. Women returning the questionnaires, and being contacted for a home visit, often expressed their appreciation of our continued interest in their, and their child's, health and welfare. In addition, the questionnaire to women included three questions about their views of participation in the Magpie Trial, these data are reported elsewhere.Data presented here provide further confirmation of the considerable morbidity experienced by women following childbirth. There is a growing awareness of the high level of health problems women experience, even after an uncomplicated pregnancy and childbirth . Around Magnesium sulphate reduces the risk of eclampsia  without The authors declare that they have no competing interests.RMDS wrote the first draft of the manuscript, with input from LD. All authors participated in reviewing and revising the manuscript and approved its final version.The pre-publication history for this paper can be accessed here: