A comparative parasitologic study on Biomphalaria glabrata snail and C3H/He mice infected with human and murine isolates of Schistosoma mansoni derived from Sumidouro, Rio de Janeiro, Brazil

Experiments were carried out to analyze the biological characteristics of two sympatric isolates of Schistosoma mansoni derived from humans and murines in a low endemic transmission area (Sumidouro county, state of Rio de Janeiro, Brazil). Sympatric reared-laboratory Biomphalaria glabrata and C3H/He mice were used as experimental hosts. Parameters assessed comprised: precercarial period, infectivity and mortality (snails), prepatent period, infectivity (percentage of cercariae maturation into adult worm) and intestinal egg count (mice). The murine isolate showed a shorter precercarial period and higher infectivity than human isolate (p < 0.05). This biological heterogenicity did not correspond to the vertebrate data because any biological parameter presented significant difference (p > 0.05). These data suggest that both isolates are local sub-populations, providing support for the hypotheses that in a same biotope mixed populations or sub-populations circulate among their main host (human beings) and/or rodent as an anfixenous infection.

The role of urea in the oviposition behaviour of Japanese encephalitis vectors in rice fields of South India

The effect of urea on the oviposition behaviour of culicine vectors of Japanese encephalitis was studied in rice fields. Gravid females had a strong preference for oviposition in urea treated areas in rice fields, while no such preference was exhibited in untreated areas. The egg laying declined in the area where urea treated water surface had a mechanical barrier, which allowed volatile fractions to escape, but prevented contact with the water. Urea was shown to act as an oviposition attractant/stimulant for Culex tritaeniorhynchus, but its role was not clear for Cx. vishnui, as the number of egg rafts obtained for the latter species was low.

Host nutritional status as a contributory factor to the remodeling of schistosomal hepatic fibrosis

Weaning Swiss mice were percutaneously infected with 30 cercariae of Schistosoma mansoni and submitted to a shifting either from a deficient to a balanced diet or vice-versa, for 24 weeks. The nutritional status was weekly evaluated by measurements of growth curves and food intake. Hepatic fibrosis and periovular granulomas were studied by histological, morphometric and biochemical methods. All mice fed on a deficient diet failed to develop periportal "pipestem" fibrosis after chronic infection. An unexpected finding was the absence of pipestem fibrosis in mice on normal diet, probably related to the sample size. The lower values for nutritional parameters were mainly due to the deficient diet, rather than to infection. Liver/body weight ratio was higher in "early undernutrition" group, after shifting to the balanced diet. Volume density and numerical density of egg granulomas reached lowest values in undernourished animals. The amount of collagen was reduced in undernourished mice, attaining higher concentrations in well-fed controls and in "late undernutrition" (balanced diet shifted to a deficient one), where collagen deposition appeared increased in granulomas. That finding suggested interference with collagen degradation and resorption in "late" undernourished animals. Thus, host nutritional status plays a role in connective tissue changes of hepatic schistosomiasis in mice.

Detection of parasite eggs from archaeological excavations in the Republic of Korea

Excavations at two sites dating from 2000 BC-1900 AD in southeastern areas of the Republic of Korea, revealed the remains of several structures. Examination of the contents suspected privies revealed the presence of eggs from 5 kinds of parasite: Ascaris, Trichuris, Clonorchis, and two species of unknown trematodes. Clonorchis sinensis eggs were found in a soil dating from around AD 668-935. This is the first record of C. sinensis eggs in archaeological materials in the Republic of Korea.

Phenotypic plasticity in adult worms of Schistosoma mansoni (Trematoda:Schistosomatidae) evidenced by brightfield and confocal laser scanning microscopies

A comparative morphometric study was performed to identify host-induced morphological alterations in Schistosoma mansoni adult worms. A wild parasite population was obtained from a naturally infected rodent (Nectomys squamipes)and then recovered from laboratory infected C3H/He mice. Furthermore, allopatric worm populations maintained for long-term under laboratory conditions in Swiss Webster mice were passed on to N. squamipes. Suckers and genital system (testicular lobes, uterine egg, and egg spine) were analyzed by a digital system for image analysis. Confocal laser scanning microscopy (CLSM) showed details of the genital system (testicular lobes, vitelline glands, and ovary) and the tegument just below the ventral sucker. Significant morphological changes (p < 0.05) were detected in male worms in all experimental conditions, with no significant variability as assessed by CLSM. Significant changes (p < 0.05) were evident in females from the wild population related to their ovaries and vitelline glands, whereas allopatric females presented differences only in this last character. We conclude that S. mansoni worms present the phenotypic plasticity induced by modifications in the parasite's microenvironment, mainly during the first passage under laboratory conditions.

A review of coccidiostat residues in poultry

safefood, the Food Safety Promotion Board, is responsible for increasing food safety awareness and for supporting north/south scientific co-operation. safefood is currently funding a project entitled "Poultry Meat: improving food safety by improving chemical residue surveillance". This joint project between the Veterinary Sciences Division, Queen's University, Belfast and the National Food Centre, Teagasc, Dublin, is addressing the problem of anti-coccidial drug residues in poultry meat and eggs through an all-island research and residue testing initiative. The project started in 2001 and will continue until 2004. Poultry have a high susceptibility to the parasitic disease, coccidiosis. Because of this susceptibility, veterinary drugs, commonly known as coccidiostats are routinely used in intensively-reared poultry. The coccidiostats are potent drugs and, where residues occur in food, they may exacerbate certain coronary disease conditions. It is important, therefore, for poultry and egg producers to prevent the occurrence of residues of coccidiostats in food products.

Seasonal pattern of abundance of Aedes aegypti (Diptera: Culicidae) in Buenos Aires city, Argentina

In Buenos Aires, the most crowded city of Argentina, there is a potential risk of dengue virus transmission by the mosquito Aedes aegypti during late summer. The temporal patterns of oviposition activity and abundance of breeding sites of this vector were studied in two cemeteries of the city. Between September 1998 and August 1999, we examined 142 ovitraps weekly and a total of 18,010 water-filled containers. Both study areas showed remarkable differences in the percentages of positive ovitraps (19% vs 8%) and breeding sites (18% vs 1%), but similar temporal abundance patterns. The percentage of breeding sites was higher in summer and autumn than in spring and winter, and the percentage of positive ovitraps was higher in summer than in the other three seasons. Immatures were recorded from the first week of October to the second week of July, and oviposition activity from the third week of October until the end of April. In both cemeteries and with both methodologies the highest infestation levels were registered in March (ovitraps: 41.8% and 20.6%, breeding sites: 39.2% and 3.4%). These highest abundances took place after several months with mean temperatures above 20ºC and accumulated rainfalls above 150 mm. A sharp decline in oviposition activity was observed when monthly mean temperature decreased to 16.5ºC, and no eggs were found below 14.8ºC. Seasonal fluctuation of Ae. aegypti abundances in mid-latitudes like Buenos Aires would allow reduction of the egg mosquito population through the elimination of containers during the coldest months, which are free of adults.

Life tables and reproductive parameters of Lutzomyia spinicrassa (Diptera: Psychodidae) under laboratory conditions

Lutzomyia spinicrassa is a vector of Leishmania braziliensis in Colombia. This sand fly has a broad geographical distribution in Colombia and Venezuela and it is found mainly in coffee plantations. Baseline biological growth data of L. spinicrassa were obtained under experimental laboratory conditions. The development time from egg to adult ranged from 59 to 121 days, with 12.74 weeks in average. Based on cohorts of 100 females, horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (8.4 females per cohort female), generation time (12.74 weeks), intrinsic rate of population increase (0.17), and finite rate of population increment (1.18). The reproductive value for each class age of the cohort females was calculated. Vertical life tables were elaborated and mortality was described for the generation obtained of the field cohort. In addition, for two successive generations, additive variance and heritability for fecundity were estimated.

Leishmania major-specific B cells are necessary for Th2 cell development and susceptibility to L. major LV39 in BALB/c mice.

B lymphocytes are considered to play a minimal role in host defense against Leishmania major. In this study, the contribution of B cells to susceptibility to infection with different strains of L. major was investigated in BALB/c mice lacking mature B cells due to the disruption of the IgM transmembrane domain (microMT). Whereas BALB/c microMT remained susceptible to infection with L. major IR173 and IR75, they were partially resistant to infection with L. major LV39. Adoptive transfer of naive B cells into BALB/c microMT mice before infection restored susceptibility to infection with L. major LV39, demonstrating a role for B cells in susceptibility to infection with this parasite. In contrast, adoptive transfer of B cells that express an IgM/IgD specific for hen egg lysozyme (HEL), an irrelevant Ag, did not restore disease progression in BALB/c microMT mice infected with L. major LV39. This finding was likely due to the inability of HEL Tg B cells to internalize and present Leishmania Ags to specific T cells. Furthermore, specific Ig did not contribute to disease progression as assessed by transfer of immune serum in BALB/c microMT mice. These data suggest that direct Ag presentation by specific B cells and not Ig effector functions is involved in susceptibility of BALB/c mice to infection with L. major LV39.

The initial epidemiological studies in the low endemicity schistosomiasis area in esteio, Rio Grande do Sul, the southernmost Brazilian State, 1997 to 2000

Nor Biomphalaria glabrata neither Schistosoma mansoni were reported from Rio Grande do Sul, the southernmost Brazilian state before 1997. Their detection next to the Sinos River, Esteio, confirmed predictions of schistosomiasis expansion to the south. Parasitological examinations both in snails and fecal samples from the human population were performed from 1997 to 2000. The last 3 out of 5 surveys were performed after a preliminar serological screening procedure in a risk group identified at a population census. A total of 11 infected individuals were found infected and snails from 2 different sites were positive for S. mansoni. Samples from these 2 and other sites were identified as B. glabrata. Egg counts in feces were below 1 per gram in 6 out of 11 patients. Some socio-cultural perceptions of water contact activities next to the Sinos River may cause difficulties to control efforts, but they also may be partially acting against a very rapid increase in transmission intensity. The southernmost schistomiasis mansoni foci in Americas rise the alert for its ongoing expansion.

Malnutrition and hepatic fibrosis in murine schistosomiasis

In this paper, four different approaches attempting to reproduce the schistosomal liver fibrosis in undernourished mice are reported: shifting from a deficient to a balanced diet and vice-versa, repeated infections, influence of the genetic background, and immunological response. Infections were performed with 30 cercariae of Schistosoma mansoni and lasted at least four months. Undernourished mice were unable to reproduce the picture of "pipestem" fibrosis, except the C57 BL/10 inbred strain, four out of 21 mice developing the liver lesion. A link of this histological finding to the type of parasite strain can not be discarded at the moment. Repeated infections increased collagen deposition mainly in well nourished animals (seven out of 16 Swiss mice developed "pipestem"-like fibrosis). In undernourished infected Swiss mice the serum levels of soluble egg antigen specific antibodies IgG1, IgG2a, IgG2b, and IgG3 were two to four times lower than those detected for well nourished controls. The decreased humoral immune response coupled to the morphological, morphometric, and biochemical results reinforce the influence of the host nutritional status on the connective tissue changes of hepatic schistosomiasis.

Sativa seeds against Schistosoma mansoni different stages

The schistosomicidal properties of Nigella sativaseeds were tested in vitro against Schistosoma mansoni miracidia, cercariae, and adult worms. Results indicate its strong biocidal effects against all stages of the parasite and also showed an inhibitory effect on egg-laying of adult female worms. In the present work we also studied the effects of crushed seeds on some antioxidant enzymes; which have a role in protection of adult worms against host oxidant killing; as well as some enzymes of glucose metabolism; which have a crucial role in the survival of adult worms inside their hosts. The data revealed that the used drug induce an oxidative stress against adult worms which indicated by a decrease in the activities of both antioxidant enzymes, superoxide dismutase, glutathione peroxidase, and glutathione reductase and enzymes of glucose metabolism, hexokinase and glucose-6-phosphate dehydrogenase. Disturbing of such enzymes of adult worms using N. sativa seeds could in turn render the parasite vulnerable to damage by the host and may play a role in the antischistosomal potency of the used drug.

Identification of the best ovitrap installation sites for gravid Aedes (Stegomyia) aegypti in residences in Mirassol, state of São Paulo, Brazil

This study aimed at identifying the best ovitrap installation sites for gravid Aedes aegypti in Mirassol, state of São Paulo, Brazil. Ovitraps were installed in ten houses per block over ten blocks. Four ovitraps were placed per residence, one in the bedroom, one in the living room, and two outdoors with one in a sheltered area and one in an outside site. Each week for eleven weeks, visits were made to examine the ovitraps and to change the paddles used for egg-laying. Eggs were analyzed according to the trap location. The results showed that the outdoor sites received significantly more oviposition than indoor sites. Additionally, in respect to the outdoor sites, the outside site received significantly more oviposition than the sheltered site. A b correlation was observed between positive traps and egg numbers. The results are discussed with respect to the best installation site of the traps and their implications in surveillance and control of dengue vectors.

Analysis of the regulation of Nodal function by SPC-mediated cleavage during early mammalian development

RÉSUMÉ Après implantation dans l'utérus, le foetus de mammifère est composé de trois populations différentes de cellules: l'epiblast, l'ectoderme extraembryonnaire et l'endoderme viscéral. Pendant la gastrulation, les cellules de l'epiblast donnent naissance aux trois lignées germinales: l'ectoderme, le mésoderme et l'endodermes. Les lignées germinales produisent par la suite les différents tissus et organes du corps embryonnaire et adulte. Les cellules de l'ectoderme extraembryonnaire donnent par la suite le composant foetal du placenta qui est essentiel à la survie de l'embryon dans l'utérus. L'épiblast et l'ectoderme extraembryonnaire sont entourés par l'endoderme viscéral et forment une structure connue sous le nom de bouton embryonnaire. L'endoderme viscéral joue un rôle important dans l'embryogenèse car il comporte une sous-population de cellules appelées l'endoderme viscéral antérieur dont les signaux influencent l'épiblast adjacent et déterminent le futur axe antéro-postérieur de l'embryon. La protéine de signalisation Nodal de la famille des TGFß est essentielle dans l'épiblast pour spécifier le mésendoderme, l'endoderme viscéral antérieur, ainsi que pour maintenir les cellules souche de l'ectoderme extraembryonnaire. Ainsi, dans les embryons mutants pour Nodal, aucun axe antéro-postérieur n'est établi, les lignées germinales ne sont pas spécifiés et le placenta ne se développe pas. Au niveau moléculaire, comme pour les protéines de la famille des TGFß, Nodal est initialement synthétisée sous forme de précurseur avant d'être clivée de façon endoproteolytique par des protéanes sécrétées, les proprotéines convertases du type subtilisin (SPC), qui suppriment la partie inhibitrice N-terminale du pro peptide. Dans ce contexte, le projet de ma thèse a été d'analyser l'influence des SPC sur la fonction de Nodal en employant une combinaison d'approches génétiques et biochimiques. Premièrement, nous avons constaté que le clivage du précurseur par les protéases active Nodal, mais en même temps augmente son turn-over et diminue la portée de son action. Deuxièmement, dans l'embryon, il apparaît que Nodal est activé par l'action combinée de Furin et de PACE4, deux protéases sécrétées qui sont spécifiquement exprimées dans les cellules de l'ectoderme extraembryonnaire, donc adjacentes au domaine d'expression de Nodal. De manière similaire aux mutants de Nodal, les embryons mutants pour les deux protéases ne forment pas d'endoderme viscéral antérieur et ne gastrulent pas. Cependant, certains gènes cible de Nodal restent exprimés, suggérant que toutes les activités de Nodal ne sont pas dépendent du clivage par les SPCs. En effet, la génération et l'analyse de mutants portant un allèle knock-in qui code pour une forme mutante de Nodal résistante aux SPC, ont montré que ces mutants ont les caractères phénotypique des mutants de Nodal seulement de façon partielle. La formation de mésoderme est partiellement induite, et de façon remarquable, la forme de Nodal résistante aux SPC est capable d'agir à une distance de sa source, maintenant l'expression de ses propres protéases et d'autres gènes essentiels pour la spécification de l'ectoderme extraembryonnaire. Ensemble, ces résultats prouvent que par leur action directe les protéases extraembryonnaire modulent la signalisation de Nodal pendant le développement mammifère précoce. SUMMARY : Early after implantation in the uterus, the mammalian conceptus is composed of three different cell populations: the epiblast, the extraembryonic ectoderm and the visceral endoderm. During gastrulation, epiblast cells give rise to the three embryonic germ layers: the ectoderm, the mesoderm and the endoderm. These germ layers then generate the different tissues and organs of the embryonic and adult bodies. In parallel, extraembryonic ectoderm cells give rise to the fetal component of the placenta, which is essential for the survival of the embryo in the uterus. Both the epiblast and extraembryonic ectoderm are surrounded by the visceral endoderm to form a structure known as the egg cylinder. The visceral endoderm plays an important role as it harbours a subpopulation of cells called the anterior visceral endoderm, from which signals influence the adjacent epiblast and determine the future antero-posterior embryonic axis. The TGFß-related signalling protein Nodal is required within the epiblast to specify the mesoderm, the endoderm,the anterior visceral endoderm and is also essential to maintain stem cells in the extraembryonic ectoderm. Thus, in Nodal null conceptuses, no antero-posterior axis is established, the germ layers are not specified and the placenta does not develop. At the molecular level, Nodal, like related proteins of the TGFß family, is initially synthesized as a precursor and undergoes endoproteolytic cleavage by secreted proteases of the subtilisin-like proprotein convertases (SPC) to remove an inhibitory N-terminal pro peptide. In the embryo, Nodal is activated by the combined action of Furin and PACE4, two secreted SPCs that are specifically expressed in cells of the extraembryonic ectoderm, thus adjacent to the Nodal expression domain. Similar to Nodal null .embryos, mutant embryos lacking both these proteases fail to specify the anterior visceral endoderm and to undergo gastrulation. However, these mutants still express a subset of Nodal target genes, suggesting that part of Nodal activity is independent on cleavage by SPCs. Indeed, by generating and analyzing mutants with a knock-in allele that encodes an SPC-resistant mutant form of Nodal, I could show that they retain a subset of Nodal activities. Mesoderm formation is partially induced, but most remarkably, SPC-resistant Nodal form is able to act at a distance from its source, maintaining the expression of its proteases and of other genes essential for maintenance of the extraembryonic ectoderm.

Pre-hatching maternal effects and the tasty chick hypothesis

Question: Are maternal effects (i.e. maternal transfer of immune components to their offspring via the placenta or the egg) specifically directed to the offspring on which ectoparasites predictably aggregate? Organisms: The barn owl (Tyto alba) because late-hatched offspring are the main target of the ectoparasitic fly Carnus hemapterus. Hypothesis: Pre-hatching maternal effects enhance parasite resistance of late- compared with early-hatched nestlings. Search method: To disentangle the effect of natal from rearing ranks on parasite intensity, we exchanged hatchlings between nests to allocate early- and late-hatched hatchlings randomly in the within-brood age hierarchy. Result: After controlling for rearing ranks, cross-fostered late-hatched nestlings were less parasitized but lighter than cross-fostered early-hatched nestlings. Conclusion: Pre-hatching maternal effects increase parasite resistance of late-hatched offspring at a growth cost.