Experimental heteroxenous cycle of Lagochilascaris minor Leiper, 1909 (Nematoda: Ascarididae) in white mice and in cats

Reports of natural infections of sylvatic carnivores by adult worms of species similar to Lagochilascaris minor in the Neotropical region led to attempts to estabilish experimental cycles in laboratory mice and in cats. Also, larval development was seen in the skeletal muscle of an agouti (Dasyprocta leporina) infected per os with incubated eggs of the parasite obtained from a human case. In cats, adult worms develop and fertile eggs are expelled in the feces: in mice, larval stages of the parasite develop, and are encapsulate in the skeletal muscle, and in the adipose and subcutaneous connective tissue. From our observations, we conclude that the larva infective for the mouse is the early 3rd stage, while for the final host the infective form is the later 3rd stage. A single moult was seen in the mouse, giving rise to a small population of 4th stage larvae, long after the initial infection.

Pterigodermatites (Paucipectines) spinicaudatis n.sp. (Nematoda: Rictularidae) from Dromiciops australis (Marsupialia: Microbiotheriidae) in Bariloche, Rio Negro, Argentina, biogeographical distribution and host-parasite relationships

Pterigodermatites (P.) spinicaudatis sp.n. from Dromiciops australis is proposed and described. The simple morphology of the ovijector and the presence of a well developed spine between the two cuticular projections at the caudal extremity of the female distinguish the studied nematode from the remainder species of the genus parasitizing South American Edentata, marsupials and cricetid rodents. The distribution area of the hosts of the different species of P. (P.) are given. The studied genus does not parasitize any Australian marsupials. It was found in the endemic South American Microbiotheriidae. This fact suggests from a parasitological point of view that D. australis is not related to the Australian marsupials but to the South American ones.

Use of glass beads and CF 11 cellulose for removal of leukocytes from malaria-infected human blood in field settings

Passage of malaria infected blood through a two-layered column composed of acid-washed glass beads and CF 11 cellulose removes white cells from parasitized blood. However, because use of glass beads and CF 11 cellulose requires filtration of infected blood separately through these two resins and the addition of ADP, the procedure is time-consuming and may be inapropriate for use in the field, especially when large numbers of blood samples are to be treated. Our modification of this process yields parasitized cells free of contaminating leukocytes, and because of its operational simplicity, large numbers of blood samples can be processed. Our procedure also compares well with those using expensive commercial Sepacell resins in its ability to separate leukocytes from whole blood. As a test of usefulness in molecular biologic investigations, the parasites obtained from the blood of malaria-infected patients using the modified procedure yield genomic DNA whose single copy gene, the circumsporozite gene, efficiently amplifies by polymerase chain reaction.

Esparganose em alguns vertebrados do Brasil: dificuldades na identificaçao das espécies de Luheella (Spirometra)

Some species of Amphibia and Reptilia are listed as new hosts of spargana, from material deposited in the Helminthological Collection of Oswaldo Cruz Institute. It is discussed the difficulties in identifying the larvae (Sparganum) and also the identification of adults of Luheella species from South America. The histopathology induced by spargana in the liver of a species of Amphibia is briefly described.

Natural infection of wild rodents by Schistosoma mansoni parasitological aspects

The evaluation of the role of rodents as natural hosts of Schistosoma mansoni was studied at the Pamparrão Valley, Sumidouro, RJ, with monthly captures and examination of the animals. Twenty-three Nectomys squamipes and 9 Akodom arviculoides with a shistosomal infection rate of 56.5% and 22.2% respectively eliminated a great majority of viable eggs. With a strain isolated from one of the naturally infected N. squamipes, we infected 75% of simpatric Biomphalaria glabrata and 100% of albino Mus musculus mice. The adult worms, isolated from N. squamipes after perfusion were located mainly in the liver (91.5%) and the mesenteric veins (8.5%). The male/female proportion was 2:1. The eggs were distributed on small intestine segments (proximal, medial and distal portions) and the large intestine without any significant differences in egg concentration of these segments. In A. arviculoides, the few eggs eliminated by the stools were viable and there was litlle egg retention on intestinal segments. Considering the ease to complete S. mansoni biological cycle in the Nectomys/Biomphalaria/Nectomys system under laboratory conditions, probably the same is likely to occur in natural conditions. In support to this hypotesis there are also the facts that human mansonic shistosomiasis has a very low prevalence in Sumidouro and endemicity among the rodents has not changed even after repetead treatments of the local patients. Based on our experiments, we conclude that N. squamipes has become a natural host of S. mansoni and possibly may participate in keeping the cycle of schistosomiasis transmission at Pamparrão Valley.

Simian malaria in Brazil

In Brazil simian malaria is widely spread, being frequent in the Amazon region (10% of primates infected) and even more in the forested coastal mountains of the Southeastern and Southern regions (35% and 18% infected, respectively), but absent in the semi-arid Northeast. Only two species of plasmoidia have been found: the quartan-like Plasmodium brasilianum and the tertian-like P. simium, but the possible presence of other species is not excluded. P. brasilianum is found in all enzootic foci, but P. simium was detected only on the coast of the Southeastern and Southern regions, between parallels 20-S and 30-S. Nearly all hosts are monkeys (family Cebidae, 28 species harbouring plasmodia out of 46 examined) and very rarely marmosets or tamarins (family Callitrichidae, I especies out of 16). P. brasilianum was present in all infected species, P. simium in only two. The natural vector in the Southeastern and Southern regions was found to be Anopheles cruzi, but has not been conclusively identified in the Amazon. One natural, accidental human infection due to P. simium was observed. There is no evidence of the relation of the simian to human malaria in the Southeastern and Southern regions, where human malaria was eradicated in spite of the high rates of monkeys infected, but in the Amazon recent serological studies by other workers, revealing high positivity for P. brasilianum/P. malariae antibodies in local indians, would suggest that among them malaria might be regarded as a zoonosis.

A study on the pathogenesis of human cerebral malaria and cerebral babesiosis

Cerebral complications are important, but poorly understood pathological features of infections caused by some species of Plasmodium and Babesia. Patients dying from P. falciparum were classified as cerebral or non-cerebral cases according to the cerebral malaria coma scale. Light microscopy revealed that cerebral microvessels of cerebral malaria patients were field with a mixture of parazited and unparazited erythrocytes, with 94% of the vessels showing parasitized red blood cell (PRBC) sequestration. Some degree of PRBC sequestration was also found in non-cerebral malaria patients, but the percentage of microvessls with sequestered PRBC was only 13% Electron microscopy demonstrated knobs on the membrane of PRBC that formed focal junctions with the capillary endothelium. A number of host cell molecules such as CD36, thrombospondim (TSP) and intracellular adhesion molecule I (ICAM-1) may function as endothelial cell surfacereports for P. falciparum-infected erythrocytes. Affinity labeling of CD36 and TSP to the PRBC surface showed these molecules specifically bind to the knobs. Babesia bovis infected erythrocytes procedure projections of the erythrocyte membrane that are similar to knobs. When brain tissue from B. bovis-infected cattle was examined, cerebral capillaries were packed with PRBC. Infected erythrocytes formed focal attachments with cerebral endothelial cells at the site of these knob-like projections. These findings indicate that cerebral pathology caused by B. bovis is similar to human cerebral malaria. A search for cytoadherence proteins in the endothelial cells may lead to a better understanding of the pathogenisis of cerebral babesiosis.

Protection of aouts monkeys after immunization with recombinant antigens of Plasmodium falciparum

The genus Aotus spp. (owl monkey) is one of the WHO recommended experimental models for Plasmodium falciparum blood stage infection, especially relevant for vaccination studies with asexual blood stage antigens of this parasite. For several immunization trials with purified recombinant merozoite/schizont antigens, the susceptible Aouts kenotypes II, III, IV and VI were immunized with Escherichia coli derived fusion proteins containg partial sequences of the proteins MSAI (merozoite surface antigen I), SERP (serine-strech protein) and HRPII (histidine alanine rich protein II) as well as with a group of recombinant antigens obtained by an antiserum raised against a protective 41 kD protein band. The subcutaneous application (3x) of the antigen preparations was carried out in intact animals followed by splenectomy prior to challange, in order to increase the susceptibility of the experimental hosts to the parasite. A partial sequence of HRPII, the combination of three different fusion proteins of the 41 kD group and mixture of two sequences of SERP in the presence of the modified Al(OH)3 adjuvant conferred significant protection against a challange infection with P. falciparum blood stages (2-5 x 10 (elevado a sexta potência) i. RBC). Monkey immunized with the MS2-fusion protein carrying the N-terminal part of the 195 kD precursor of the major merozoite surface antigens induced only marginal protection showing some correlation between antibody titer and degree of parasitaemia. Based on the protective capacity of these recombinant antigens we have expressed two hybrid proteins (MS2/SERP/HRPII and SERP/MSAI/HRPII) in E. coli containing selected partial sequences of SERP, HRPII and MSAI. Antibodies raised against both hybrid proteins in rabbits and Aotus monkeys recognize the corresponding schizont polypeptides. In two independent immunization trials using 13 animals (age 7 months to 3 years) we could show that immunization of Aotus monkeys with either of the two hybrid proteins administered in an oil-based well tolerated formulation protected the animals frm a severe experimental P. falciparum (strain Palo Alto) infection.

The susceptibility of adult schistosomes to immune attrition

Mouse infection models are described that demonstrate reduction of egg production in Schistosoma haematobium infections and both worm loss and reduced fecundity in S. bovis infections. Neither phenomenum could be shown in S. mansoni infected mice. The immunological basis for these anti-adult responses was inferred by comparison with infections in T-cell deprived mice and by the serum transfer of the ability to reduce a S. bovis worm burden into immunocompromised hosts. Vaccination with irradiation attenuated parasites was also shown to have consequences for the adults of a challenge infections of S. haematobium and S. bovis specifically. Prior vaccination resulted in an abrogation of the anti-fecundity and adult worm elimination that occurred in non-vaccinated similary infected mice. hese models are being used to define the targets and mechanisms involved in anti-adult attrition. A serological assay, quantitation of a circulating antigen (CAA) has been assessed for its ability to measure worm burdens of different species of schistosome in mice. This assay will be used to question whether anti-adult immunity contributes to the pattern of infection with S. mansoni and S. haematobium in man.

Experimental murine schistosomiasis mansoni: estabilishment of the chronic phase of the disease

After the acute hyperergic phase of schistosomal infection, the chronic phase of the disease corresponds to the estabilishment of a relative equilibrium between the host and the parasite. This involves: (1) A shift from the predominance of the TH2 response observed in the acute phase, to the predominance of the TH1 response in the chronic phase of the disease, with modification of lymphokine and immunoglobulin secretions patterns. (2) Redistribution of hosts responses to parasite, with predominance of systemic controls in the acute phase, and a shift towards local tissue responses in the chronic phase. This redistribution relieves the hyperergic responses involving the whole body of the host, and delimits cellular and molecular reactions to parasites to only those tissues that are directly involved by the adult parasites and their eggs. Mobilization of eosinophil granulocytes in schistosomal periovular granulomas is one of examples of this redistribution.

Ecological studies on the intermediate host snails and the relevance to schistosomiasis control

A detailed knoledge of distribution patterns schistosome intermediate hsts and their populations dynamics and factors affecting these patterns will provide useful information about the possibilities and desirability of conducting snail control measures in various transmission situations. On the basis of various case studies the association between the occurence of human water contacts and the presence of schistosome intermediate hosts or infections in the intermediate hosts is illustrated. Other parameters affecting snail distribution patterns and density fluctuations are discussed. It is concluded that ecological studies on the intermediate host are extremely relevant, either to optimally apply existing control measures or to develop alternative measures of snail control, such as ecological or biological control.

The distribution of intestinal helminth infections in a rural village in Guatemala

Fecal egg count scores were used to investigate the distribution and abundance of intestinal helminths in the population of a rural village. Prevalences of the major helminths were 41% with Ascaris lumbricoides 60% with Trichuris trichiura and 50% with Necator americanus. All three parasites showed a highly aggregated distribution among hosts. Age/prevalence and age/intensity profiles were typical for both A. lumbricoides and T. trichiura with the highest worm burdens in the 50-10 year old children. For hookworm both prevalence and intensity curves were convex in shape with maximum infection levels in the 30-40 year old age class. Infected females had higher burdens of T. trichiura than infected males in all age classes of the population; there were no other effects of host gender. Analysis of associations between parasites within hosts revealed strong correlations between A. lumbricoides and T. lumbricoides and T. trichiura. Individuals with heavy infections of A. lumbricoides and T. trichiura showed highly significant aggregation within households. Associations between a variety of household features and heavy infections with A. lumbricoides and T. trichiura are described.

Adult abundance, biting behavior and parity of Anopheles aquasalis, Curry 1932 in two malarious areas of Sucre State, Venezuela

The principal vector of malaria in eastern Venezuela, Anopheles aquasalis, is exophagic and exophilic. Control using indoor insecticide house sprays has failed to lower the number of malaria cases. Therefore, studies were initiated in two villages of the eastern coastal state of Sucre to better understand this vector's biology and develop a more integrated control program. An. aquasalis was found to have a crepuscular biting behavior with a major peak at dusk and a minor peak at dawn. Mosquitos were collected more outdoors than indoors. Forty-seven percent of the biting took place before people went to bed (22:30 hr) and 69% of the mosquitos biting during this time period bite outdoors. Outdoor biting could be the reason why indoor spraying alone did not lower malaria cases. Seasonal abundance was greater in the rainy season compared to the dry season. Seasonal parous rates were high (78.3%-100%) and similar indoors and outdoors and between dry and wet season in Santa F e. In Guayana, the seasonal parity was lower (34.6%-42.2%) than Santa F e with indoor parity slightly higher than outdoors. Malaria cases were higher in Santa F e, but adult mosquito density was much lower than in Guayana. This difference could have been due to higher parity in Santa F e compared to Guayana. The greater distance to the nearest breeding site and presence of alternative hosts in Guayana can not be discounted as factors which contributed to the difference in malaria transmission between locations. We concluded that knowledge on seasonal occurrence, biting activity, resting behavior and breeding site location can be used to design a new control strategy for this vector.

Survey of the irp2 gene among Yersinia pestis strains isolated during several plague outbreaks in northeast Brazil

The irp2 gene codes for a 190 kDa protein (HMWP2) synthesidez when highly pathogenic Yersinia are grown under conditions of iron starvation. In this work, the presence of irp2 in strains of Y. pestis isolated from different hosts during several plague outbreaks in the foci of Northeast Brazil wasstudied. For this purpose, 53 strains were spotted onto nylon filters and their DNA was hybridized with the A13 probe which is a 1 kb fragment of the irp2 coding sequence. All strains except two hybridized with the probe. However, when the initial stock culture of these two strains were analyzed, they both proved to bepositive with the A13 probe, indicating that the locus was lost after subculturein vitro but was always present in vivo. To examine the degree of conservation of the chromosomal fragment carrying irp2 among Brazilian strains, the hybridization profiles of 15 strains from different outbreaks, different hosts and different foci were compared. The hybridization profiles of these strains were all identical when their DNA was digested with either EcoRI, EcorRV or AvaII, indicatingthat the restriction sites surrounding the irp2 locus are very well conserved among Northeast Brazilian strains of Y. pestis. Altogether, these results suggest that the irp2 chromosomal region should be of prime importance for the bacteria during their multiplication in the host.

IL-13 induces expression of CD36 in human monocytes through PPARgamma activation.

The class B scavenger receptor CD36 is a component of the pattern recognition receptors on monocytes that recognizes a variety of molecules. CD36 expression in monocytes depends on exposure to soluble mediators. We demonstrate here that CD36 expression is induced in human monocytes following exposure to IL-13, a Th2 cytokine, via the peroxisome proliferator-activated receptor (PPAR)gamma pathway. Induction of CD36 protein was paralleled by an increase in CD36 mRNA. The PPARgamma pathway was demonstrated using transfection of a PPARgamma expression plasmid into the murine macrophage cell line RAW264.7, expressing very low levels of PPARgamma, and in peritoneal macrophages from PPARgamma-conditional null mice. We also show that CD36 induction by IL-13 via PPARgamma is dependent on phospholipase A2 activation and that IL-13 induces the production of endogenous 15-deoxy-Delta12,14-prostaglandin J2, an endogenous PPARgamma ligand, and its nuclear localization in human monocytes. Finally, we demonstrate that CD36 and PPARgamma are involved in IL-13-mediated phagocytosis of Plasmodium falciparum-parasitized erythrocytes. These results reveal a novel role for PPARgamma in the alternative activation of monocytes by IL-13, suggesting that endogenous PPARgamma ligands, produced by phospholipase A2 activation, could contribute to the biochemical and cellular functions of CD36.