Study of some parameters affecting the in vitro cultivation of Plasmodium falciparum within saimiri sciureus red blood cells

The in vitro growth and multiplication of the erythrocytic stages of Plasmodium falciparum within Saimiri sciureus (squirrel monkey) red blood cells have been studied. Various parameters, such as the origin of the red blood cells and serum supplement, nature of the buffer, influence of the final pH of the medium, role of proteose peptone and glucose addition, were investigated. The selection of the best culture conditions led to the obtention of a reproducible in vitro growth of two parasite cycles in Saimiri erythrocytes, which is an useful achievement for in vitro studies. Our failure to establish a continuous culture line for longer than 19 days, could be explained by a dramatic increasing of osmotic fragility of the Saimiri red blood cells related to their small size.

A review of achievements of the national schistosomiasis control program in middle and upper Egypt areas

In Egypt the "national schistosomiasis control program" was formulated to control transmission by reduction of prevalence and intensity of current infections, and thereby achieve an acceptable level of schistosomiasis disease control. The program was implemented foremost in Middle Egypt (1977) and Upper Egypt (1980), collectively extending 800 km alongside of the River Nile and accommodate about 10.5 million people. Schistosoma haematobium has been essentially the prevailing species infection in both areas. The strategy of control entailed both area-wide mollusciciding with niclosamide, and selective population chemotherapy with metrifonate. Evaluation in 1986 showed that prevalence dropped from pre-control 29.4% in Middle Egypt and 26.3% in Upper Egypt to 6% and 7.8% respectively, together with a remarkable drop of infections among children. Also mean intensity attained low levels consistent of low grade infections. It is evident therefore that in these areas where an enhancement of schistosomiasis infections had been anticipated the employment of the twofold strategy effected a state of low-prevalence/low-intensity signifying a lowered reservoir of infection and a substantial interference with the potentials of transmission.

The brazilian leishmaniasis control program

The control methods used in Brazil for both cutaneous and visceral leishmaniasis by the Ministry of Health's National Health Foundation (Fundação Nacional de Saúde) are presented. Data referring to the programme between 1980 and 1991 is presented. There was an increase in the total number of notified cases of cutaneous leishmaniasis until 1987 and from this date onwards the number has remained between 23,000 and 26,000 per year. There were significantly fewer cases of visceral leishmaniasis during the same period and until 1991 the greatest number of cases reported in one year was 2,511 in 1985. The overall picture is that it is difficult to evaluate to what extent the control methods employed have reduced the number of cases of either form of the disease. It is suggested that there is an urgent need for a closer approximation of research and health workers to review the actual control strategies and to define procedures capable of assessing accurately their impact.

Biological control program against simuliids in the State of São Paulo, Brazil

In Brazil, the use of biological vector-control methods has been largely confined to experimental research, with little or no application of such techniques by public institutions responsible for implementing control programs. The notable exceptions have been the black fly control program carried out by the Health Secretariat in the State of São Paulo. Since the 1980s, São Paulo's "Superintendência de Controle de Endemias" has been conducting studies on the viability of using Bacillus thuringiensis (H-14) for simuliid control, and the results have been so encouraging that the agency has now incorporated this method into its Simuliid Control Program.

Thermal effect on the life-cycle parameters of the medically important freshwater snail species Lymnaea (Radix) luteola (Lamarck)

The snails Lymnaea (Radix) luteola exhibited marked variations in growth, longevity, and attaining sexual maturity at different temperatures and diets. At 10°C, irrespective of foods, pH and salinity of water, the snails had minimum life span, maximum death rate and lowest growth rate. At 15°C, the growth rate was comparatively higher and the snails survived for a few more days. But at these temperatures they failed to attain sexual maturity. Snails exposed to pH 5 and 9 at 20°, 25°, 30°, 35°C and room temperatures (19.6°-29.6°C); to 0.5, 1.5 and 2.5 NaCl ‰ at 20° and 35ºC; to 2.5 NaCl ‰ at 25°C and room temperatures failed to attain sexual maturity. The snails exposed to pH 7 and different salinity grades at 20°, 25°, 30°, 35°C and room temperatures became sexually mature between 25-93 days depending upon the type of foods used in the culture.

Update of the Gene Discovery Program in Schistosoma mansoni with the Expressed Sequence Tag Approach

Continuing the Schistosoma mansoni Genome Project 363 new templates were sequenced generating 205 more ESTs corresponding to 91 genes. Seventy four of these genes (81%) had not previously been described in S. mansoni. Among the newly discovered genes there are several of significant biological interest such as synaptophysin, NIFs-like and rho-GDP dissociation inhibitor

Biological Parameters and Molecular Markers of Clone CL Brener - The Reference Organism of the Trypanosoma cruzi Genome Project

Clone CL Brener is the reference organism used in the Trypanosoma cruzi Genome Project. Some biological parameters of CL Brener were determined: (a) the doubling time of epimastigote forms cultured in liver infusion-tryptose (LIT) medium at 28oC is 58±13 hr; (b) differentiation of epimastigotes to metacyclic trypomastigotes is obtained by incubation in LIT-20% Grace´s medium; (c) trypomastigotes infect mammalian cultured cells and perform the complete intracellular cycle at 33 and 37oC; (d) blood forms are highly infective to mice; (e) blood forms are susceptible to nifurtimox and benznidazole. The molecular typing of CL Brener has been determined: (a) isoenzymatic profiles are characteristic of zymodeme ZB; (b) PCR amplification of a 24Sa ribosomal RNA sequence indicates it belongs to T. cruzi lineage 1; (c) schizodeme, randomly amplified polymorphic DNA (RAPD) and DNA fingerprinting analyses were performed

Comparison of some behavioral and physiological feeding parameters of Triatoma infestans klug, 1834 and Mepraia spinolai porter, 1934, vectors of chagas disease in Chile

There are two vectors of Chagas disease in Chile: Triatoma infestans and Mepraia spinolai. We studied the feeding behavior of these species, looking for differences which could possibly explain the low impact of the latter species on Chagas disease. Both species used thermal cues to locate their feeding source and consumed a similar volume of blood which was inversely related to the body weight before the meal and directly related to the time between meals. The average time between bites were 6.24 and 10.74 days. The average bite of M. spinolai lasted 9.68 min, significantly shorter than the 19.46 min for T. infestans. Furthermore, while T. infestans always defecated on the host, this behavior was observed in M. spinolai in only one case of 27 (3.7%). The delay between the bites and defecation was very long in M. spinolai and short in T. infestans. These differences may affect the reduced efficiency of transmission of Chagas infection by M. spinolai.

Simulium spp. control program in Rio Grande do Sul, Brazil

Insects of the Simuliidae family have been the object of control in Rio Grande do Sul since the 70s. Their constant attacks became a social-economical problem as well as a problem of Public Health, with serious consequences to men and to the economy of the areas in which the insects develop. At first, the control was done with a chemical larvicide Themephos ABATE 500 E, but an imperfect measuring of outflow to determine the quantity of the product made Simulium spp. resistant to it. From 1983 on, following a study of a new method for the outflow measuring, we started to use a biological larvicide Bacillus thuringiensis serovar israelensis based. The biological control uses the new method in 36.4% of the state area, assisting about 3,500,000 inhabitants.

Life cycle and reproductive parameters of Clerada apicicornis signoret (Hemiptera: Lygaeidae) under laboratory conditions

The life cycle of Clerada apicicornis was determined under laboratory conditions. Mean development times in days were: egg 27.2, nymph I 12.5, nymph II 12, nymph III 13.4, nymph IV 16.4, nymph V 26. The life expectancy of adults ranged from 117 to 317 days (mean 196 days). Based on a cohort of 29 females of C. apicicornis, a horizontal life table was constructed. The following predictive parameters were obtained: net rate of reproduction (Ro = 48.31), intrinsic rate of population increase (r m = 0.153), generation time (Tc = 28.20 weeks), and finite rate of population increment (lambda = 1.16). The reproductive value (Vx) for each age class of the cohort females was calculated. The following observed parameters were calculated after mortality in each stage: net rate of reproduction (R'o=13.4), intrinsic rate of population increase (r c' =0.09 ), and finite rate of population increment (lambda' =1.1). The generation time (Tc' =27.4) was estimated using the methods of Laughlin and Bengstron. A vertical life table was elaborated and mortality was described for one generation of the cohort.

Schistosomiasis mansoni: follow-up of control program based on parasitologic and serologic methods in a Brazilian community of low endemicity

A field survey on schistosomiais was carried out in 1998, in the municipality of Pedro de Toledo, a low endemic area in the state of São Paulo, Brazil. According to the parasitologic Kato-Katz method, the prevalence rate was 1.6%, with an infection intensity of 40.9 eggs per gram of stool. By the immunofluorescence test (IFT) for detection of IgG and IgM antibodies in the serum, IgG-IFT and IgM-IFT, respectively, prevalence indices of 33.2% and 33.5% were observed. To assess the impact of the schistosomiasis control program in the area, parasitologic and serologic data obtained in 1998, analyzed according to the age, sex, and residence zone, were compared to previous data obtained in a epidemiologic study carried out in 1980, when prevalence indices were of 22.8% and 55.5%, respectively by Kato-Katz and IgG-IFT. A significant fall of the prevalence was observed, indicating that the control measures were effective. Nonetheless, residual transmission was observed, demonstrating the need for a joint effort to include new approaches for better understanding the real situation and improving the control of the disease in low endemic areas.

Parameters affecting cellular invasion and escape from the parasitophorous vacuole by different infective forms of Trypanosoma cruzi

In this study we have examined certain aspects of the process of cell invasion and parasitophorous vacuole escape by metacyclic trypomastigotes and extracellular amastigote forms of Trypanosoma cruzi (G strain). Using Vero (and HeLa) cells as targets, we detected differences in the kinetics of vacuole escape by the two forms. Alcalinization of intercellular pH influenced both invasion as well as the escape from the parasitophorous vacuole by metacyclic trypomastigotes, but not the escape kinetics of extracellular amastigotes. We used sialic acid mutants as target cells and observed that the deficiency of this molecule facilitated the escape of both infective forms. Hemolysin activity was only detected in extracellular amastigotes and neither form presented detectable transialidase activity. Invasion of extracellular amastigotes and trypomastigotes in Vero cells was affected in different ways by drugs that interfere with host cell Ca2+ mobilization. These results are in line with previous results that indicate that metacyclic trypomastigotes and extracellular amastigote forms utilize mechanisms with particular features to invade host cells and to escape from their parasitophorous vacuoles.

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.

Antimicrobial susceptibility patterns of unusual nonfermentative gram-negative bacilli isolated from Latin America: report from the SENTRY Antimicrobial Surveillance Program (1997-2002)

The antimicrobial susceptibility of 176 unusual non-fermentative gram-negative bacilli (NF-GNB) collected from Latin America region through the SENTRY Program between 1997 and 2002 was evaluated by broth microdilution according to the National Committee for Clinical Laboratory Standards (NCCLS) recommendations. Nearly 74% of the NF-BGN belonged to the following genera/species: Burkholderia spp. (83), Achromobacter spp. (25), Ralstonia pickettii (16), Alcaligenes spp. (12), and Cryseobacterium spp. (12). Generally, trimethoprim/sulfamethoxazole (MIC50, < 0.5 µg/ml) was the most potent drug followed by levofloxacin (MIC50, 0.5 µg/ml), and gatifloxacin (MIC50, 1 µg/ml). The highest susceptibility rates were observed for levofloxacin (78.3%), gatifloxacin (75.6%), and meropenem (72.6%). Ceftazidime (MIC50, 4 µg/ml; 83.1% susceptible) was the most active beta-lactam against B. cepacia. Against Achromobacter spp. isolates, meropenem (MIC50, 0.25 µg/ml; 88% susceptible) was more active than imipenem (MIC50, 2 µg/ml). Cefepime (MIC50, 2 µg/ml; 81.3% susceptible), and imipenem (MIC50, 2 µg/ml; 81.3% susceptible) were more active than ceftazidime (MIC50, >16 µg/ml; 18.8% susceptible) and meropenem (MIC50, 8 µg/ml; 50% susceptible) against Ralstonia pickettii. Since selection of the most appropriate antimicrobial agents for testing and reporting has not been established by the NCCLS for many of NF-GNB species, results from large multicenter studies may help to guide the best empiric therapy.

Experimental infection parameters in Galea spixii (Rodentia: Caviidae) with Leishmania infantum chagasi

In order to better understand the epidemiological transmission network of leishmaniasis, an endemic disease in Northeast Brazil, we investigated the susceptibility of Spix yellow-toothed cavies (Galea spixii) to the Leishmania infantum chagasi parasite. Nine cavies were experimentally infected, separated into three groups and monitored at 30, 90 and 180 days, respectively. Amastigotes were identified in the spleen slides of two cavies killed 180 days after infection. Antibodies against the L. i. chagasi were identified in one of the cavies. This demonstrates that G. spixii is in fact capable of maintaining a stable infection by L. i. chagasi without alterations in biochemical and hematological parameters of the host and without perceivable micro and macroscopic lesions.