SJ Strain Research Articles

Isolation and identification of a novel ginger tissue pathogenic bacteria Citrobacter sp. SJ7

The pathogenic bacteria of ginger plantlets were isolated and identified. Pathogenic strains were isolated, purified, and then identify by morphology observation, physiological and biochemical experiments, and the 16s rDNA sequence was analyzed to verify the classification of the pahtogen. This study shows the novel pathogenic bacteria is kind of Gram-negative ones, which can be well root on the ginger tissue as well as cultivated plantlets. The histopathology examination illustrates that the pathogen prefer infect the host plant through ginger tuber. Moreover, the16S rDNA sequence analysis exhibits that it is kind homology species closely relating to Citrobacter amalonaticus. In a word, we first isolated a novel pathogenic bacteria of ginger named as Citrobacter sp SJ strain, which could successfully recognize the host ginger via xylems.

Two strains of roof rats as effective models for assessing new-object reaction.

Wild animals generally avoid even small and harmless novel objects and/or familiar objects moved to a novel position, which is termed “new-object reaction”. Although new-object reaction appears to be a biologically important characteristic for animals, little progress has been made in understanding the neural mechanisms underlying new-object reaction. One reason might be the lack of effective experimental animals. Two strains of roof rats (Sj and Og strains) were established from wild roof rats caught in Shinjuku, Tokyo and one of the Ogasawara Islands, respectively, by a Japanese pest control company. Based on the rat caregivers’ informal observations, we conducted behavioral and anatomical tests to assess the validity of Sj and Og strains for the analyses of new-object reaction. In Experiment 1, the Sj strain showed reduced food consumption compared with the Og strain when food was provided in a novel way, suggesting that the Sj strain had a stronger avoidance of novel objects compared with the Og strain. Experiment 2 demonstrated that the basolateral complex of the amygdala and bed nucleus of the stria terminalis in experimental Sj rats had a larger percentage area compared with that of experimental Og rats, indicating these nuclei might be involved in the difference observed in avoidance of novel objects between the strains. Taken together, the present study suggests that Sj and Og strains are effective experimental animals for assessing new-object reaction.

Host-parasite relationship between Biomphalaria amazonica (Paraense, 1966) and Schistosoma mansoni (Sambon, 1907).

Biomphalaria amazonica is a planorbid species considered a potential host of Schistosoma mansoni. It is widely distributed in the Neotropical zone, particularly in the North and Centre-West of Brazil and in the North of Bolivia. The aim of the present study was to determine the host-parasite relationship between B. amazonica and S. mansoni (BH and SJ strains). Specimens of B. amazonica and their snail-conditioned water were examined in terms of their ability to attract miracidia. The infectivity of the mollusks was determined by exposing them to 20 miracidia of both strains. Sporocyst development and amebocyte reactions were studied after each mollusk specimen was exposed to 100 miracidia. Although no cercariae were eliminated, specimens of B. amazonica proved capable of attracting 77% of the miracidia they were exposed to. Viable sporocysts with no amebocyte reaction were found 96 hours after the exposure to miracidia. These results indicate the susceptibility of B. amazonica to the BH and SJ strains of S. mansoni, and therefore demonstrate the importance of this planorbid species as a potential vector of the trematode in the areas where it occurs.

A Study of the Granulomatous Responses Induced by Different Strains ofSchistosoma mansoni

The increased pathogenesis of the Schistosoma mansoni BH strain compared with the SJ strain has been attributed to the number of granulomas formed in experimental infections, which increase the mortality in definitive hosts. The aim of the present study was to investigate the development of granulomas around the eggs of the S. mansoni BH and SJ strains and to determine whether this host reaction was strain specific. Four experimental groups were analyzed. Two groups contained mice inoculated in the caudal vein with eggs from the S. mansoni BH or SJ strains and the other two contained mice that were infected with cercariae of the BH strain prior to being inoculated with eggs. The number of granulomas per tissue area in the lungs and liver, as well as the size of the granulomas, was analyzed to characterize the response to schistosome infection. The largest granulomatous responses were observed around eggs of the BH strain. Granulomas covered a larger area in the lungs of mice that were previously infected with cercariae and subsequently inoculated with eggs of the BH strain. These results indicated that specific granulomatous responses occurred following an infection with the BH and SJ strains of S. mansoni.

Experimental Evaluation of the Pathogenicity of Different Strains ofSchistosoma mansoni

The pathogenesis of three different Schistosoma mansoni strains from the Brazilian states of Minas Gerais (BH strain) and São Paulo (SJ and SD strains) was evaluated in experimentally infected mice. Observations of the most severe clinical cases among local patients treated (SD strain) in the city of Campinas (São Paulo, Brazil) formed the basis of this study. Mice were used as definitive hosts and were infected with cercariae from Biomphalaria tenagophila (SJ and SD strains) and Biomphalaria glabrata (BH strains). The parameters analyzed were as follows: number of S. mansoni eggs in mice feces; number of granulomas per tissue area in liver, spleen, lungs, pancreas, and ascending colon; measurements of hepatic and intestinal granulomas; number of adult worms; and measurements of trematode eggs. The comparison among the three strains indicated that the SD strain, isolated in Campinas, presented a higher worm recovery relative to the number of penetrating cercariae. In addition, when compared to the SJ and BH strains, the SD strain demonstrated similar pathogenicity to the BH strain, with a greater quantity of granulomas in the viscera, as well as larger granulomas and eggs. Furthermore, a greater quantity of trematode eggs was also shed in the feces.

Hybridism between Biomphalaria cousini and Biomphalaria amazonica and its susceptibility to Schistosoma mansoni

Molecular techniques can aid in the classification of Biomphalaria species because morphological differentiation between these species is difficult. Previous studies using phylogeny, morphological and molecular taxonomy showed that some populations studied were Biomphalaria cousini instead of Biomphalaria amazonica. Three different molecular profiles were observed that enabled the separation of B. amazonica from B. cousini. The third profile showed an association between the two and suggested the possibility of hybrids between them. Therefore, the aim of this work was to investigate the hybridism between B. cousini and B. amazonica and to verify if the hybrids are susceptible to Schistosoma mansoni. Crosses using the albinism factor as a genetic marker were performed, with pigmented B. cousini and albino B. amazonica snails identified by polymerase chain reaction-restriction fragment length polymorphism. This procedure was conducted using B. cousini and B. amazonica of the type locality accordingly to Paraense, 1966. In addition, susceptibility studies were performed using snails obtained from the crosses (hybrids) and three S. mansoni strains (LE, SJ, AL). The crosses between B. amazonica and B. cousini confirmed the occurrence of hybrids. Moreover, hybrids can be considered potential hosts of S. mansoni because they are susceptible to LE, SJ and AL strains (4.4%, 5.6% and 2.2%, respectively). These results indicate that there is a risk of introducing schistosomiasis mansoni into new areas.

Influence of saccharose on the development of cercariae from Schistosoma mansoni strains BH and SJ

The development of cercariae from Schistosoma mansoni strains BH and SJ in Biomphalaria glabrata and Biomphalaria tenagophila treated with saccharose was studied. The molluscs were maintained in dechlorinated tap water containing 0.01% saccharose. After one week of treatment with saccharose, B. glabrata and B. tenagophila were exposed to ten S. mansoni miracidia, from BH and SJ strains respectively. Control snails of both species were maintained in dechlorinated tap water without saccharose and exposed to the same number of miracidia. There was no significant difference between the infection rates of snails treated or not with saccharose. However, the two groups of B. glabrata had significantly greater infection rates than the corresponding B. tenagophila groups. Molluscs treated with saccharose had a lower survival rate, with the greatest mortality occurring immediately before and at the beginning of cercariae release. Treatment with saccharose did not result in the release of more cercariae, but larvae from molluscs so treated showed a greater capacity to penetrate mouse skin, which was attributed to the greater energy supply during larval development in the mollusc.

Biomphalaria tenagophila (Orbigny, 1835) (Mollusca): adaptation to desiccation and susceptibility to infection with Schistosoma mansoni Sambon, 1907.

Experiments were carried out to test the susceptibility of Biomphalaria tenagophila to the infection with strain SJ of Schistosoma mansoni in the F1, F2 and non-selected parental generation. The potential adaptation of B. tenagophila to desiccation, in healthy mollusks and those exposed to the larvae of S. mansoni of the F1, F2 and non-selected parental generations was also studied. The presence of mucus and soil, at the shell opening, protected the snails against desiccation, favoring survival. The healthy mollusks performed more attempts against desiccation than those exposed to the larvae of the parasite. The mortality rate, during desiccation, was higher among mollusks that remained buried and with the shell opening unobstructed. During the desiccation period the stage of development of the parasite was influenced by the weight loss and the survival of the snails. The longer the period of desiccation, the greater was the weight loss observed, abbreviating survival. The non-selected parental generation was more sensitive to desiccation than the F1 and F2 generations, both in healthy mollusks and in those exposed to S. mansoni larvae. Healthy mollusks were more resistant to desiccation than those exposed to the larvae of the S. mansoni. Desiccation did not interrupt the development of S. mansoni larvae in mollusks, causing a delay in the cercariae elimination. The susceptibility of B. tenagophila to the SJ strain of S. mansoni, in mollusks maintained in water during the larvae incubation period, was similar in all three generations.

Resistance of Biomphalaria occidentalis from Varzea das Flores dam, Minas Gerais, to Schistosoma mansoni infection detected by low stringency polymerase chain reaction.

Biomphalaria occidentalis Paraense, 1981 from Varzea das Flores dam, MG, Brazil, was exposed to infection with Schistosoma mansoni. Individual infection was performed with 140 B. occidentalis and 100 B. glabrata snails using LE and SJ strains. Two groups of B. occidentalis were killed after seven day-miracidia exposure to detect S. mansoni DNA, through the low stringency polymerase chain reaction (LS-PCR), and were negative. The infection rates were 69.2% (LE strain) and 96.7% (SJ strain) for B. glabrata and 0% for B. occidentalis. LS-PCR enabled early resistance diagnosis.

Differences in Brazilian strains of Schistosoma mansoni evaluated by means of morphometric analysis of cercariae of both sexes.

Morphometrics of Brazilian strains (BH, SJ and CMO) of Schistosoma mansoni cercariae were obtained with a computerized image analyzer (IMAGE PRO PLUS, MEDIA CYBERNETICS), considering the following characters: body area, tail, furcae, oral and ventral suckers and distance between them. For statistical analysis, the variance test (one-way Anova) was applied and significant differences of p< 0.05 were considered. All morphometric values in the BH strain were significantly higher (p< 0.05) than in the others. Lower values were obtained in females of SJ strain for all characters, excepting the body area. Only this character showed to be significantly different in males and females of the three strains. Specimens of both sexes in the BH and SJ strains showed significant differences regarding all characters. It was observed that this morphometric analysis permits the characterization of strains and also the sex identification in S. mansoni cercariae. Due to its feasibility, this method can be applied as a tool in laboratories devoid of more complex equipment.

Schistosoma mansoni: Continuous Variation in Susceptibility of the Vector Snail of Schistosomiasis, Biomphalaria tenagophila I. Self-Fertilization—Lineage

Mascara, D., Kawano, T., Magnanelli, A. C., Silva, R. P. S., Sant' Anna, O. A., and Morgante, J. S. 1999. Schistosoma mansoni: continuous variation in susceptibility of the vector snail of schistosomiasis, Biomphalaria tenagophila I. Self-Fertilization—Lineage. Experimental Parasitology93, 133–141. Artificial selection of Biomphalaria tenagophila snails for susceptibility to infection by Schistosoma mansoni (Brazilian SJ strain) was carried out from natural populations. After five self-fertilization generations, two lineages were isolated and were designated as SUSC (highly susceptible 93–100%) and RES (nonsusceptible 5–0%). Length of the prepatent period, cercarial production, and mortality of the hosts in postexposure were determined in all generations (F1–F8) and were analyzed as quantitative traits related to host susceptibility. Distribution patterns of frequencies were observed within snail families (samples derived from one F0 snail), these traits showing a significant influence by selection applied to susceptibility. The multiple quantitative classes were described in terms of continuous variation. During the selection of SUSC lineage, classes with higher values of prepatent length and lower cercarial production were eliminated, and the heritability calculated for these two traits was 0.811 and 0.709, respectively. Experimental results were correlated with an increase in the level of susceptibility in the generations selected and are discussed in relation to inheritance patterns as well as the quantitative variation of susceptibility.

Schistosoma mansoni: the sex ratios of worms in animals infected with cercariae from three species of Biomphalaria.

The ratios of male to female worms of Schistosoma mansoni were determined in mice infected with cercariae from LE, SJ and AL strains shed by mollusc hosts of the parasite in Brazil. The sex ratios of worms in the animals were similar with cercariae from Biomphalaria glabrata and B. tenagophila varying from 1.1:1 to 1.6:1 with LE and AL strains and 1:1.1 with SJ. In the animals infected with cercariae from B. straminea the ratio of male to female worms was similar to those obtained using cercariae shed from the other two species of molluscs, 1.5:1 with LE strain. Inoculations by AL and SJ cercariae resulted in sex ratios of 3.1:1 and 6:1 respectively. The normal sex ratios of worms established in Brazil in animals inoculated with cercariae from B. glabrata and B. tenagophila is from 1:1 to 2:1. The higher number of male worms that developed from cercariae of the AL and SJ strains obtained from B. straminea indicate a lower compatibility of the snail concerning these strains of S. mansoni.

Schistosoma mansoni: níveis de infecciosidade para os moluscos hospedeiros intermediários do Brasil

The levels of infectivity of Schistosoma mansoni for the three species of Biomphalaria, intermediate hosts of parasite in Brazil were studied after exposing of molluscs to miracidia in the laboratory and in the field. The LE and SJ strains of S. mansoni, maintained in laboratory were used in these experiments as well as the WVS and RFS strains obtained from faeces of schoolchildren from Belo Horizonte, MG. The results show the high level of infectivity of S. mansoni for B. glabrata with infection rates varying from 4.7 to 85.5%. The snail B. straminea was susceptible to LE, SJ and WVS strains, with infection rates of 11.0 to 24.6%, B. tenagophila was susceptible only to LE and SJ strains with infection rates of 2.5 to 6.5%. The mean number of cercariae of the WVS strain shed per day, by B. straminea and B. glabrata were 93 +/- 59 and 782 +/- 1,120, respectively.

Antischistosomal activity of acridanone-hydrazones in Cebus monkeys experimentally infected with the SJ strain of Schistosoma mansoni.

In this study, four compounds were utilized at the dose of 12.5 mg/kg body weight, p.o., to treat Cebus monkeys experimentally infected with about 200 cercariae of Schistosoma mansoni (SJ strain), via transcutaneous route. The oograms performed with rectal snips, as well as stool examinations carried out periodically, showed no viable eggs of the parasite, from day 29 to 226 post-treatment. The perfusion undertaken after killing the animals showed absence of worms in the treated monkeys, whereas 83 worms were recovered from the control, thus corroborating the results obtained by means of oograms and coproscopy. These results confirm the efficacy of 9-acridanone-hydrazones previously tested against the LE strain of S. mansoni. The low curative dose and apparent absence of toxicity render these drugs an important therapeutic reserve, taking into consideration the reports on the resistance of S. mansoni to the modern drugs oxamniquine and praziquantel.

Aspects of the granulomatous reaction in the liver of mice infected and reinfected with two different geographical strains of Schistosoma mansoni.

In this study, which was undertaken in relation to the histopathologic behavior of two different strains (LE-Belo Horizonte, MG and SJ-São José dos Campos, SP) in infections and reinfections (homologous or heterologous) with Schistosoma mansoni, the authors confirmed a more accentuated pathogenicity of the SJ strain. All the reinfections showed the presence of typical granulomas of the acute phase, when performed either with the same strain (homologous) or with a different strain (heterologous) of the parasite of the primo infection. The possible mechanisms responsible for reactivation of the immunopathologic response in reinfections are discussed.

Susceptibility of Biomphalaria tenagophila and Biomphalaria glabrata from a same region to 2 Schistosoma mansoni strains

B. tenagophila snails from Ouro Branco, MG, showed positivity for S. mansoni, with infection rates of 5%, 10%, (SJ strain), and 1% (LE strain) using a pool of miracidia. The mollusks were found to be susceptive from the 3rd generation reared in laboratory onwards. The B. tenagophila (OB, MG) when individually exposed to 10 miracidia, showed infection rate of 2% for LE strain. B. glabrata snails from Gagé, MG, showed a positivity rate of 58% for S. mansoni (LE strain), under experimental conditions. The B. tenagophila from Cabo Frio, RJ and B. glabrata from Belo Horizonte, MG used as a control for SJ strain showed infection rates of 47%-85% and 36% respectivily. For the LE strain, B. glabrata (BH, MG) used as control showed infection rate of 40%-75%.

Schistosoma mansoni: aspectos quantitativos da fertilidade e sobrevida de vermes oriundos de cercárias irradiadas com 3 Krad, em camundongos

The effect of gamma irradiation on the fertility of female mice, as well as the survival of worms in their portal system, have been observed in four groups of outbred albino mice (Mus musculus), experimentally infected with ca 450 cercariae of Schistosoma mansoni (LE and SJ strains), by transcutaneous route. The cercariae used were a) non-irradiated (control groups), and b) irradiated with 3 Krad of gamma irradiation (Co-60). From the 33rd day on, some stability in the population of surviving worm could be observed. This population remained constant till the end of the observation period (90th day), notedly in relation to the LE strain. Thus, it was concluded that gamma irradiation (at the dose of 3 Krad) is able to hinder the worm egg production in 98.1% of the infected mice. Further, it was observed that the few detected eggs were dead. Females were found to be more resistant to irradiation. The irradiation effect on the mortality of male worms was statistically significant scarcely from the 61st day on. The long period of permanence of the sterile adult irradiated worms in the portal system of mice and their probable involvement in the development of immunoprotection (the so-called concomitant immunity, without the immunopathological involvements for the host) are here discussed.

Schistosoma mansoni: evolução de vermes oriundos de cercárias irradiadas a nível de sistema porta, no camundongo

Eight groups of outbred albino mice were infected transcutaneously with cercariae of S. mansoni (strains LE and SJ) irradiated with either 3, 20 or 40 Krad, of gamma radiation from a cobalt-60 bomb and a control non irradiated group. Cercariae irradiated with 20 or 40 Krad failed to develop in the portal system and 3 Krad retarded development. Worms of the SJ strain developed more slowly than the LE strain.

Schistosoma mansoni: quantitative aspects of the development of irradiated cercariae in the skin, lungs and portal system, in mice

The migration of Schistosoma mansoni (LE and SJ strains) has been studied in eight groups of outbred Swiss albino mice (Mus musculus), which were previously infected with ca 450 cercariae, transcutaneously. The infection of mice was performed with non irradiated cercariae (control groups), or with gamma-irradiated cercariae, at the schedule of 3, 20 and 40 Krad. Regarding the skin, a progressive decrease was detected for the recovery rates, related to the time of infection. As far as the lungs and portal system are concerned, a significant inverse correlation was observed between the total recovery rate and the irradiation dosages. The dose of 20 Krad practically hinders the migration of the parasites (in both strains) from the lungs to the portal system, whereas the dose of 40 Krad prevents the migration of most of the parasites from the skin to the lungs.

Size of hepatic granuloma produced by eggs of 2 geographic strains of Schistosoma mansoni, in mice

The authors have determined the mean diameter of granulomas in the liver of mice infected with cercariae from two different and well definite geographic strains of Schistosoma mansoni (LE, Belo Horizonte, MG, and SJ, São José dos Campos, SP). A total of 1,170 granulomas has been measured. Granulomas measured on the 60th day after infection showed larger size than the other ones measured on the 90th day. Modulation of the immunopathologic response was significantly more efficient for the LE strain, whereas the granulomas (with 60 and 90 days) related to SJ strain were significantly larger. Data suggested a higher pathogenicity for the SJ strain. It is speculated whether these findings could explain, in part, the occurrence of regional variations of the anatomo-clinical forms of schistosomiasis.