Novel real-time PCr for detection of Schistosoma japonicum in stool.

Chemotherapy has been used on a large scale in countries where the blood fluke Schistosoma japonicum is endemic. This has led to a lower intensity of infections and consequently lower diagnostic values of commonly used diagnostic tests like serology and Kato-Katz stool smear. We designed a novel real-time PCR method for detection of S. japonicum in stool samples. Further, we evaluated different versions of an inexpensive, non-commercial extraction method, ROSE, as well as the commercial QIAamp DNA Stool Mini Kit. PCR primer sequences were designed targeting the mitochondrial NADH dehydrogenase I gene. Bovine serum albumin was added to the DNA extracts and SYBR Green was used for detection. The PCR method was evaluated with non-infected stool samples spiked with S. japonicum eggs. It demonstrated high sensitivity, even in samples containing a single egg. The two extraction methods were equally effective. The PCR was specific for S. japonicum when tested against other Schistosoma species, Trichuris trichiura, hookworm and Taenia sp. We conclude that this novel real-time PCR, in combination with either ROSE or QIAamp DNA Stool Mini Kit extraction, is a sensitive and specific tool for diagnosing S. japonicum in human stool samples.

An epidemiological investigation of congenital Schistosoma japonicum transmission in Hubei Province, PR China.

A cohort study was conducted in Hubei Province, China, following serious flooding of the Yangtze River in the autumn of 1998 to investigate the possibility of congenital transmission of Schistosoma japonicum in humans. The cohort investigated was comprised of 205 women and their 208 infants born between 1 September and 30 December 1998. Blood and fecal samples from all the women and their infants were collected and examined for S. japonicum infection. Positive specific antibody titers were found in 14 (6.8%) of the mothers, but no fecal egg excretion was observed. All infants had negative specific antibody titers and no S. japonicum eggs were found in their feces. Hence, the present study coud not confirm congenital S. japonicum transmission in humans. Further studies are highly wanted to study the impact of prenatal exposure of S. japonicum on the offspring.

Schistosoma japonicum in the pig: a new technique for estimation of intestinal tissue egg counts.

This study introduced a new method for estimating intestinal tissue Schistosoma japonicum egg counts, based on scraping of the mucosal layer of different sections of the intestines. Twenty-eight Danish Landrace/Yorkshire/Duroc crossbred pigs were divided into 3 groups of 15, 5 and 8 pigs, respectively. Pigs were fed either a high- or low- protein diet and were infected by an intra-muscular or per-oral route of infection with doses of either 1,000, 1,500 or 3,000 S. japonicum cercariae. The pigs were killed 9-11 weeks post infection. For all 28 pigs the intestines were divided into 3 sections: cecum, colon and rectum and the entire mucosa was scraped off the serosa of each section and homogenized. Subsequently, samples corresponding to 5 g homogenised mucosal tissue were digested and egg counts were determined and correlated to liver egg counts. In order to compare the relative distribution of eggs in the mucosa and the serosa, small intestinal wall subsamples formerly taken from each section from a subgroup of 5 pigs were homogenized and egg counts determined for both the mucosa and serosa. The number of eggs were significantly higher in the mucosa than in the serosa. Egg counts estimated from digestion of mucosa subsamples either over or underestimated egg counts based on scrapings of the entire mucosa when compared, reflecting the very patchy distribution of S. japonicum eggs in the intestinal wall. Correlating liver egg counts with the number of eggs based on scrapings from the entire mucosa from cecum, colon and rectum, respectively, significant correlations were found for 2 out of 3 groups of pigs. The present study revealed that estimating intestinal tissue egg counts based on scrapings of the entire mucosa is a reliable and convenient approach, nicely supporting the liver tissue digestion approach. In addition, a reduction of the processing time of intestinal tissue in general was achieved due to the very simple scraping technique.

Schistosoma japonicum in the pig: the influence of age on the host/parasite relationship.

The current study sought to elucidate a possible association between age and susceptibility to a primary infection with Schistosoma japonicum in pigs. Sixteen Landrace/Yorkshire crossbred specific pathogen-free pigs in three different age groups (group A-C), aged approximately 7, 24 and 37 weeks at the beginning of the experiment, were infected by intramuscular injections of 1,000, 1,500 or 2,400 cercariae, respectively. Fecal egg counts were obtained twice weekly from six to eight weeks post infection (wpi), and the pigs were killed 11 wpi. The number of worms collected were counted and sexed subsequent to perfusion. Tissue egg counts were estimated on samples from the liver. The worm recoveries for group A, B and C were 3.2%, 8.1% and 3.8%, respectively. No differences were observed between the male/female ratios of the three groups. The fecundity parameters, ie, fecal egg counts per mature female and liver egg counts per mature female, showed no significant differences between the three age groups. The results did not indicate any difference in susceptibility between the different age-groups of pigs to a primary infection with S. japonicum.

A new technic for counting Schistosoma japonicum eggs in pig feces.

An improved laboratory method was developed for counting Schistosoma japonicum eggs in pig feces, which involves filtration, sedimentation and centrifugation, but avoids toxic chemicals. It is sensitive, allows easy differentiation from similar-sized and shaped protozoan cysts, and permits evaluation of egg viability both by direct viewing of eggs and miracidial hatching. It was found to be significantly better at recovering eggs than the modified Bell filtration technic. The sensitivity, specificity and practicality of this technic make it our method of choice for studies on porcine schistosomiasis japonica.

The population dynamics of cercariae of Schistosoma japonicum in Oncomelania hupensis.

The population dynamics and production of cercariae of Schistosoma japonicum in Oncomelania hupensis are reported. The experiments covered the whole life span of positive snails and different intervals of cercariae shedding. The results indicated that two patterns of the dynamics of cercariae shedding had been found in the life span of positive snails. The first was a long-time interval (4-7 days) and progressive decline pattern. The cercariae shedding of positive snails lasted 18-19 weeks in males and for 32-33 weeks (once a week). The second was a short-time interval (1-3 days) and continued release pattern. The cercariae shedding of positive snails lasted for 20-36 days (every day shedding). Shedding cercariae stimulate cercariae development.