Plasma calcitonin gene-related peptide in diagnosing and predicting paediatric migraine.

To investigate the role of plasma calcitonin gene-related peptide (CGRP) in paediatric migraine, we prospectively collected 134 blood samples during or between attacks from 66 migraine, 33 non-migraine headache (non-migraine) and 22 non-headache patients, aged 4-18 years. Plasma CGRP concentrations were measured by enzyme-linked immunosorbent assay and disability by Pediatric MIgraine Disability ASsessment (PedMIDAS) questionnaire. Migraineurs had higher plasma CGRP levels than non-migraine patients (P = 0.007). The attack level was higher than the non-attack level in migraine (P = 0.036), but not in non-migraine, patients. This was also revealed in paired comparison (n = 9, P = 0.015 vs. n = 4, P = 0.47). Using a threshold of 55.1 pg/ml, the sensitivity of the attack level in predicting migraine was 0.81, and specificity 0.75. The PedMIDAS score tended to be higher in the high CGRP (> 200 pg/ml, n = 7) group than in the low (< 200 pg/ml, n = 33) group (26.07 vs. 19.32, P = 0.16) using Mann-Whitney test. Plasma CGRP is useful for diagnosis in paediatric migraine.

Diagnosis of schistosomiasis using recombinant fructose-1,6-bisphosphate aldolase from a Formosan strain of Schistosoma japonicum.

Schistosoma japonicum obtained from Taiwan is a zoophilic strain that only infects domestic and small animals. Recombinant fructose-1,6-bisphosphate aldolase (FBPA) derived from this strain was used as an antigen in an enzyme-linked immunosorbent assay (ELISA) for the diagnosis of human schistosomiasis. The full-length DNA sequence of FBPA was found to be 1092 bp, encoding a protein of 363 amino acid residues, with a molecular mass of 39.6 kDa. A total of 120 participants were recruited from China and Taiwan to evaluate the diagnostic value of this recombinant protein. In these participants, 34 were found to be infected with S. japonicum, 16 with Ascaris lumbricoides, 15 with hookworm, 13 with Paragonimus westermani and 13 with Clonorchis sinensis, whereas 29 had no ova on faecal examination. Western blot analysis showed that the recombinant FBPA reacts strongly with schistosome ova-positive sera. The sensitivity and specificity of ELISA with FBPA were found to be 85.3% and 93.0%, respectively. These results indicate that FBPA derived from the Formosan strain of S. japonicum can be used for the diagnosis of human schistosomiasis.

Study of the morphological characteristics of Schistosoma mansoni in mice, squirrels, and hamsters.

This study attempted to describe mophological characteristics of S. mansoni worms. In the present study, 6 hamsters, 35 squirrels and 141 mice were infected with pooled cercariae of Schistosoma mansoni by intraperitoneal and percutaneous routes. The worm recovery rate was 18.2% (257/1,412) in hamsters (Mesocricetus auratus); 10.1% (3,310/32,792) in squirrels (Callosciurus erythraeus roberti) and 16.2% (4,328/26,720) in mice (Swiss strain). There were no significant differences between the sizes of 332 adults studied from three kinds of experimental animals. However, the worms collected from the hepatic portal system were usually larger than those from the peritoneal cavity because the latter almost always remained in the immature stage. We found male S. mansoni with tandem (17-22%), non-tandem (80-83%) and unusual/irregular arrangement (3-5%) of testes. The number of S. mansoni testes found were from 3 to 15 in mice, 3 to 11 in hamsters and 4 to 15 in squirrels. Mature worms had a tendency to reduce their size with aging. The number of S. mansoni cecal loops were from 1 to 5 in mice and hamsters and 1 to 4 in squirrels. The location of the first cecal loop was usually in the anterior part of body (1/4 in females and 1/3 in males), but there were some exceptions. The number of eggs in the uterus of each female worm, were 0-3 in mice, 0-1 in squirrels and hamsters. The average number was 0.75. The location of the ovary was usually situated in the anterior part of body of the worm in the three kinds of experimental animals. A few mated male and female worms of S. mansoni being free in the peritoneal cavity were found to develop to sexual maturity, because eggs were observed in their uteri. Their size was usually found to be considerably smaller than the worms seen in the hepatic portal system, and they had no hematin in their intestinal ceca. Encapsulated eggs were found from the peritoneal cavities of a few mice following intraperitoneal and percutaneous methods of infection.

Infectivity and development of the human strain of Hymenolepis nana in ICR mice.

In order to study the infectivity and development of the human strain of Hymenolepis nana in mice, a human strain of H. nana was inoculated into ICR mice. H. nana eggs were concentrated by the sedimentation method and inoculated by a disposable syringe (1 ml) connected to a long needle (8 cm) into the stomach of mice. Mouse feces were examined daily beginning day 5 after inoculation and the mice were sacrificed from days 19 to 65 post-infection (PI). The infection rate and worm recovery rate were 69% and 17%, respectively. The prepatent period ranged from 7 to 23 days. Autoinfection was found to occur in an ICR mouse infected with 60 eggs; 102 worms were recovered from its small intestinal lumen on day 19 PI. One row of hooklets was found on the scolex and the mean number of hooks was 19. The average length, width, and number of segments were 51 mm, 0.6 mm, and 1,099, respectively. The mean length and number of immature segments were 9 mm and 414 segments, mature segments 20 mm and 390 segments, and gravid segments 22 mm and 295 segments. The average length, width, and number of segments in 26 autoinfected worms were 11.5 mm, 0.3 mm, and 189 segments. The mean length and number of immature segments were 3.9 mm and 41 segments, mature segments 4.4 mm and 65 segments, and gravid segments 3.2 mm and 83 segments, respectively.

Hybridization of Schistosoma mansoni and Schistosoma japonicum in mice.

Crossing experiments in mice with two human species of Schistosoma japonicum (Taiwan strain) and Schistosoma mansoni (Puerto Rican strain) were performed. The hybrid miracidia from the cross between female S. japonicum and male S. mansoni infected both Biophalaria glabrata and Oncomalania h. chiui. However, those from the reciprocal crossing could infect only B. glabrata. B. glabrata infected with hybrid miracidia of female S. mansoni x male S. japonicum survived up to 30 days while those infected with miracidia of S. mansoni remained alive for more than 100 days after the first shedding of cercariae. Relatively few hybrid eggs reached maturity either in tissues or in the feces of infected mice. A low percentage of F1 eggs hatched and the infectivity of F1 miracida was also low. Morphology and behavior of hybrid eggs, miracidia, cercariae, and adults were similar to the maternal species. The daily egg production of the hybrid worm pair was less than that of the normal one. The observations in the present study may be attributed to the maternal effects. However, the phenomenon of parthenogenesis in schistosomes cannot be confirmed.

Viability of metacercariae of Clonorchis sinensis in frozen or salted freshwater fish.

Metacercariae of Clonorchis sinensis were collected from freshwater fish (Pseudorasbora parva) after storage in a freezer at -12 degrees C for 10-20 days, in a freezer at -20 degrees for 3-7 days, or treating with heavy salt (fish/salt = 10 gm/3 gm) at 26 degrees for 5-15 days. The metacercariae from the frozen fish were inoculated into 30 rats and eight rabbits, and those from the salted fish into 33 rats. Metacercariae from freshwater fish stored at -12 degrees for 10-18 days or -20 degrees for 3-7 remained viable and infective. Moreover, those kept at the heavy salt concentration for 5-7 days remained viable and infective. These findings indicate that freezing or storing infected freshwater fish in heavy salt may not be effective in the prevention of clonorchiasis. However, it appears that refrigeration, or keeping the fish in salt for longer periods, may be suitable for prevention of the infection.

Experimental infection of Philippine Taenia in domestic animals.

In the present study, six 34-44-day-old Small-Ear-Miniature pigs and one 14-day-old Holstein calf were each fed 10,000 Philippine Taenia eggs and sacrificed 27-43 days after inoculation. The infection rate was 100% for both pigs and calf with cysticerci recovery rates of 11 and 6%, respectively. A total of 6431 cysticerci were recovered only from the livers of the six pigs and 597 only from the liver of the calf; more occurred in the parenchyma (pigs 75%, calf 83%) than on the surface (pigs 25%, calf 17%). Mature cysticerci were found in four of the six pigs. A total of 317 cysticerci recovered from the pig livers were mature and the rest were either immature (926), degenerate or calcified (5188). All 597 cysticerci recovered from the liver of the calf were degenerate or calcified. Measurements of length, width, diameter of protoscolex, rostellum, and sucker and hooklet pattern indicated that Philippine Taenia is very similar to Taenia from Taiwan, Korea, Indonesia and Thailand and very different from classical T. saginata and T. solium.

Experimental infection of Indonesia Taenia (Samosir strain) in domestic animals.

Three 58-day old Small-Ear-Miniature (SEM) pigs, six 36-day old Landrace-Small-Ear-Miniature (L-SEM) pigs, and two 5-day old Holstein calves were each fed 3000 or 30,000 Indonesia Taenia (Samosir strain) eggs and sacrificed 27-195 days after inoculation. A total of 4922 cysticerci were recovered only from the livers of the three SEM pigs (1977 cysticerci) and six L-SEM pigs (2945 cysticerci). The infection rate in pigs was 100%. Cysticerci recovery rates of SEM and L-SEM pigs were 22.0% and 1.6%, respectively. Calves were not susceptible to Indonesia Taenia. More cysticerci were found in the liver parenchyma (L-SEM, 66.4%; SEM, 76.2%) than on the liver surface (L-SEM, 33.6%; SEM, 23.8%) of the infected animals. Most (99.86%) of the cysticerci recovered from the livers of L-SEM pigs were degenerated or calcified but 77.9% of those in the livers of SEM pigs were mature and only 22.1% were degenerated or calcified. Measurements of length, width, diameters of protoscolex, rostellum, and sucker and hooklet pattern indicated that Indonesia Taenia is very similar to Taiwan Taenia and very different from Taenia saginata and Taenia solium. The present findings indicate that Indonesia Taenia and Taiwan Taenia may be the same new species.

Experimental infection of Thailand Taenia (Chiengmai strain) in domestic animals.

Twenty-five gravid proglottides of a Thailand Taenia were obtained from a patient in Chiengmai, Thailand, and brought to our laboratory. The tapeworm was determined to be T. saginata-like by counting uterine branches (mean number 16, range 12-19 on each side). The eggs from these proglottides remained infective under storage at room temperature for 14 days followed by refrigeration (4-8 degrees C) for 131 days. Eight Small-Ear-Miniature pigs and two Holstein calves were each fed with 3000 eggs and sacrificed 12-76 days afterwards. Six pigs became infected and 16 cysticerci were recovered from the livers. Thirteen degenerated/calcified cysticerci were also recovered from the livers of the two calves. More cysticerci were found in the liver parenchyma (55%) than on the liver surface (45%) of the infected animals. Measurements of length, width, diameters of protoscolex, rostellum and sucker and hooklet pattern show that Thailand Taenia is similar to Taenia from Taiwan, Korea and Indonesia but different from T. saginata and T. solium. These findings indicate that Thailand Taenia, Taiwan Taenia, Korea Taenia, and Indonesia Taenia may be of the same species or sub-species.

Poor efficacy of albendazole for the treatment of human taeniasis.

In order to evaluate the efficacy of albendazole for the treatment of taeniasis, regimens of 400 mg x 1 day, 800 mg x 2 days, 800 mg x 3 days, 1200 mg x 2 days, and 1200 mg x 3 days were compared. Of 66 cases treated and investigated 7-14 days after treatment, 52 were still expelling proglottids. Three months posttreatment, these cases were re-treated with atabrine at 1.2 g per case for males and 1.0 g per case for females. Fifty-seven patients expelled worms or parts of tapeworms. The nine negatives may represent the number cured by the treatment with albendazole. The cure rates with albendazole for various regimens were up to 50% for 800 mg x 3 days, 1200 mg x 2 days or 1200 mg x 3 days, 14.3% for 800 mg x 2 days, and 0% for 400 mg x 1 day or 800 mg x 1 day. This study shows that albendazole is not very effective in the treatment for taeniasis.

Experimental studies on the pathway for migration and the development of Taiwan Taenia in domestic pigs.

The pig is the most favorable experimental intermediate host of Taiwan Taenia. Cysticerci in infected pigs are located in the liver except for a few extrahepatic ones. The present study was designed to investigate the pathway of migration of the oncospheres of Taiwan Taenia in the pig. In the first group, each of 5 Small-Ear-Miniature (SEM) and one Landrace-Small-Ear-Miniature (L-SEM) pigs were injected with 5000 hatched oncospheres into the ear vein. Three SEM and one L-SEM pigs were found to harbor 88 degenerated or calcified cysticerci only in the liver 51-81 days after injection. In the second group, each of 3 L-SEM pigs were injected with 5000 hatched oncospheres into the jugular vein. One of the 3 pigs was found to have 5 cysticerci (2 mature and 3 degenerated or calcified) only in the liver at 89 days post-injection. In the third group, each of 4 SEM, three L-SEM, and 3 L-SEM pigs were injected with 10,000, 10,000, and 5000 hatched oncospheres, respectively, directly into the portal vein after surgical opening of the abdominal cavity. All 10 pigs were found to be infected with a total of 1088 cysticerci (44 mature and 1044 degenerated or calcified) only in the liver 23-62 days after injection. Although the sites of injection in these three groups were different, the liver was the only final location of the cysticerci. These findings give strong evidence that the oncospheres migrate to the liver through the venous circulation and develop in this organ.

A comparative study on egg hatching methods and oncosphere viability determination for Taenia solium eggs.

The efficiencies of an enzyme method and the sodium hypochlorite method in hatching eggs of Taenia solium (Henan strain) were compared. The viability of the hatched oncospheres was determined by staining with trypan blue. For the former method, the hatching rate and viability percentage were 85.7% and 79.7%, respectively. The corresponding figures for the latter method were 97.9% and 86.7%, respectively. These findings indicate that the sodium hypochlorite method is a simple, efficient and practical method for hatching eggs of Taenia solium.

In vitro hatched oncospheres of Asian Taenia from Korea and Taiwan develop into cysticerci in the peritoneal cavity of female scid (severe combined immunodeficiency) mice.

In vitro hatched (but not activated) oncospheres of Asian Taenia obtained in Korea and Taiwan, prepared by the sodium hypochlorite method, rinsed with sterile PBS several times and adjusted to 5 x 10(4)/0.5 ml PBS, were injected intraperitoneally or subcutaneously into male or female scid mice of 3 different strains. When these scid mice were sacrificed 4 months later, the females harboured fully developed cysticerci either in the peritoneal cavity or under the back skin, whereas males did not. All cysticerci from the peritoneal cavity were easily recovered by rinsing the abdomen with PBS. Although most cysticerci recovered from pig liver usually become calcified within 1-2 months, in female scid mice they all increased in size and were viable. Intraperitoneal (i.p.) injection of in vitro hatched oncospheres is recommended for easier recovery of Asian Taenia metacestodes in laboratory animals.

Filariasis eradication on Kinmen Proper, Kinmen (Quemoy) Islands, Republic of China.

A control program for bancroftian filariasis was conducted on Kinmen Proper from 1970 to 1982. The combined method of mass chemotherapy with diethylcarbamazine (DEC) for microfilarial carriers and larvicide with sumithion for mosquitoes was used on the island for 3 years (1974-76). The microfilarial rate and microfilarial density decreased from 8.4% and 14.6/20 microns in 1970-73 to 0.7% and 5.3/20 microns in 1976. The infection rate, infective rate, and larval density of Culex quinquefasciatus, the only mosquito vector of the parasite, decreased from 10.3%, 6.4%, and 6.4 in 1970-73 to 0.3%, 0.2%, and 1.3, in 1976, respectively. For final elimination of the infection, DEC medicated salt (0.33%, w/w) was then administered to the whole population for 6 months (March-September 1977). All accepted the program and no adverse side-effects were noted. Six annual follow-up surveys (1977-82) revealed only 3 out of 3853 previous carriers to be positive. These carriers were cured with a course of 5 g DEC. All family members of the 3 carriers and of the 12 neighbouring families were also given DEC medicated salt for 4 months in order to clear the only remaining source of filarial transmission. In conclusion, administration of DEC medicated salt is simple, rapid, safe, inexpensive, efficient, and practical for filariasis control or eradication.

Eating habits of east Asian people and transmission of taeniasis.

In order to understand the role of raw meat and viscera eating habits in the transmission of taeniasis in Asian countries, 1502 infected aborigines in ten mountainous districts/towns of six counties in Taiwan, 58 infected persons in two villages on Cheju Island, Korea, and 97 cases in Ambarita District on Samosir Island, North Sumatra, Indonesia were studied during the field surveys. All infected Taiwan aborigines had the habit of eating raw meat and viscera of wild and/or domestic animals. Among these aborigines, 73% ate wild boar, 66% flying squirrel, 65% wild goat, 56% muntjac, 49% wild rats, 46% monkey, 38% hare, 20% civet-cats, 18% weasel, 17% pheasant, 14% squirrel, 4% grouse, 1% deer, 1% snake, less than 1% bamboo partridge, less than 1% frog, less than 1% bear, less than 1% dog, and less than 1% fox. Of the 58 infected persons with Taenia on Cheju Island, Korea, 72% ate raw meat and/or viscera of pig and cattle, 19% raw pork only, and 9% raw beef only. Among 12 infected persons infected with T. saginata-like tapeworms, 7 had eaten raw pork, 2 raw beef and pork and 3 raw pork. Almost all of the 97 cases of taeniasis on Samosir Island of North Sumatra, Indonesia, had eaten only undercooked pork. Eleven of 15 cases were found to be infected with T. saginata-like tapeworms. Eating habits observed suggest an unusual way of transmission of Taenia in East Asia.

Review of enterobiasis in Taiwan and offshore islands.

From 1995 to 1997, we carried out extensive surveys of Enterobius vermicularis infection among school children in Taiwan and offshore islands. Among 6,315 children at 80 schools in 21 towns and districts of 8 counties tested, an overall infection rate was 11%. The pre-school children had the highest rate of 14%. These findings indicate that pinworm infection remains highly prevalent in Taiwan and offshore islands. Moreover, in this paper, the results from the studies conducted by parasitologists of Taiwan in the past 43 (1955-1997) years were reviewed. This information included epidemiology, diagnosis, chemotherapy, and control methods, may provide some useful suggestions on the prevention of pinworm infection specialists and agencies concerned with the control of this disease.

Taenia saginata asiatica: epidemiology, infection, immunological and molecular studies.

The taeniasis in East Asia has a special epidemiological pattern: people eat meat and/or viscera of pigs and acquire infection of Taenia saginata-like tapeworms. However, cysticercosis is more often found in pigs than cattles. In order to elucidate the taxonomic status of this parasite, we have conducted extensive field surveys, experimental infections, and morphological as well as immunological studies since 1981. After obtaining sufficient information from our studies, we finally came to the conclusion that the T. saginata-like tapeworm in Asia is a new subspecies of T. saginata and was named as T. saginata asiatica. The classical T. saginata was renamed as T. saginata saginata. In this paper, we provide the history of T. s. asiatica in Asia.

Experimental studies on physiological and morphological aspects of Cysticercus cellulosae in pigs.

Three Small-Ear-Miniature, 3 Landrace-Small-Ear-Miniature, and one Douc-Yorkshire-Landrace pigs were inoculated orally with 100 000 eggs of Zhengzhou strain or 10 000 eggs of Harbin strain of Taenia solium. A total of 3739 cysticerci were recovered from 3 Small-Ear-Miniature and 3 Landrace-Small-Ear-Miniature pigs, giving an infection rate of 85.7% and a cysticercus recovery rate of 1.1%. The predilection sites of Cysticercus cellulosae in descending order were leg muscles, abdominal muscles, thoracic muscles, liver, head muscles, diaphragm, tongue, heart, trachea, and omentum/testes. Except 2 calcified cysticerci in the tongue, 2 in the heart, and 176 in the liver, the remaining cysticerci were all alive. The greatest number of cysticerci per 100 g of muscles or viscera was found in the head muscles, followed by the leg, diaphragm, heart, tongue, thoracic, abdominal, omentum, testes, and trachea. All cysticerci were evaginated in pig's bile after fluid was drawn out from cysticerci, whereas evagination occurred in only 83.2% of those without fluid drawing. In 364 evaginated cysticerci, the mean length and width of scolex, proglottid, and bladder, and diameter of rostellum and sucker were 826 x 747 microm, 5,370 x 1,734 microm, 2,885 x 3,002 microm, 155 microm, and 253 microm, respectively. In the protoscolex, the mean number of segments was 33. Each cysticercus had 2 rows of rostellar hooks on the scolex, and the mean length and width of inner and outer hooks were 151 x 18 microm and 117 x 14 microm, respectively. The number of paired hooks ranged from 10 to 18.

The serological response of pigs experimentally infected with a species of Taenia from Taiwan.

An enzyme-linked immunosorbent assay (ELISA) for the detection of antibody in pigs infected with a possibly new species of Taenia isolated in Taiwan is described. The test antigen ThFAS was fractionated from the cyst fluid of a heterologous cestode Taenia hydatigena. In lightly infected pigs (less than or equal to 4 recovered cysts at necropsy 17 weeks post-inoculation), antibody was detected as early as 3 weeks post-inoculation. In more heavily infected pigs (6-72 recovered cysts at necropsy 32 weeks post-inoculation), antibody was still detectable at the time of necropsy. Cysticerci were found only in the livers of the infected pigs. This ELISA should be highly useful for detecting infection of pigs with this larval cestode in regions where the presence of Taenia solium is unlikely.

Egg production capacity of Paragonimus pulmonalis in cats.

In the present study, each of 43 cats was fed with a single metacercaria of Paragonimus pulmonalis and 2 cats (controls) with 2 metacercariae. They were killed 22-372 days postinfection. In the test group, 18 (41.9%) cats were infected, as well as the 2 cats in the control group. Among the 18 worms recovered from the test group, 9 found in the lungs were mature, whereas 5 mature and 4 immature worms were in the pleural cavity. Most eggs from worms encased in a cyst were passed in the feces. However, eggs from worms in the pleural cavity were only found on the lung surface and pleural cavity. Mature worms in cysts seemed to exhibit higher egg production capacity than those in the pleural cavity. Paired mature worms from a cyst laid more eggs than a single encysted mature worm in vitro at room temperature.