Enterobius vermicularis infection in schoolchildren: a large-scale survey 6 years after a population-based control.

Pinworm infection remains prevalent in children in many parts of the world. This study was designed to determine the prevalence of this infection in schoolchildren in Taiwan after the termination of the 15-year population-based control project in 2001. Our results showed that 2.4% of 118 190 children in 385 primary schools were found to have enterobiasis by two-consecutive-day adhesive cellophane perianal swabs. The prevalences were significantly different in the 25 counties/cities surveyed (0.6-6.6%). A significantly higher prevalence was found in boys (2.6%) than in girls (2.2%) and the prevalence decreased by grade from 3.8% in grade 1 to 1.0% in grade 6. In the primary schools, 9.1% had positive rates 10%. In addition, pinworm infection was found to be significantly associated with the socioeconomic status, personal hygiene and sanitary conditions of the children. The results indicate that the overall prevalence of enterobiasis remains at a low level after the control programme was transferred to the local governments.

Detection of Kobe-type Babesia microti associated with Japanese human babesiosis in field rodents in central Taiwan and southeastern mainland China.

Field rodent surveys for Babesia infection were performed from 2002 to 2005 in the vicinities of human babesiosis occurrences in Taiwan and mainland China. Babesia microti was identified by microscopical examination and/or PCR in 1 Rattus coxinga and 1 Crocidura horsfieldii in central Taiwan and in 13 Niviventer confucianus and 1 Apodemus agrarius in Zhejiang and Fujian Provinces of southeastern China. Of 15 B. microti samples detected by PCR, all except 1 were shown to be the Kobe-type, the aetiological small subunit rRNA gene-type of the first Japanese patient; the exception was also a Kobe-related type. The Kobe-type had been found in rodents only in a few places including the human infection occurrence place in Japan. The internal transcribed spacer 1 to 2 sequences of the Taiwanese and Chinese Kobe-types were very similar to each other but considerably different (approx. 94% pairwise identities) from that of the Japanese Kobe-type. A Taiwanese Kobe-type strain was serologically differentiated from the Kobe strain originating from the Japanese first patient. The distribution of the Kobe-type in the vicinities of human babesiosis occurrences in Taiwan and China as well as in Japan is suggestive of involvement of the Kobe-type in Asian human babesiosis.

Cockroaches (Periplaneta americana and Blattella germanica) as potential mechanical disseminators of Entamoeba histolytica.

Since cockroaches feed on feces, they may disseminate infective agents with the fecal-oral route. A field survey and experiments were conducted to determine the role of cockroaches in the dissemination of Entamoeba histolytica. Periplaneta americana (n=299) and Blattella germanica (n=29) were collected from 11 primary schools in an urban area of South Taiwan. E. histolytica/E. dispar cysts were found on the cuticle and/or in the digestive tract of P. americana (25.4%) whereas cysts were only isolated from the digestive tract of B. germanica (10.3%). E. histolytica cysts were found on the cuticle and/or in the digestive tract of both species after exposure to 1000 or 100 cysts/g feces for 24 h. These findings indicate that cockroaches may harbor the E. histolytica cysts and play a role as potential mechanical disseminators.

Current status of human parasitic infections in Taiwan.

The eradication of the 2 mosquito-borne parasitic diseases, malaria and lymphatic filariasis, is one of the greatest achievements of the parasite control campaigns in Taiwan. Most of the soil-transmitted nematode infections, with the exception of pinworm infection, are currently well controlled and limited to some aboriginal areas. Food-borne parasitic zoonosis such as infections with Angiostrongylus cantonensis, Clonorchis sinensis, and Taenia saginata asiatica are not rare, but the former is seasonal and the latter 2 are ethnically and geographically associated. Intestinal protozoal infections with Giardia lamblia and Cryptosporidium parvum are at low levels but may be widely distributed. Opportunistic protozoal infections among patients with acquired immunodeficiency syndrome, which included amebic colitis, Pneumocystis carinii pneumonia, and cerebral toxoplasmosis, are becoming increasingly important. The rapid increase in international travel and the introduction of large numbers of foreign workers from other countries in Southeast Asia may change the epidemiological patterns of parasitic infections in Taiwan.

The growth effects of gamma-ray irradiation on third-stage larvae of Angiostrongylus cantonensis in snail.

In order to clarify growth effects of gamma-ray irradiation on the third-stage larvae of Angiostrongylus cantonensis, snails naturally infected with larvae of A. cantonensis were irradiated with a cobalt-60 source, and the total dosage of exposure was 0.1 kGy. Third-stage larvae were collected from each group of irradiated and non-irradiated snails and then administered to rats. A significant reduction was observed in the number of recovery worms from rats sacrificed at 45 days after infection compared with the control group. Moreover, there was an increase in the percentage of female worms, and the body length of the irradiated group's worms was significantly shorter than that of the control group. In ultrastructural observations by transmission electron microscope, the cuticle was thickened and the subcuticle showed partial vacuolization and separation from muscle in the irradiated group. In addition, the muscles myofibril became shorter, the microvilli of intestinal mucosa were shorter and irregular, the terminal web disappeared, and the phagocytic vesicles, mitochondria, lysosomes and secretion granules decreased in number, respectively. Focusing on the mitochondria of the epithelial cells of the intestine, there were degenerative changes in the irradiated group showing vacuolization. The gonads of female worms recovered from the irradiated group revealed degenerative vacuolization, and the gonads of male worms could not be found.

Parasitic infections among the aborigines in Taiwan with special emphasis on Taeniasis asiatica.

According to the results obtained by our laboratory and the Department of Parasitology, Taipei Medical College, from 1995 to 1998, the overall infection rate of enterobiasis was 16% (4-29%) among 4,426 primary school students at 64 schools in 13 mountainous districts of 10 counties, and the infection rate of head louse infestation was 12% (3-26%) among 5,121 students. A continuous series of field studies on epidemiology and chemotherapy of taeniasis asiatica have been conducted between 1971 and 1992 among 27,359 aborigines in 88 villages of 14 districts in 10 counties and 3,104 were found to be infected, giving an overall infection rate of 11%. Laboratory studies have been performed on the morphology, intermediate hosts, experimental infection, immunology and molecular biology. Moreover, development of Taiwan Taenia in one of 3 volunteers has been successfully demonstrated since 1988. Moreover, the domestic pig was identified as the intermediate host of Taiwan Taenia as in the classical T. saginata (Poland strain). Most of cysticerci were developed in or on the parenchyma of liver. The results indicated that Taiwan Taenia is different from classical T. saginata. It has been named as T. s. asiatica. Moreover, the classical T. saginata was renamed as T. s. saginata. This paper summarizes findings and reviews those reported by our laboratory. In addition, the clinical manifestations and eating habits of the aborigines are also discussed.

Current status of imported parasitic infection among foreign workers in northern Taiwan (1999-2000).

In the present study, a simple, economic and practical technique was employed for stool examination. Of a total of 6,146 fecal samples from foreign workers in Northern Taiwan between 1999 and 2000 were examined, 615 were found to be positive for parasitic infection and the overall infection rate was 10%. Newly arriving foreign workers had a significantly higher infection rate (15%) than those who had worked in Taiwan for 6-12 months (8%). The foreign workers came from Indonesia, Philippines, Thailand, Malaysia, and Vietnam. Except for the small number of workers from Malaysia which was not included, the infection rate order by nationality was Vietnamese (21%) > Indonesian (13%) > Philippino (10%) > Thai (4%). The female examined workers were about 3-fold of males and their infection rate (11%) was also significantly higher than the males (5%). The order of rates by age was 20-30 years (11%) > 31-40 years (8%) > 41-50 years (5%). According to the species of parasites, 569 foreign workers were infected with 1 species (9%) > with 2 species (0.7%) > with 3 species (0.1%). Totally, 14 species (10 helminths including 1 plant nematode, Heterodera and 4 protozoa; hookworm might include 2 or 3 species, but counted as one species here) were found, of which 10 species were pathogenic (9 helminths and 1 protozoa) and 4 non-pathogenic. Foreign workers from Indonesia harbored 12 species of parasites > from the Philippines, 9 species > from Thailand, 8 species > from Vietnam, 7 species.

Capillaria hepatica: a spurious case with a brief review.

A 12-year-old Bunun aboriginal school student in Sanmin District Kaohsiung County, Southern Taiwan, was found to have Capillaria hepatica eggs in the feces during a survey on intestinal parasitic infection in 1997. Moreover, this student was also infected with Trichuris trichiura and Ascaris lumbricoides. The C. hepatica eggs were light yellowish-brown in color, 54 (40-60) microns in length and 29 (25-35) microns in width with a 3 (3-4) microns thick shell and two colorless egg plugs 7 (5-9) x 6 (5-8) microns. This is the eighth case of human spurious C. hepatica infection in Taiwan. In addition to the reporting of this case, the genuine and spurious infections of C. hepatica in humans as well as its infection in rats around the world are briefly reviewed.

A survey on the intestinal parasites of the school children in Kaohsiung county.

The present study on the prevalence of intestinal parasites infection, from September to December 1999, was conducted among school children in Taoyuan Hsiang, Kaohsiung county. The investigated areas included three (Jiannshan, Shingjong and Taoyuan) primary schools. The overall infection rate in 305 children determined by Merthiolate-Iodine-Formaldehyde Concentration method of stools was 17%. Four confirmed species of helminthes (Ascaris lumbricoides, Hookworm, Trichuris trichiura and Hymenolepis nana) and three species of protozoa (Giardia lamblia, Entamoeba coli and Blastocystis hominis) were detected. Males and females had the infection rates of 24% and 11%. The infection rates of aboriginal and non-aboriginal children were 17% and 14%, respectively. Grade 1 and Grade 6 had the highest infection rate (21%). Following tape perianal examination of 302 children, the overall infection rate of Enterobius vermicularis was 25%. Males and females had the infection rates of 24% and 26%. The infection rates of aboriginal and non-aboriginal children were 27% and 11%, respectively. Grade 1 had the highest infection rate (37%). Based on these data, the infection rate of intestinal parasites among rural primary school children in Kaohsiung county remains high.

An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group.

BACKGROUND: Enteroviruses can cause outbreaks of hand-foot-and-mouth disease (characterized by vesicular lesions on the hands, feet, and oral mucosa) or herpangina, usually without life-threatening manifestations. In 1998 an epidemic of enterovirus 71 infection caused hand-foot-and-mouth disease and herpangina in thousands of people in Taiwan, some of whom died. METHODS: We assessed the epidemiologic aspects of this outbreak. Cases of hand-foot-and-mouth disease or herpangina in ambulatory patients were reported to the Taiwan Department of Health by a mean of 818 sentinel physicians. Severe cases in hospitalized patients were reported by 40 medical centers and regional hospitals. Viruses were isolated by 10 hospital laboratories and the department of health. RESULTS: The sentinel physicians reported 129,106 cases of hand-foot-and-mouth disease or herpangina in two waves of the epidemic, which probably represents less than 10 percent of the estimated total number of cases. There were 405 patients with severe disease, most of whom were five years old or younger; severe disease was seen in all regions of the island. Complications included encephalitis, aseptic meningitis, pulmonary edema or hemorrhage, acute flaccid paralysis, and myocarditis. Seventy-eight patients died, 71 of whom (91 percent) were five years of age or younger. Of the patients who died, 65 (83 percent) had pulmonary edema or pulmonary hemorrhage. Among patients from whom a virus was isolated, enterovirus 71 was present in 48.7 percent of outpatients with uncomplicated hand-foot-and-mouth disease or herpangina, 75 percent of hospitalized patients who survived, and 92 percent of patients who died. CONCLUSIONS: Although several enteroviruses were circulating in Taiwan during the 1998 epidemic, enterovirus 71 infection was associated with most of the serious clinical manifestations and with nearly all the deaths. Most of those who died were young, and the majority died of pulmonary edema and pulmonary hemorrhage.

Improvements in the infectivity of cryopreserved third-stage larvae of Angiostrongylus cantonensis using a programmable freezer.

Although there have been some advances in the cryopreservation of Angiostrongylus cantonensis, the degrees of viability and infectivity of the cryopreserved developmental stages have not been high. A two-step freezing protocol using a programmable freezer was determined to be effective in improving the infectivity of the cryopreserved third-stage larvae of this parasite. After washing steps and suspension in 10% (v/v) dimethylsulfoxide and equilibrium at room temperature the larvae were placed into the freezer. The temperature was lowered first at 0.8 degrees C/min from room temperature to -40 degrees C and then at 10 degrees C/min to -70 degrees C. The samples were plunged into liquid nitrogen. After storage in liquid nitrogen for 7-15 days the larvae were thawed rapidly in 37 degrees C water and 27.6% were found to show vigorous "S-shape" movement without significant changes in appearance. These larvae (50/rodent) could develop to the fifth stage in mice (42.6%) and establish patent infection in rats (40.4%). Moreover, there was no significant difference in the recovery rates of cryopreserved worms and their unfrozen counterparts. These findings indicate that steady precooling conditions may decrease damage with regard to the infectivity of cryptopreserved third-stage larvae of A. cantonensis.

Humoral immune responses in mice infected with gamma-irradiated third-stage larvae of Angiostrongylus cantonensis.

Humoral immune responses in C57BL/6 strain mice infected with 1.0 kGy gamma-irradiated third-stage larvae (L3) and with non-gamma-irradiated L3 of Angiostrongylus cantonensis were assessed in this study. The young-adult worm (L5) antigen-specific antibodies in sera of both groups of mice elevated gradually and reached a peak in the third week after infection. The IgM antibodies of mice 1-week after infection with gamma-irradiated L3 and with non-gamma-irradiated L3 recognized several L5 antigens of the same molecular weight. However, only partially identical profiles were observed in the reaction of L5 antigens with IgM and IgG antibodies from mice 3 weeks after infection with gamma-irradiated L3 and with non-gamma-irradiated L3 respectively by western blot analysis. With regard to antibody-dependent cell-mediated cytotoxicity, antibodies from 3-week infected mice significantly increased the adherence of mouse eosinophils and neutrophils to L3 of A. cantonensis. The immunoresponses of splenic cells from mice in the third week of infection to sheep red blood cells were suppressed because the number of plaque forming cells obviously decreased more than the uninfected control.

Detection of circulating antigen by monoclonal antibodies for immunodiagnosis of angiostrongyliasis.

Two monoclonal antibodies, which recognize the epitope on an antigen with a molecular weight of 204 kD from the fifth-stage worm of Angiostrongylus cantonensis, were previously prepared and used to detect circulating antigens in patients with eosinophilic meningitis or meningoencephalitis and in mice experimentally infected with this parasite by a double-antibody, sandwich enzyme-linked immunosorbent assay (ELISA). The levels of this circulating antigen in experimentally infected mice were significantly higher three weeks after infection. The ELISA values in the detection of circulating antigens in cerebrospinal fluid (CSF) from patients were markedly higher than those in serum. Immunodiagnosis of patients with angiostrongyliasis by this technique proved to be highly specific for circulating antigens in serum and CSF specimens; however, the sensitivity in CSF was significantly higher than in serum.

Role of T cell subpopulations in mice infected with Angiostrongylus cantonensis.

When C57BL/6 mice were infected with Angiostrongylus cantonensis, the percentage of T helper (CD4+) cells and T supressor (CD8+) cells in peripheral blood increased weekly until the third and seventh week respectively, and then gradually decreased. C57BL/6 mice were depleted of CD4+ and CD8+ T cells by in vivo injection of anti-CD4 and anti-CD8 monoclonal antibodies, respectively, and then infected with A. cantonensis. There were significantly more and less worms recovered in the mice depleted of CD4+ and CD8+ T cells respectively than in undepleted mice. Discrete subpopulations of T cells from mice exposed to A. cantonensis for 3 weeks or 7 weeks were adoptively transferred to syngeneic recipients which were then given a challenge infection. Protection was mediated by a CD4+ T cell population present in mice after 3 weeks of infection but was not demonstrable with cells taken 7 weeks after infection. When CD4+ T cells obtained from 3-week infected mice were mixed with 5% CD8+ T cells obtained from mice infected for 7 weeks, no significant transfer of resistance was observed. Thus, immune responses to A. cantonensis in mice were regulated by discrete subpopulations of T lymphocytes.

[Parasitic infections among wild rats from two areas of Kaohsiung].

Wild rats including four strains were captured for examination of parasitic infections. The numbers were 101 from areas of Kaohsiung harbor and 98 from areas of Kaohsiung airport. Most of the examined wild rats were Rattus norvegicus and second most were Suncus murinus. The infection rates of intestinal helminths in rats of both areas did not display any significant difference. The overall infection rate was 29.6%. The infection rate was highest in R. norvegicus and the most frequently found parasite in rats was Hymenolepis diminuta. Moniliformis moniliformis and Angiostrongylus cantonensis were revealed in two separated R. norvegicus rats. The infection rates of intestinal helminths in R. norvegicus were significantly correlated with the size of rats. There was no find in microscopic observation of blood smear from these rats stained with Giemsa's dye.

The susceptibility of five stains mice to infections with Angiostrongylus cantonensis.

The susceptibility to the infection of Angiostrongylus cantonensis was studied in five inbred stains of mice including four H-2 haplotypes. Three weeks after infection, C57 BL/10J (H-2b) and C57BL/6J (H-2b) stains of mice showed the lowest, CBA (H-2k) and A/J (H-2a) stains showed intermediate, and the BALB/cJ (H-2d) mice showed the greatest susceptibility to infection. BALB/cJ mice also displayed the most body weight loss, the highest number of worms and showed the least number of days of survival. The s susceptibility of C57BL/6J (H-2b) and C57BL/10J (H-2b) strains of mice sharing same H-2 haplotype was the same despite their different genetic backgrounds. These data suggested that the H-2 gene might influence the susceptibility of mice to A. cantonensis.

[Investigation on the parasites of alien laborers in Kaohsiung].

In the past year, stool specimens of 1,732 alien laborers were examined for intestinal parasites. There was no significant difference in the positive rate between males and females. All alien laborers were examined in 3 hospitals and the positive rates were 24.8% (191/769), 13.6% (109/804) and 12.6% (20/159). Broken down by nationality, the positive rates in laborers from Thailand, Indonesia, the Philippines and Malaysia were 23.9% (192/803), 20.3% (62/305), 12.1% (60/498) and 4.8% (6/126) respectively. Of all alien laborers, 17.2% (297/1,732) had single infections of parasites and 1.3% (23/1,732) had multiple infections. Opisthorchis viverrinii was the most prevalent parasite found in infected alien laborers. 91.5% of alien laborers were aged from 21 to 40-years-old and had positive rates of parasites reaching 91.0%. The time of arrival in Taiwan had an effect in all alien laborers examined for parasites. The positive rate in laborers arriving during April to June was 22.7% which was significantly higher than the 16.3% for laborers arriving in other months.

Antibodies in serum of patients with clonorchiasis before and after treatment.

Sera of 31 patients infected with Clonorchis sinensis were examined using fraction 1 antigen by ELISA during a one-year observation. The results of ELISA with Igs, IgG and IgA demonstrated high sensitivity (100%, 100% and 90%) and specificity (100%, 100% and 87%). Sera specific Igs and IgG were significantly decreased in the 3rd month after treatment with praziquantel (25mg/kg body weight in one dose), and IgA was significantly decreased in the 1st month (paired t-test, p < 0.05). No eggs were found in the stool after treatment. Detection of sera specific Igs, IgG and IgA by ELISA was combined with stool examination to evaluate the effect of praziquantel and the completeness of the cure.