Intestinal Helminths Recovered from Humans in Xieng Khouang Province, Lao PDR with a Particular Note on Haplorchis pumilio Infection.

A survey of intestinal helminths was undertaken in riparian people in Xieng Khouang Province, Lao PDR. Fecal specimens were collected from 643 people (289 males and 354 females) residing in 4 districts (Nonghet, Kham, Phoukout, and Pek) and were examined by the Kato-Katz technique. The overall helminth egg positive rate was 41.2%, and hookworms revealed the highest prevalence (32.7%) followed by Trichuris trichiura (7.3%) and Ascaris lumbricoides (5.6%). The positive rate for small trematode eggs (STE), which may include Opisthorchis viverrini, heterophyids, and lecithodendriids, was 4.4%. For recovery of adult helminths, 12 STE or nematode/cestode egg-positive people were treated with 40 mg/kg praziquantel and 15 mg/kg pyrantel pamoate, and then purged. Mixed infections with 2 Haplorchis species (H. pumilio and H. taichui), Centrocestus formosanus, Opisthorchis viverrini, a species of cestode (Taenia saginata), and several species of nematodes including hookworms and Enterobius vermicularis were detected. The worm load for trematodes was the highest for H. pumilio with an average of 283.5 specimens per infected person followed by C. formosanus, H. taichui, and O. viverrini. The worm load for nematodes was the highest for hookworms (21.5/infected case) followed by E. vermicularis (3.2/infected case). The results revealed that the surveyed areas of Xieng Khouang Province, Lao PDR are endemic areas of various species of intestinal helminths. The STE found in the surveyed population were verified to be those of heterophyids, particularly H. pumilio.

Fishborne trematode metacercariae in Luang Prabang, Khammouane, and Saravane Province, Lao PDR.

Fishborne trematode (FBT) metacercariae were investigated in fish from 3 Provinces of Lao PDR. Total 242 freshwater fish of 40 species were collected in local markets of Luang Prabang (59 fish of 16 species), Khammouane (81 fish of 19 species), and Saravane (97 fish of 14 species), and each of them was examined by artificial digestion method. Four species of metacercariae (Opisthorchis viverrini, Haplorchis taichui, Haplorchis yokogawai, and Centrocestus formosanus) were detected. O. viverrini was detected in 35 fish (14.5%), and their density was 252 per infected fish (Luang Prabang, 88 metacercariae in 5 fish; Khammouane, 187 in 6 fish; Saravane, 303 in 24 fish). H. taichui was found in 102 fish (42.1%), and their density was 485 per infected fish (Luang Prabang, 260 metacercariae in 38 fish; Khammouane, 1,084 in 23 fish; Saravane, 359 in 41 fish). H. yokogawai was detected in 92 fish (38.0%), and their density was 222 per infected fish (Luang Prabang, 362 metacercariae in 17 fish; Khammouane, 126 in 20 fish; Saravane, 214 in 55 fish). Metacercariae of C. formosanus were found in 8 fish (3.3%), and their density was 3 per infected fish. In the present study, it has been confirmed that FBT metacercariae, in particular, H. taichui, H. yokogawai, and O. viverrini, are highly prevalent in fish from Luang Prabang, Khammouane, and Saravane Province, Lao PDR.

Prevalence of the intestinal flukes Haplorchis taichui and H. yokogawai in a mountainous area of Phongsaly Province, Lao PDR.

Phongsaly Province, located in the northernmost area of Lao PDR, was previously suggested to be endemic for the liver fluke Opisthorchis viverrini infection. To confirm, or rule out, this suggestion, the Phonxay village in the Khoua District, Phongsaly Province, was selected for a survey. Ten volunteers (8 men and 2 women aged 31-57 years) who consumed raw freshwater fish and had gastrointestinal troubles were treated with a single dose of praziquantel (40 mg/kg) and pyrantel pamoate (10 mg/kg) and purged with magnesium sulfate to recover any worm parasites. Eight of the 10 volunteers expelled 1 or more species of trematodes, nematodes, or cestodes (worm positive rate; 80%). The worms were morphologically identified as H. taichui (861 worms from 8 people), H. yokogawai (59 from 6 people), Phaneropsolus bonnei (1 from 1 person), Trichostrongylus sp. (2 from 2 people), Ascaris lumbricoides (2 from 1 person), Enterobius vermicularis (11 from 3 people), and Taenia saginata (1 strobila with scolex from 1 person). The results indicate that the mountainous area of Phongsaly Province, Lao PDR, is not endemic for the liver fluke but endemic for intestinal flukes, in particular, Haplorchis taichui and H. yokogawai.

Ultrasonographic investigation of cholangiocarcinoma in Lao PDR.

Opisthorchis viverrini is a group 1 carcinogen that causes cholangiocarcinoma (CCA). Although opisthorchiasis is known to be severely endemic to several areas along the Mekong River in Lao PDR, the CCA status of residents of this region is still under investigation. In this study, we analyzed the results of abdominal ultrasonography (US) performed on 6113 residents in 9 provinces (Vientiane Municipality, Savannakhet, Phongsaly, Khammouane, Saravane, Champasak, Vientiane, Xieng Khuouang, and Luang Prabang provinces) of Lao PDR from 2007 to 2011. Overall, 51 cases (0.83%) were detected with suspected CCA. The CCA rates in Vientiane Municipality and in Savannakhet and Khammouane provinces were 1.45%, 1.58%, and 1.09%, respectively. However, in the other 6 provinces, the rate of CCA averaged only 0.26%. In the 3 provinces with higher rates of CCA, bile duct dilatation (grade ≥ 2) was also significantly more prevalent (P < 0.0001). These results are concordant with previous reports showing a higher endemicity of opisthorchiasis in Vientiane Municipality and in Savannakhet and Khammouane provinces.

Application of dipstick dye immunoassay (DDIA) kit for the diagnosis of schistosomiasis mekongi.

The dipstick dye immunoassay (DDIA), developed in China for the detection of antibodies against Schistosoma japonicum, relies on soluble egg antigen (SEA) labelled with a colloidal dye. This assay is not only rapid, simple and inexpensive, but also particularly useful for screening in the field. In order to determine whether S. japonicum antigens are sufficiently cross-reactive to make the assay applicable for the diagnosis also of S. mekongi a DDIA approach based on the S. japonicum SEA was tried in cohorts of healthy and infected people living in areas non-endemic and endemic with regard to schistosomiasis mekongi in Cambodia and Laos. A sensitivity of 97.1% was recorded when testing Cambodian subjects, correctly diagnosing 33 out of 34 infected people. When the assay was applied in Laos, a sensitivity of 98.6% (69/70) was found. None of 114 residents living in a non-endemic area in Cambodia tested positive. A cross-reaction of 18.3% was found in patients infected with Opisthorchis viverrini. The results support the notion that the DDIA using S. japonicum SEA antigens can safely be implemented for the diagnosis of schistosomiasis mekongi, but care is needed in the interpretation of results obtained from areas that are co-endemic for O. viverrini.

Pathogenicity of Shigella in healthy carriers: a study in Vientiane, Lao People's Democratic Republic.

Shigella spp. isolated from diarrheal patients and non-diarrheal carriers were examined by PCR for the presence of two pathogenic genes, chromosomal ipaH and invasive plasmid encoded ial. Shigella spp. were detected in 7 of 72 diarrheal cases examined (9.7%), and 9 of 145 non-diarrheal cases (6.2%). All isolates from diarrheal cases harbored both ipaH and ial, while all isolates from non-diarrheal cases were positive for ipaH but not ial. These results suggested that Shigella spp. in healthy carriers were basically non-pathogenic.

Study on cyclospora cayetanensis associated with diarrheal disease in Nepal and Loa PDR.

A study of diarrheal diseases associated with Cyclospora cayetanensis was conducted in Nepal and Lao PDR. A total of 2083 samples were included in this study. Samples in Nepal were collected from October 1999 to August 2002 whereas samples in Lao PDR were collected from February 2002 to June 2003. C. cayetanensis was detected by direct microscopy using ultraviolet and differential interference contrast microscopy. The overall positive rate in Nepal was 9.2% (128/ 1397). A higher positive rate was observed in children aged 10 years and under (11.1%) and was lowest in the age group of 51-60 years (3.1%). A significantly higher positive rate was observed in the summer (rainy season) (12.6%) with the lowest prevalence in the spring (dry season) (1.8%) (p < 0.05). The positive rate was closely associated with rainfall (ml/month). Interestingly, only one of the total 686 samples (0.1%) from Lao PDR was found to be positive for Cyclospora oocysts.

The importance of leptospirosis in Southeast Asia.

The importance of leptospirosis in Southeast Asia was assessed in conjunction with other studies supported by the U.S. Naval Medical Research Unit No. 2 (US NAMRU-2), Jakarta, Republic of Indonesia. These included studies of hospital-based, acute clinical jaundice in Indonesia, Lao PDR, and Socialist Republic of Vietnam; nonmalarial fever in Indonesia; and hemorrhagic fever in Cambodia. Background prevalence estimates of leptospiral infection were obtained by a cross-sectional, community-based study in Lao PDR. Laboratory testing methods involved serology, microscopic agglutination test, and reverse-transcriptase polymerase chain reaction. Suggestive evidence of recent leptospiral infections was detected in 17%, 13%, and 3% of patients selected on the basis of non-hepatitis A through E jaundice, nonmalarial fever, and hemorrhagic fever (in the absence of acute, dengue viral infections). Leptospiral IgG antibody, reflective of prior infections, was detected in 37% of human sera, collected in Lao PDR. The predominant leptospiral serogroups identified from cases with clinical jaundice were Hurstbridge, Bataviae, and Icterohaemorrhagiae tonkini LT 96 69. Among the nonmalarial febrile cases, Bataviae was the most frequently recognized serogroup. Pyrogenes and Hurstbridge were the principal serogroups among the hemorrhagic fever case subjects. These findings further attest to the relative importance of clinical leptospirosis in Southeast Asia. The wide spectrum of clinical signs and symptoms associated with probable, acute, leptospiral infections contributes to the potential of significant underreporting.

The incidence of Escherichia coli having pathogenic genes for diarrhea: a study in the People's Democratic Republic of Lao.

The incidence of Escherichia coli having pathogenic genes for diarrhea was studied in Laos in 2002. A total of 525 E. coli strains from 278 patients (basically, two E. coli isolates from each patient) were examined by PCR to detect the known pathogenic genes (stx, eae, elt, est, ipaH, and aggR). These genes were detected in 23 strains from 16 patients (16/278: 5.8%). In 10 cases of the 16, one of the two isolates from each individual was negative for the gene, and in the other six cases, both isolates had the gene (same gene in four cases). E. coli having eae but no stx (enteropathogenic E. coli [EPEC]) was found in two cases out of 278 (0.7%). Nevertheless, Class I classical EPEC (serogroup-based) was found in 77 cases (28%). Enterotoxigenic E. coli, enteroaggregative E. coli, and enterohemorrhagic E. coli were found in 9, 4, and 1 cases, respectively. Enteroinvasive E. coli was not detected. This study suggested that the incidence of diarrhea due to E. coli is not as high as has been previously thought.

Analysis of the effectiveness of control measures against Schistosoma mekongi using an intra- and inter-village model in Champasak Province, Lao PDR.

Schistosomiasis mekongi is prevalent in the Khong district of Lao PDR, made up of one big island, Khong, and numerous small islands in the Mekong River. Schistosoma mekongi is spread by Neotricula aperta as the intermediate host along the Mekong River. Therefore, even if an epidemic of S. mekongi were stamped out in a certain village, infection may recur if the source of infection is a village located in the upper reaches of the Mekong River. The purpose of this study was to construct a mathematical model for the transmission of S. mekongi among villages from the upper to lower Mekong River to estimate the effect of control measures against it. The chief characteristic of the present model is competence in dealing with the spread of infection among villages through the Mekong River in consideration of the reduction in longevity of cercariae and miracidia and their diffusion in the river. The model also takes into account seasonal fluctuation in the water level of the Mekong River, which affects human behavior in terms of water contact. The results of simulations indicated that the prevalence of schistosomiasis mekongi would be suppressed to a low level for a long time in a village further downstream when universal mass treatment is performed in villages further upstream simultaneously.

Antibiotic resistance conferred by a class I integron and SXT constin in Vibrio cholerae O1 strains isolated in Laos.

Changes in the drug susceptibility pattern were observed in Vibrio cholerae O1 isolated in the Lao People's Democratic Republic during 1993 to 2000. In this study, 50 V. cholerae O1 strains were selected during this period for studying the presence of class I integron and SXT constin. Twenty-four streptomycin-resistant strains out of 26 isolated before 1997 contained a class I integron harboring the aadA1 gene cassette. Twenty-four strains isolated after 1997 contained an SXT constin (a large conjugative element). Twenty of the strains were resistant to chloramphenicol, tetracycline, streptomycin, and trimethoprim-sulfamethoxazole, while four strains were susceptible to the antibiotic tested. The resistance genes included in the SXT constins were floR, tetA, strAB, and sulII, which encode resistance to chloramphenicol, tetracycline, streptomycin, and sulfamethoxazole, respectively. The antibiotic resistance gene cluster was found to be deleted in the four susceptible strains. SXT(LAOS) did not contain dfrA1 or dfr18, which confer resistance to trimethoprim in SXT(ET) and SXT(MO10), respectively. A hot spot region of SXT(LAOS) was sequenced, and we identified two novel open reading frames showing homology to sO24 (exonuclease) and sO23 (helicase) of the genomic island associated with the multidrug resistance region of Salmonella enterica serovar Typhimurium DT104. Analysis of SXT(LAOS) showed that there is a continuous flux of genes among V. cholerae SXT constins which should be carefully monitored.

Staphylococcus aureus isolated from hospital staff: a comparative study of Laos and Japan.

Staphylococcus aureus isolated from hospital staff (nurses and doctors) in Laos were characterized for future reference and for comparison with organisms found in Okinawa, Japan. No methicillin-resistant S. aureus (MRSA), among 38 isolates examined, was found in Laos, but 16 of 36 Japanese isolates (44%) were identified as MRSA. The minimum inhibitory concentrations (MICs) of ampicillin and tetracycline against Lao isolates were definitely higher than those against Japanese isolates. The MIC(80) of tetracycline was 50 microg/ml and 0.4 microg/ml in Lao and Japanese isolates, respectively. There was no marked difference in the production of enterotoxin by S. aureus strains in the two countries. Toxic shock syndrome toxin-1 was produced by only 1 of 38 Lao isolates but by 5 of 36 Japanese isolates. Coagulase types varied greatly, but about 50% of the isolates in both countries produced untypable coagulase. Since the interaction of Laotians with foreigners has increased rapidly in recent years, the careful monitoring of drug-resistant pathogens is required.