The efficacy of agar-plate culture has been evaluated for the detection of Strongyloides stercoralis and hookworm, compared with direct smear, the formalin-ether sedimentation technique and the filter-paper method. Of 1085 stool samples from the routine laboratory service at King Chulalongkorn Memorial Hospital in Bangkok, 241 samples harboured S. stercoralis, 153 hookworm and 2 Rhabditis hominis. The recovery rate of S. stercoralis by agar-plate culture is significantly superior to the other methods (P < 0.005). The ratios of positive results from the methods used to the total number of S. stercoralis-positive cases were as follows: 1:1.03 by agar-plate culture, 1:1.85 by the filter-paper method, 1:1.98 by the sedimentation technique and 1:10.48 by direct stool smear. A similar trend of the efficacy ratio of each method was obtained for hookworm detection. The characteristic furrows left by hookworm larvae, and larvae and adults of S. stercoralis could be used for preliminary species identification. Daily search for furrows on agar plates for up to 6 consecutive days resulted in an increased sensitivity for diagnosis of both S. stercoralis and hookworm infections.

To investigate the extent of genetic heterogeneity in the genus Isospora infecting patients in Thailand, a total of 38 fecal samples containing Isospora oocysts from human immunodeficiency virus/acquired immunodeficiency syndrome patients (n = 30), corticosteroid-treated patients (n = 3) and immunocompetent individuals (n = 5) were recruited for analysis. Remarkable variation in the maximum width and length of oocysts both within and between isolates was observed. However, the average length-width ratio of oocysts was within the range for I. belli (> 1.2). Ex vivo sporogonic development of freshly passed oocysts in feces from three of these isolates was observed longitudinally, showing that 27% of these oocysts underwent complete sporulation. Interestingly, 95% of sporulated oocysts contained two sporocysts in an oocyst with four sporozoites in each sporocyst, and Caryospora-like oocysts, characterized by eight sporozoites enclosed by a single sporocyst, were also detected (5%). The small subunit ribosomal RNA (rRNA), 5.8S rRNA, internal transcribed spacer 1 (ITS-1), and ITS-2 were highly conserved, indicating that there were no cryptic species or extensive strain variation.

SummaryWe isolated Acanthamoebae from the first two keratitis patients identified in Thailand in 1988 and 1990. The patients developed decreased vision, severe photophobia, severe eye pain and foreign body sensation after minor corneal trauma. The lesions included generalized superficial punctate keratitis, stromal corneal ulcer with keratic precipitate and uveitis in one case, and corneal ulcer with abscess in the other. Both cases were diagnosed by isolation of characteristic trophozoites and cysts of Acanthamoeba from corneal tissue by nonnutrient agar culture method. Based on cyst morphology, A. castellanii and A. polyphaga were detected in one case, and A. castellanii and A. triangularis in the other. Restriction fragment length polymorphism analysis of mitochondrial DNA (mtDNA-RFLP) revealed that each patient harboured a single parasite population. One shared mtDNA-RFLP with an authentic strain of A. castellanii, and the other gave a new unique pattern. Thus species identification of Acanthamoeba based on cyst morphology per se can be arbitrary, and mtDNA-RFLP may be more appropriate for accurate species/strain differentiation amongst morphologically heterogeneous populations of Acanthamoebae.