Seroepidemiological Survey of Human Immunodeficiency Virus (HIV), Hepatitis B Virus (HBV) and Hepatitis C Virus (HCV) Infections among The Hill Tribes in Northern Thailand

We report the results of seroepidemiological survey of Human immunodeficiency virus (HIV), Hepatitis B virus (HBV) and Hepatitis C virus (HCV) infections among the Karen, La-Wah and Lahu-Na, designated as the hill tribes, in northern Thailand. Some of these hill tribes are living in the remote and isolated mountain areas, settling their own communities. Anti-HIV seropositive cases were found only in the Lahu-Na (2.6%). The highest incidence of Hepatitis B surface antigen (HBs-Ag) positive was found in the Karen (13.2%), followed by the Lahu-Na (2.6%) and the La-Wah (1.7%). The highest incidence of anti-Hepatitis C virus antibody (anti-HCV) positive was found in the La-Wah (3.3%), followed by the Karen and the Lahu-Na (2.6%, respectively). Two out of nine anti-HCV positive cases were from seven and 11 year-old Karenean girls, who had no previous history of surgery, blood transfusion, intravenous medication, vaccination and dental therapy. These results suggest that HIV infections have not yet reached to the hill tribes, except the Lahu-Na. One of the possible transmission routes of HCV infection is a vertical or intrahhousehold infection among the hill tribes in northern Thailand

High prevalence of Haplorchis taichui metacercariae in cyprinoid fish from Chiang Mai Province, Thailand

This study aimed to investigate Haplorchis taichui metacercarial infection in fish collected from the Chom Thong and Mae Taeng districts, Chiang Mai Province during November 2001 to October 2002. A total 617 cyprinoid fish of 15 species were randomly collected and examined for H. taichui metacercariae. All the species of fish were found to be infected with H. taichui. The infection rates were 91.4% (266/290) and 83.8% (274/327), with mean intensities of 242.9 and 107.4 in the Chom Thong and Mae Taeng districts, respectively. The portion of the fish body with the highest metacercarial density was the muscles, and second, the head, in both districts. In addition, the fish had mixed-infection with other species of trematodes, namely: Centrocestus caninus, Haplorchoides sp, and Haplorchis pumilio

Susceptibility of conidia from <i>P. marneffei</i> wild type, <i>sakA</i> mutant (Δ<i>sakA</i>) and <i>atfA</i> mutant (Δ<i>atfA</i>) to UV light.

<p>Conidia of each strain were plated in duplicate on SDA and exposed to different UV light radiation at 0, 2000, 4000, 6000 and 8000 microjoules/cm². Data are from three independent experiments and standard error bars of the mean bars are shown (p<0.05).</p

High prevalence of Haplorchis taichui metacercariae in cyprinoid fish from Chiang Mai Province, Thailand

This study aimed to investigate Haplorchis taichui metacercarial infection in fish collected from the Chom Thong and Mae Taeng districts, Chiang Mai Province during November 2001 to October 2002. A total 617 cyprinoid fish of 15 species were randomly collected and examined for H. taichui metacercariae. All the species of fish were found to be infected with H. taichui. The infection rates were 91.4% (266/290) and 83.8% (274/327), with mean intensities of 242.9 and 107.4 in the Chom Thong and Mae Taeng districts, respectively. The portion of the fish body with the highest metacercarial density was the muscles, and second, the head, in both districts. In addition, the fish had mixed-infection with other species of trematodes, namely: Centrocestus caninus, Haplorchoides sp, and Haplorchis pumilio

Susceptibility to oxidative stresses of <i>P. marneffei</i>.

<p>Growth of <i>P. marneffei</i> wild type (F4), the <i>atfA</i> mutant (SC) and <i>atfA</i> complemented strain (AC1) at 25°C and 37°C on MM agar supplemented with 2 and 0.5 mM <i>t</i>-BOOH (B and F), 2 and 1 mM H₂O₂ (C and G), and 0.25 mM and 25 µM menadione (D and H). Five microliters of cell dilutions (5×10⁴ to 5 cells) were inoculated on MM agar containing each compound. (A) and (E) represent MM control plates at 25°C and 37°C, respectively.</p

<i>atfA</i> expression during phase transition.

<p>RNA was isolated from <i>P. marneffei</i> strain F4 cells including conidia collected from cultures grown for seven days on PDA at 25°C, three days in SDB at 25°C (mycelia), and six days in BHI broth at 37°C (yeast). 18S rRNA was used as loading control of each growth phase.</p

Deletion of <i>atfA</i> does not affect chitin deposition and cell wall integrity.

<p>(A) <i>P. marneffei</i> wild type (F4), <i>atfA</i> mutant (SC) and <i>atfA</i> complemented (AC1) strains were grown for seven day at 25°C on PDA and stained with CFW day four and day seven to visualize cell wall and septa. Scale bar represents five micrometers. (B to M) five microliters of cell dilutions (5×10⁴ to 5 cells) of wild type, <i>atfA</i> mutant and <i>atfA</i> complemented strains were inoculated on media supplemented with 5 µg/ml CFW(C and I), 0.004% SDS (D and J), 10 µg/ml and 4 µg/ml amphotericin B (F and L) and 8 µg/ml and 0.8 µg/ml itraconazole (G and M). (B and E) and (H and K) represent MM control plates at 25°C and 37°C, respectively.</p

<i>atfA</i> gene is not involved in osmotic and heat stress responses in <i>P. marneffei</i>.

<p>Five microliters of cell dilutions (5×10⁴ to 5 cells) of wild type (F4), <i>atfA</i> mutant (SC) and <i>atfA</i> complemented (AC1) strains were inoculated on MM agar supplemented with 0.5 and 0.25 M NaCl (B and E) and 1 M sorbitol (C and F). (G) MM agar was incubated at 39°C. (A) and (D) represent MM control plates at 25°C and 37°C, respectively.</p

Strategy for construction of the <i>atfA</i> complemented strain.

<p>(A) The restriction map demonstrates recognition sites of <i>Eco</i>RV (EV) and <i>Bss</i>SI (BsI) used in Southern blot analysis to detect the presence of <i>atfA</i> gene in the <i>atfA</i> complemented strain (AC1) comparing to the wild type strain (WT). The grey bar indicates the position of probe that is specific to the <i>atfA</i> gene. (B) Southern blot hybridization of genomic DNA from the wild type (F4) and the <i>atfA</i>-complemented (AC1) strains using probe in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0111200#pone-0111200-g002" target="_blank">Figure 2A</a>. Probe (a 0.9 kb fragment of <i>atfA</i>) hybridized with 5.6 kb fragment and unpredicted fragment (10.0 kb fragment) of <i>Eco</i>RV- and BssSI-digested DNA from the wild type strain, respectively. For the <i>atfA</i> complemented strain, probe hybridized with unpredicted fragment (3.5 kb fragment) and 7.5 kb fragment of <i>Eco</i>RV- and BssSI-digested DNA from the <i>atfA</i> complemented strain, respectively.</p

Gene expressions and survival of <i>P. marneffei</i> under heat stress at 42°C.

<p>(A) Relative RNA expression of <i>sakA</i> and <i>atfA</i> genes of conidia from <i>P. marneffei</i> wild type determined by real-time PCR. Conidia were incubated at 42°C for 10, 20, 30 or 40 minutes. Total RNA was extracted from conidia and subjected to real-time PCR. Expression level of heat stress cells is presented as relative value to the expression level from no stress cells which is given a value of 1. GAPDH gene expression level was used to normalize amounts of input RNA. (B) Survival of conidia from <i>P. marneffei</i> wild type (WT), <i>atfA</i> mutant (Δ<i>atfA</i>) and complemented strains (Δ<i>atfA</i> + <i>atfA</i>) after incubating in BHI at 42°C for one hour. (C) Relative RNA expression level of <i>atfA</i> gene of conidia from <i>P. marneffei</i> wild type (WT), <i>sakA</i> mutant (Δ<i>sakA</i>). Conidia were incubated at 42°C for 20 minutes and total RNA was extracted and subject to real-time PCR. Results were obtained from three independent experiments and standard error bars of the mean bars are shown (p<0.05).</p