Antibodies against native proteins of Mycobacterium tuberculosis can detect pulmonary tuberculosis patients.

Accurate point-of-care testing (POCT) is critical for managing tuberculosis (TB). However, current antibody-based diagnosis shows low specificity and sensitivity. To find proper antigen candidates for TB diagnosis by antibodies, we assessed IgGs responsiveness to Mycobacterium tuberculosis proteins in pulmonary TB (PTB) patients. We employed major secreted proteins, such as Rv1860, Ag85C, PstS1, Rv2878c, Ag85B, and Rv1926c that were directly purified from M. tuberculosis. In the first screening, we found that IgG levels were significantly elevated in PTB patients only against Rv1860, PstS1, and Ag85B among tested antigens. However, recombinant PstS1 and Ag85B from Escherichia coli (E. coli) couldn't distinguish PTB patients and healthy controls (HC). Recombinant Rv1860 was not checked due to its little expression. Then, the 59 confirmed PTB patients from Soetomo General Academic Hospital, Surabaya, Indonesia, and 102 HC were tested to Rv1860 and Ag85B only due to the low yield of the PstS1 from M. tuberculosis. The ROC analysis using native Ag85B and Rv1860 showed an acceptable area under curve for diagnosis, which is 0.812 (95% CI 0.734-0.890, p < 0.0001) and 0.821 (95% CI 0.752-0.890, p < 0.0001). This study indicates that taking consideration of native protein structure is key in developing TB's POCT by antibody-based diagnosis.

Genetic diversity and relationship among Indonesian local sheep breeds on Java Island based on mitochondrial cytochrome b gene sequences.

BACKGROUND: The cytochrome b (Cyt b) gene is one of the most studied mitochondrial DNA (mtDNA) genes to determine sheep's genetic profile. This study aimed to determine the genetic diversity and relationships of several Indonesian local sheep populations on Java Island, Indonesia, based on the mtDNA Cyt b gene sequences. Blood samples were collected from forty-one individual sheep in seven populations of Indonesia local sheep breeds on Java Island (Priangan = 6, Garut = 6, Batur = 7, Wonosobo = 5, Javanese Thin-Tailed/JTT = 7, Javanese Fat-Tailed/JFT = 5, and Sapudi = 5). DNA extraction was performed on blood samples, and the mtDNA Cyt b gene was amplified using specific primers (Alek-CBF: 5'-CAACCCCACCACTTACAA-3' and Alek-CBR: 5'-CCTTGAGTCTTAGGGAGGTT-3'). The polymerase chain reaction (PCR) products were then sequenced, and data were analyzed using the MEGA version 7.0, DNA SP version 6.0, and NTSYS-pc version 2.11 software. RESULTS: A total of 1140 bp complete mtDNA Cyt b gene sequences in this study obtained 1134 monomorphic sites (I), six polymorphic sites (V), one segregation site (S), and five parsimony informative sites (P) with a nucleotide diversity (Pi), the average number of nucleotide differences (K), and sequence conservation (SC) were 0.00119, 1.35610, and 0.9947, respectively. There were six haplotypes consisting of two unique haplotypes and four shared haplotypes with a haplotype diversity (Hd) of 0.5415. The genetic distance within and between populations ranged from 0.0000 to 0.0016 and 0.0000 to 0.0020, respectively. Wonosobo, JFT, and Sapudi sheep have the closest relationship, and then these three breeds were close to JTT sheep, followed by Batur, Priangan, and Garut sheep. Two haplogroups have been found based on the Ovine haplogroup clustering. All Wonosobo, JTT, JFT, Sapudi sheep, and most Batur sheep were clustered into haplogroup B. In contrast, Garut sheep were mostly clustered into haplogroup A, while Priangan sheep were clustered into both haplogroups with the same percentage. CONCLUSION: Seven Indonesian local sheep breeds on Java Island have a close relationship clustered into two haplogroups, namely haplogroups A and B. Most Indonesian local sheep breeds on Java Island in this study were clustered into haplogroup B, except for Garut and Priangan sheep.

Genetic variability of the prion protein gene in Indonesian goat breeds.

Scrapie is a naturally occurring transmissible spongiform encephalopathy in sheep and goats. Resistance or susceptibility of small ruminants to classical scrapie is influenced by polymorphisms in the prion protein gene (PRNP). PRNP variability in Indonesian indigenous goat breeds has not been investigated so far and therefore was the goal of this study. Sanger sequencing of the PRNP gene coding region in 72 goats of the seven Indonesian breeds Kacang, Gembrong, Samosir, Kejobong, Benggala, Jawarandu, and Peranakan Etawah revealed three amino acid substitutions, namely W102G, H143R, and S240P. Some silent mutations were also found at codons 42 (a/g), 138 (c/t), and 179 (g/t). The PRNP alleles K222 and D/S146 known to have significant protective effects on resistance to classical scrapie in goats were not detected. The allele R143, which may have a moderate protective effect, had a frequency of 12% among the analyzed Indonesian goat breeds. While R143 was missing in Kacang and Benggala, its frequency was highest in the breed Gembrong (32%). No scrapie cases have been reported in Indonesia until now. However, in the case that selection for protective PRNP variants would become a breeding goal, the analyzed breeds will not be very useful resources. Other goat breeds which are present in the country should be investigated regarding resistance to scrapie, too.

Microbiome in sputum as a potential biomarker of chronicity in pulmonary resistant to rifampicin-tuberculosis and multidrug-resistant-tuberculosis patients.

Background: Cases of tuberculosis (TB) and multidrug-resistant TB (MDR-TB) in South-east Asia including Indonesia are still high. The presence of mixed infections in TB cases has been reported. Several studies revealed the role of the human microbiome in TB. This study purposes to characterize microbiome which can be a potential biomarker of chronicity in TB or MDR-TB. Methods: Sputum samples of pulmonary TB patients confirmed MDR-TB and resistant to rifampicin TB (RR-TB) were conducted Metagenomic next-generation sequencing. Principal coordinate analysis of UniFrac's showing the community structure of microbiome in MDR-TB comorbid diabetes mellitus (DM) is different from RR-TB noncomorbid DM (P = 0.003). Results: Proteobacteria microbiome in MDR-TB comorbid DM was more abundant than in RR-TB noncomorbid DM. Actinobacteria found in the small quantity in RR-TB and MDR-TB. Diversity of microbiome genera was greater in RR-TB. The linear discriminant analysis effect size analysis represents a genus biomarker whose abundance shows significant differences between groups, genus Rothia as a potential biomarker for RR-TB noncomorbid DM. Conclusions: Interesting findings is the community structure of microbiome in MDR-TB and RR-TB. In chronic TB such as recurrent, associated MDR-TB should attention to the findings of a small number of Actinobacteria could be a biomarker of TB which is also a determinant in patient taking combined anti-TB drugs of choice.

A POTENTIAL ZOONOTIC PARASITE: CRYPTOSPORIDIUM PARVUM TRANSMISSION IN RATS, PIGS AND HUMANS IN WEST LOMBOK, INDONESIA.

BACKGROUND: Cryptosporidium is a neglected zoonotic disease, but with the expansion of the human community into the animal environment, its incidence is increasing. Animals such as rats and pigs can act as intermediate hosts and transmit Cryptosporidium to humans due to their proximity. Transmission occurs due to the ability of Cryptosporidium to survive in any new host. The research aimed to identify and describe the transmission of Cryptosporidium from animals to humans. MATERIALS AND METHODS: This research was a cross sectional study and samples were collected from 84 rats caught in residential areas, 205 pigs, and 438 humans in West Lombok. Fecal samples were examined using polymerase chain reaction (PCR) and sequencing to isolate the presence of Cryptosporidium, and identify the genetic similarity of the parasites found in rats and pigs with those that infect humans. RESULTS: The PCR results found Cryptosporidium parvum in 4.76% (4/84) in rats; 6.34% 13/205) in pigs; and 0.91% (4/438) in humans. The sequencing results showed genetic kinship of C. parvum in rats, pigs, and humans. Based on sequence confirmation from Gene Banks and edited using ClustalW with MEGA X software, there are genetic similarities between Cryptosporidium isolates from West Lombok and C. suis isolates of cattle from Uganda and C. suis isolates of pigs from Slovakia. CONCLUSION: There are genetic similarities of Cryptosporidium in animals and humans, requiring that the Public Health programs in those contaminated areas must receive priority attention to prevent further transmission of these potentially fatal parasites.

Knowledge, Attitudes, and Behaviors on Utilizing Mobile Health Technology for TB in Indonesia: A Qualitative Pilot Study.

Tuberculosis (TB) infections remain a global health burden with a high incidence rate in South-East Asia, including Indonesia. TB control strategy is founded on early case detection and complete treatment to minimize transmission and prevent the emergence of drug resistance. However, many patients face challenges to comply with daily medication, causing many to adhere inconsistently or stop prematurely. Technological solutions could enhance adherence to treatment and support national screening and follow-up policies. These include telephone video communication, enabling health professionals to watch patients take their medication, address patients' concerns, and provide advice and support. This manuscript describes the outcome of a qualitative pilot study, based on a series of focus group discussions to assess the knowledge, attitudes, and behaviors, on the potential utilization of mobile technology for health purposes with a particular focus on TB treatment follow-up. The findings illustrate that general knowledge of mobile health technologies, of their legal framework of operations, and of their exact potential within the healthcare system is incomplete or poor. The novel findings are as follows: (a) the willingness of participants to learn about these technologies, (b) the open and welcoming attitude toward receiving such information even within frontline community settings, and (c) the willingness to back a government-supported, healthcare-driven set of such initiatives. Potential implementation barriers have also been highlighted. This study is an important first step toward understanding the attitudes and behaviors on utilizing mobile health technology for TB in Indonesia.

Gene expression tryptophan aspartate coat protein in determining latent tuberculosis infection using immunocytochemistry and real time polimerase chain reaction.

Background: Tuberculosis (TB) remains a major cause of morbidity and mortality worldwide. Problem of Latent Tuberculosis Infection (LTBI) is increasing in number especially in countries with high TB incidence rate, such as Indonesia. Although not every LTBI will become active TB, if untreated and not handled appropriately it can still be a source of transmission and may increase the rate of resistance to the first-line TB drugs. Mycobacterium tuberculosis as a cause of tuberculosis disease is an intracellular pathogens that survives within the phagosome of host macrophages. Several host factors are involved in this process, including the Tryptophan Aspartate-containing Coat Protein (TACO). TACO is a protein recruited and retained by viable Mycobacterium tuberculosis on the surface of the phagosome membrane to maintain its survival in phagosome, because the presence of TACO plays an important role in inhibiting the fusion of phagosomes and lysosomes. Objective: the aim of this studyis to assess the difference of gene expression TACO protein in Latent Tuberculosis Infection (LTBI) and healthy people. Method: A preliminary studyof mRNA examination of TACO protein using Immunocytochemistry (ICC) and Real Time-Polimerase Chain Reaction (RT-PCR) method by a PCR Light Cycler 2.0 machine (Roche) in LTBI and healthy groups. Results: 18 samples of peripheral blood monocyte cells (PBMCs) were collected and divided into 2 groups. We found that there was a significantly difference between the 2 groups of samples. Conclusion: Further research is required to consider that the measurement of TACO expression using RT-PCRcan used as one of the other method to determine LTBI.

The genetic profiles and maternal origin of local sheep breeds on Java Island (Indonesia) based on complete mitochondrial DNA D-loop sequences.

BACKGROUND AND AIM: Java Island is one of the islands in Indonesia which has local sheep breeds with specific characteristics and native development geography in certain regions. This study aimed to determine the genetic profiles and maternal origin of six local sheep breeds on Java Island. MATERIALS AND METHODS: This study was conducted by identifying the profiles of complete mitochondrial DNA (mtDNA) displacement loop (D-loop) region sequences on a total of 22 individual in six local sheep breeds on Java Island, including Javanese thin-tailed (JTT), Javanese Fat-Tailed (JFT), Batur (BTR), Wonosobo (WSB), Garut (GRT), and Priangan (PRG) sheep. The D-loop region was amplified using specific primers, and the polymerase chain reaction (PCR) was performed. The PCR products were purified and sequenced. RESULTS: The mtDNA D-loop analysis identified 21 haplotypes in the analyzed 22 animals with 123 polymorphic sites (V) consisting of 60 singleton variable sites (S) and 63 parsimony informative sites (P). Within all breeds tested, the haplotype diversity, the average number of pairwise differences (K), and nucleotide diversity (Pi) were 0.99567, 25.36364, and 0.02153, respectively. The genetic distance (D) within groups and between groups was 0.001-0.006 and 0.004-0.036, respectively. The phylogeny resulted in the presence of two haplogroups (Hap), which are 5 Hap A and 16 Hap B. All JTT, JFT, BTR, and WSB breeds were in the same cluster in Hap B, whereas GRT and PRG breeds were in clusters in both Hap A and Hap B. CONCLUSION: The high genetic diversity in six local sheep breeds on Java Island suggests that they originated from different genetic sources. JTT sheep have closer genetic relationships to JFT, BTR, and WSB sheep, and they are close to European sheep, whereas GRT sheep have closer genetic relationships to PRG sheep. Both are closer to Asian sheep than to European sheep.

The impact of religious festival on roadside livestock traders in urban and peri-urban areas of Yogyakarta, Indonesia.

BACKGROUND AND AIM: Eid al-Adha is one of the most important festivals celebrated by Muslims in Indonesia. Roadside livestock traders open their stalls during the Eid al-Adha period. This study aimed to investigate the characteristics and behaviors of roadside livestock traders in urban and peri-urban areas in Yogyakarta, Indonesia. MATERIALS AND METHODS: In-depth interviews with 36 roadside livestock traders were conducted on August 7-23, 2018 in urban (n=20) and peri-urban (n=16) areas of Yogyakarta. The collected data were analyzed by descriptive and statistical analysis using one-way analysis of variance. RESULTS: The results indicate that the trading activities of roadside livestock traders in urban areas last longer (p<0.05) than in peri-urban areas. No difference was found in the opening day of stalls, the number of buyers, and trends in animal prices set by roadside livestock traders in urban and peri-urban areas. Most traders sell sheep and goats, buy livestock at the animal market, and only open their stalls during Eid al-Adha. Prices are high in this period, and buyers directly visit the stalls. A significant difference exists in the selling price of livestock between Eid al-Adha and ordinary days (non-festival), and most roadside traders benefit from the Eid al-Adha momentum. CONCLUSION: Significant similarities exist among roadside livestock traders during the Eid al-Adha period in urban and peri-urban areas of Yogyakarta, Indonesia. Sheep are more desirable than goats and cattle in this period, and Eid al-Adha generates a high profit for roadside livestock traders.

EXPRESSION OF IMMUNOGLOBULIN, GRANZYME-B AND PERFORIN AGAINST Ag85A AND Ag85B PROTEINS OF MYCOBACTERIUM TUBERCULOSIS IN BALB/C MICE.

BACKGROUND: Ag85 is a protein that may maintain survival of M. tuberculosis in intracellular parts of host cells and is considered as a virulence factor. The expression of Ag85 protein can stimulate proliferation and differentiation of B- cells and T-cells in patients with tuberculosis. This research aimed to determine the ability of Ag85A and Ag85B proteins in activating the response of antibodies, granzyme-B and perforin in Balb/c mice. MATERIALS AND METHODS: Twenty-five male Balb/c mice were assigned into five groups. Group I was treated with adjuvant, group II with Bacillus Calmette-Guerin (BCG) vaccine, group III with a combination of BCG and Ag85A, group IV with a combination of BCG and Ag85B and group V with a combination of BCG, Ag85A and Ag85B. Concentrations of immunoglobulin G, granzyme-B and perforin were examined using ELISA and the number of CD8+ T-cells and NK T-cells were checked by flow cytometry. RESULTS: The highest concentration of immunoglobulin G was found in group V with 62.49±5.4327 ng/ml. The highest mean number of CD8+ T-cells, NK T-cells, granzyme-B and perforin was found in group IV with 4.32%, 1.03%, 35.11±1.7789 pg/ml and 6.19±0.2235 pg/ml, respectively. The results of One-Way ANOVA test showed that there were significant differences in immunoglobulin responses, with p<0.05. The expressions of granzyme-B and perforin were higher in mice treated with combination of BCG and recombinant proteins. CONCLUSIONS: Ag85 protein can be combined with the BCG vaccine to improve protection against M. tuberculosis infection.

Genetic variation and phylogenetic analysis of Indonesian indigenous catfish based on mitochondrial cytochrome oxidase subunit III gene.

AIM: This study aimed to analyze the genetic variation and phylogenetic reconstruction of Indonesian indigenous catfish using mitochondrial cytochrome oxidase subunit III sequences. MATERIALS AND METHODS: A total of 19 samples of catfish were collected from seven rivers (Elo [EM], Progo [PM], Kampar [KR], Musi [MP], Mahakam [MS], Kapuas [KS], and Bengawan Solo [BSBJ]) in five different geographical locations in Indonesia. The genome was isolated from the tissue. Mitochondrial DNA cytochrome oxidase subunit III was amplified using polymerase chain reaction (PCR) with CO3F and CO3R primers. The PCR products were sequenced and continued to analyze genetic variation and phylogenetic relationship using MEGA version 7.0 software. RESULTS: Cytochrome c oxidase (COX)-III gene sequencing obtained 784 nucleotides encoding 261 amino acids. Sequenced COX-III gene fragments were aligned along with other catfish from Genbank using ClustalW program and genetic diversity among species was analyzed using the MEGA Version 7.0 software. Among all samples, there were substitution mutations at 78 nucleotide sites, and there were 14 variations in amino acids. Catfish from PM, KR, MP, and KS had the same amino acids as Hemibagrus nemurus (KJ573466.1), while EM catfish had eight different amino acids and catfishBSBJhad 12 different amino acids. CONCLUSION: Indonesian catfish divided into four clades. BBSJ Catfish were grouped with Pangasianodon gigas, EM catfish were grouped with Mystus rhegma, and KS catfish were grouped with Hemibagrus spilopterus, while catfish MS, KR, PM, andMP were grouped with H. nemurus.

Characteristic profile of antibody responses to PPD, ESAT-6, and CFP-10 of Mycobacterium tuberculosis in pulmonary tuberculosis suspected cases in Surabaya, Indonesia.

Accurate and rapid diagnostic tools are important aspects of managing tuberculosis (TB) cases appropriately. However, the sensitivity and specificity of diagnostic kits based on immune response such as the tuberculin skin test (TST) and interferon gamma release assay (IGRA) are still debated. Thus, the exploration and assessment of specific biomarker-targeted antibodies are needed for the development of an accurate and rapid diagnostic tool. The present study was conducted in patients with a respiratory problem suspected to be TB at Dr. Soetomo Hospital, Surabaya, Indonesia. Among 102 patients tested by GeneXpert and AFB, 59 serum samples were from cases retrospectively determined to have active TB. A total of 102 serum of healthy controls (HC) was also collected. The PPD antigen and the recombinant CFP-10 and ESAT-6 proteins were prepared. Antibody responses against these proteins were evaluated by ELISA. All samples were also screened for the possibility of Mycobacterium avium-intracellulare complex (MAC) infection using Capilla MaC kit. The results showed that TB patients had a significantly higher concentration of IgG antibody in response to PPD than the HC. In addition, the receiver operating characteristic (ROC) curve analysis showed that PPD was acceptable for diagnostic purposes with an AUC value of 0.835 (95% CI 0.770-0.900, p < 0.0001). However, ESAT-6 and CFP-10 had low AUCs, and 32 samples from both groups showed a low concentration of IgA antibody against all antigens. The MAC detection results also showed that the concentration of IgA in the HC group was the highest. The current results indicate that PPD is a better antigen for antibody-based detection of TB than ESAT-6 and CFP-10. Based on the MAC detection assay, 53 people in the HC group were probably infected with rapidly growing nontuberculous mycobacteria (NTM), although antibody response to PPD was low.

Characteristic profile of antibody responses to PPD, ESAT-6, and CFP-10 of Mycobacterium tuberculosis in pulmonary tuberculosis suspected cases in Surabaya, Indonesia

Abstract Accurate and rapid diagnostic tools are important aspects of managing tuberculosis (TB) cases appropriately. However, the sensitivity and specificity of diagnostic kits based on immune response such as the tuberculin skin test (TST) and interferon gamma release assay (IGRA) are still debated. Thus, the exploration and assessment of specific biomarker-targeted antibodies are needed for the development of an accurate and rapid diagnostic tool. The present study was conducted in patients with a respiratory problem suspected to be TB at Dr. Soetomo Hospital, Surabaya, Indonesia. Among 102 patients tested by GeneXpert and AFB, 59 serum samples were from cases retrospectively determined to have active TB. A total of 102 serum of healthy controls (HC) was also collected. The PPD antigen and the recombinant CFP-10 and ESAT-6 proteins were prepared. Antibody responses against these proteins were evaluated by ELISA. All samples were also screened for the possibility of Mycobacterium avium-intracellulare complex (MAC) infection using Capilla MaC kit. The results showed that TB patients had a significantly higher concentration of IgG antibody in response to PPD than the HC. In addition, the receiver operating characteristic (ROC) curve analysis showed that PPD was acceptable for diagnostic purposes with an AUC value of 0.835 (95% CI 0.770-0.900, p < 0.0001). However, ESAT-6 and CFP-10 had low AUCs, and 32 samples from both groups showed a low concentration of IgA antibody against all antigens. The MAC detection results also showed that the concentration of IgA in the HC group was the highest. The current results indicate that PPD is a better antigen for antibody-based detection of TB than ESAT-6 and CFP-10. Based on the MAC detection assay, 53 people in the HC group were probably infected with rapidly growing nontuberculous mycobacteria (NTM), although antibody response to PPD was low.

SEROPREVALENCE AND RISK FACTOR OF TOXOPLASMOSIS IN SCHIZOPHRENIA PATIENTS REFERRED TO GRHASIA PSYCHIATRIC HOSPITAL, YOGYAKARTA, INDONESIA.

BACKGROUND: Toxoplasmosis is an infectious disease caused by protozoan parasite called Toxoplasma gondii. Toxoplasma gondii is an intracellular protozoan parasite belong to phylum Apicomplexa, is an obligate parasite in mammals. The active proliferating trophozoites or tachyzoites are usually seen in the acute stage of infection, while the resting bradyzoites formed tissue cysts are primary found in muscle and brain. Human infection occurs mainly by ingesting food or water contaminated with oocyst or eating an undercook meat containing tissue cyst. Human might be infected via blood transfusion, organ transplantation or transplacenta transmission. Schizophrenia is a complex neuropsychiatric disease of the central nervous system, which contributing to behavioral changes which may resulted in higher risk to T. gondii infection. The purpose of this study were to know difference of seroprevalence and risk factor of toxoplasmosis between schizophrenia group and control group. MATERIALS AND METHODS: Serum sample were collected 94 among schizophrenia patient at Grhasia Hospital and 64 normal population (control group). Antibody IgG of T. gondii was measured using ELISA method (Enzym Link Immnusorbent Assay) and questionnaires were used to collect risk factor data among the respondent. RESULTS: The seroprevalence antibody IgG of patient with schizophrenia (69.14%) higher than control group (65.625%), but not significantly different (p>0.05). There was an association between some of risk factor with seropositive of toxoplasmosis in both group. In schizophrenia group, risk factor that associated with toxoplasmosis are uncooked meat consumption, contact with uncooked meat and soil, handwashing habit, uncooked water consumption, and water source. In control group, risk factor that associated are having cattles/pet, undercook meat consumption, and water source. CONCLUSION: This finding have shown seroprevalence of schizophrenia group higher than non-schizophrenia group and risk factor which associated with toxoplasmosis was different between two groups.

Regulatory elements of stx2 gene and the expression level of Shiga-like toxin 2 in Escherichia coli O157:H7.

BACKGROUND/PURPOSE: Shiga-like toxin (Stx) is an important factor in the pathogenesis of Escherichia coli O157:H7 infection and is responsible for some severe complications. Stx2 is usually associated with hemolytic uremic syndrome in humans. Its expression is regulated by elements located upstream of the stx2 gene, including stx2-promoter sequence, ribosome binding site, and the antiterminator q gene. The present study aimed to find the correlation between regulatory elements and the expression level of Stx2 in two local isolates of E. coli O157:H7. METHODS: Two local E. coli O157:H7 strains SM-25(1) and KL-48(2), originating from human and cattle feces, respectively, and an E. coli reference strain, ATCC 43894, were investigated. The complete stx2 gene covering the sequences of promoter, ribosome binding site, and open reading frame and q gene of each strain was analyzed. The magnitude of Stx2 production was detected with a reverse passive latex agglutination method and Stx mediated cellular damage was determined with the Vero cell assay. RESULTS: A comparison of the complete stx2 gene contained stx2-promoter, ribosome binding site, and q genes of two local strains KL-48(2) and SM25(1), and the E. coli ATCC 43894 showed that the amino acid sequences were identical. Both local isolates were Stx negative in the reverse passive latex agglutination test and nontoxic in the Vero cell assay. CONCLUSION: The expression level of Shiga-like toxin of the two local isolates of E. coli O157:H7 did not only depend on the regulatory elements of the stx2 gene.

Prevalence and Risk Factors for Toxoplasmosis in Middle Java, Indonesia.

Toxoplasmosis is a zoonosis caused by Toxoplasma gondii. Risk factors include consumption of undercooked meat, raw vegetables, and unfiltered water. This study aims to determine the seroprevalence and spatial distribution of toxoplasmosis in Middle Java, Indonesia, using an EcoHealth approach, combined with geographic information system (GIS). A total of 630 participants were randomly selected from seven districts. Each participant completed a questionnaire and provided a blood sample. The seroprevalence of toxoplasmosis was 62.5%. Of those who were seropositive, 90.1% were IgG+, and 9.9% were IgG+ and IgM+. Several risk factors were identified, including living at elevations of ≤200 m, compared with >200 m (OR = 56.2; P < 0.001), daily contact with raw meat (OR = 1.8; P = 0.001), unfiltered water (OR = 1.7; P = 0.003), and density of cats (OR = 1.4; P = 0.045). Visualizing the spatial distribution of seropositive respondents highlighted clustering in lowland areas. This study highlighted that Middle Java has a high prevalence of toxoplasmosis and identified some important environmental, ecological, and demographic risk factors. When researching diseases, such as toxoplasmosis, where animal hosts, human lifestyle, and environmental factors are involved in transmission, an EcoHealth method is essential to ensure a fully collaborative approach to developing interventions to reduce the risk of transmission in high-risk populations.