Pythiosis is a life-threatening infectious disease of humans and animals caused by the oomycete microorganism Pythium insidiosum. The disease has been increasingly diagnosed worldwide. P. insidiosum inhabits freshwater and presents in two forms: mycelium and zoospore. Clinical manifestations of pythiosis include an infection of the artery, eye, skin, or gastrointestinal tract. The management of pythiosis is problematic due to the lack of effective treatment. Many patients die from an uncontrolled infection. The drug susceptibility testing provides clinicallyuseful information that could lead to proper drug selection against P. insidiosum. Currently, no standard CLSI protocol for the drug susceptibility of P. insidiosum is available. This review aims at describing methods and antimicrobial agents for susceptibility testing against P. insidiosum. Several in-house in vitro susceptibility methods (i.e., broth microdilution method, radial growth method, and agar diffusion method) have been established for P. insidiosum. Either mycelium or zoospore can be an inoculum. Rabbit is the commonly-used model of pythiosis for in vivo drug susceptibility testing. Based on the susceptibility results (i.e., minimal inhibitory concentration and inhibition zone), several antibacterial and antifungal drugs, alone or combination, exhibited an in vitro or in vivo effect against P. insidiosum. Some distinct compounds, antiseptic agents, essential oils, and plant extracts, also show anti-P. insidiosum activities. Successfully medical treatment, guided by the drug susceptibility data, has been reported in some pythiosis patients. Future studies should emphasize finding a novel and effective anti-P. insidiosum drug, standardizing in vitro susceptibility method and correlating drug susceptibility data and clinical outcome of pythiosis patients for a better interpretation of the susceptibility results.

Pythium insidiosum is an oomycete microorganism that causes a lifethreatening infectious disease, called pythiosis, in humans and animals. The disease has been increasingly reported worldwide. Conventional antifungal drugs are ineffective against P. insidiosum. Treatment of pythiosis requires the extensive removal of infected tissue (i.e., eye and leg), but inadequate surgery and recurrent infection often occur. A more effective treatment is needed for pythiosis patients. Drug repurposing is a promising strategy for the identification of a U.S. Food and Drug Administration-approved drug for the control of P. insidiosum. Disulfiram has been approved to treat alcoholism, but it exhibits antimicrobial activity against various pathogens. In this study, we explored whether disulfiram possesses an anti-P. insidiosum activity. A total of 27 P. insidiosum strains, isolated from various hosts and geographic areas, were susceptible to disulfiram in a dose-dependent manner. The MIC range of disulfiram against P. insidiosum (8 to 32 mg/liter) was in line with that of other pathogens. Proteogenomic analysis indicated that several potential targets of disulfiram (i.e., aldehyde dehydrogenase and urease) were present in P. insidiosum. By homology modeling and molecular docking, disulfiram can bind the putative aldehyde dehydrogenase and urease of P. insidiosum at low energies (i.e., -6.1 and -4.0 Kcal/mol, respectively). Disulfiram diminished the biochemical activities of these enzymes. In conclusion, disulfiram can inhibit the growth of many pathogenic microorganisms, including P. insidiosum. The drug can bind and inactivate multiple proteins of P. insidiosum, which may contribute to its broad antimicrobial property. Drug repurposing of disulfiram could be a new treatment option for pythiosis.

Pythiosis is a difficult-to-treat infectious disease caused by Pythium insidiosum. The condition is unfamiliar among healthcare workers. Manifestation of pythiosis is similar to other fungal infections, leading to misdiagnosis and delayed treatment. The geographical extent of pythiosis at a global scale is unclear. This study aimed to analyze the clinical information recorded in the scientific literature to comprehensively project epidemiological characteristics, clinical features, and future trends of pythiosis. From 1980 to 2021, 4203 cases of pythiosis in humans (n = 771; 18.3%) and animals (primarily horse, dog, and cow; n = 3432; 81.7%), with an average of 103 cases/year, were recruited. Pythiosis case reports significantly increased in the last decade. Pythiosis spanned 23 tropical, subtropical, and temperate countries worldwide. Some patients acquired pythiosis from a trip to an endemic country. Strikingly, 94.3% of human cases were in India and Thailand, while 79.2% of affected animals were in the U.S.A. and Brazil. Clinical features of pythiosis varied. Vascular and ocular pythiosis were only observed in humans, whereas cutaneous/subcutaneous and gastrointestinal infections were predominant in animals. Mortality depended on host species and clinical forms: for example, none in patients with ocular pythiosis, 0.7% in cows with a cutaneous lesion, 26.8% in humans with vascular disease, 86.4% in dogs with gastrointestinal pathology, and 100% in several animals with disseminated infection. In summary, this study reports up-to-date epidemiological and clinical features of pythiosis in humans and animals. It increases awareness of this life-threatening disease, as the illness or outbreak can exist in any country, not limited to the endemic areas.

The fungus-like microorganism Pythium insidiosum causes pythiosis, a life-threatening infectious disease increasingly reported worldwide. Antimicrobial drugs are ineffective. Radical surgery is an essential treatment. Pythiosis can resume post-surgically. Immunotherapy using P. insidiosum antigens (PIA) has emerged as an alternative treatment. This review aims at providing up-to-date information of the immunotherapeutic PIA, with the focus on its history, preparation, clinical application, outcome, mechanism, and recent advances, in order to promote the proper use and future development of this treatment modality. P. insidiosum crude extract is the primary source of immunotherapeutic antigens. Based on 967 documented human and animal (mainly horses) pythiosis cases, PIA immunotherapy reduced disease morbidity and mortality. Concerning clinical outcomes, 19.4% of PIA-immunized human patients succumbed to vascular pythiosis instead of 41.0% in unimmunized cases. PIA immunotherapy may not provide an advantage in a local P. insidiosum infection of the eye. Both PIA-immunized and unimmunized horses with pythiosis showed a similar survival rate of ~70%; however, demands for surgical intervention were much lesser in the immunized cases (22.8% vs. 75.2%). The proposed PIA action involves switching the non-protective T-helper-2 to protective T-helper-1 mediated immunity. By exploring the available P. insidiosum genome data, synthetic peptides, recombinant proteins, and nucleic acids are potential sources of the immunotherapeutic antigens worth investigating. The PIA therapeutic property needs improvement for a better prognosis of pythiosis patients.

AbstrakMedia pembenihan isolasi yang umumnya digunakan adalah media sintetik yang diimpor dan harganya mahal. Banyak bahan organik yang secara alamiah mengalami pemecahan senyawa organik kompleks menjadi senyawa sederhana oleh mikrob. Berdasarkan pemikiran inilah, peneliti mencoba membuat media isolasi dengan bahan dasar limbah air kelapa tua, sehingga dapat menjadi pertimbangan sebagai media yang lebih ekonomis. Metode penelitian adalah eksperimental laboratorik dengan mengisolasi bakteri uji menggunakan media berbahan dasar air kelapa tua. Media air kelapa tua dikomposisikan menyerupai komposisi agar MacConkey dan lempeng agar darah (LAD). Media standar yang digunakan adalah agar MacConkey dan LAD. Media kontrol yang digunakan adalah media agar 15 g/L dan media agar air kelapa tua. Bakteri yang diuji adalah beberapa spesies Enterobacteriaceae dan kokus gram positif. Variabel yang dinilai adalah gambaran makroskopis dan mikroskopis. Analisis data menggunakan teknik Wilcoxon matched pairs test dan sign test. Hasil penelitian menunjukkan perbedaan tidak bermakna (p>0,05) antara media standar dan media air kelapa tua. Gambaran spesifik dari bakteri uji seperti koloni berwarna merah, gambaran mukoid, dan zona hemolitik tampak serupa antara media standar dan media air kelapa tua. Simpulan penelitian ini adalah air kelapa tua dapat dimanfaatkan sebagai bahan dasar media isolasi Enterobacteriaceae dan kokus gram positif. [MKB. 2011;43(3):117-21].Kata kunci: Agar MacConkey, air kelapa tua, isolasi, lempeng agar darah, media Using Ripe Coconut Waste-Water as Base for Isolation Media Substance AbstractThe culture media commonly used for isolation are imported and expensive. Many organic materials are naturally decomposed from complex organic compounds to simple ones by microbes. Based on this principles, this study wants to make isolation media with ripe coconut waste-water as based substance, so it can be considered as economical culture media. The method was laboratoric experimental by isolating tested bacteria with ripe coconut waste-water as based substance. The composition of the media were adjusted with MacConkey agar and blood agar base. Standard media were MacConkey agar and blood agar base. Control media were agar 15 g/L media and ripe coconut waste-water agar media. Tested bacteria were a number species of Enterobacteriaceae and positive gram cocci. The evaluated variables were macroscopic and microscopic images. Data was analized by Wilcoxon matched pairs test and sign test methods. This study did not find a significant differences (p>0.05) between standard media and ripe coconut waste-water media. Specific characteristics of tested bacteria, such as red colonies, mucoid, and hemolitic zone, were similar between standard media and ripe coconut waste-water media. The conclusion is ripe coconut waste-water can be used as base for isolation media substance of Enterobacteriaceae and gram positive cocci. [MKB. 2011;43(3):117-21].

The orphan but highly virulent pathogen Pythium insidiosum causes pythiosis in humans and animals. Surgery is a primary treatment aiming to cure but trading off losing affected organs. Antimicrobial drugs show limited efficacy in treating pythiosis. Alternative drugs effective against the pathogen are needed. In-house drug susceptibility tests (i.e., broth dilution, disc diffusion, and radial growth assays) have been established, some of which adapted the standard protocols (i.e., CLSI M38-A2 and CLSI M51) designed for fungi. Hyphal plug, hyphal suspension, and zoospore are inocula commonly used in the drug susceptibility assessment for P. insidiosum. A side-by-side comparison demonstrated that each method had advantages and limitations. Minimum inhibitory and cidal concentrations of a drug varied depending on the selected method. Material availability, user experience, and organism and drug quantities determined which susceptibility assay should be used. We employed the hyphal plug and a combination of broth dilution and radial growth methods to screen and validate the anti-P. insidiosum activities of several previously reported chemicals, including potassium iodide, triamcinolone acetonide, dimethyl sulfoxide, and ethanol, in which data on their anti-P. insidiosum efficacy are limited. We tested each chemical against 29 genetically diverse isolates of P. insidiosum. These chemicals possessed direct antimicrobial effects on the growth of the pathogen in a dose- and time-dependent manner, suggesting their potential application in pythiosis treatment. Future attempts should focus on standardizing these drug susceptibility methods, such as determining susceptibility/resistant breakpoints, so healthcare workers can confidently interpret a result and select an effective drug against P. insidiosum.

Introduction: Cognitive function is essential for medical students to support learning at the university. One part of cognitive function that is very important in student learning is visual concentration. Visual concentration is the process of maintaining a concentration of thought on a visual object by putting aside other things that are not related. Concentration can be increased in several ways, one of which is to consume nutritious foods that contain flavonoids such as dark chocolate. The purpose of this study was to determine the effect of dark chocolate consumption on visual concentration of cognitive function in Atma Jaya’s medical students.Methods: This was an experimental study and the data were tested using t-paired statistical test. The research was conducted from March to April 2019 at Atma Jaya Catholic University of Indonesia School of Medicine and Health Sciences, Jakarta. The data ovtained from Stroop Test before and after intervention of dark chocolate. A total of 51 respondents were selected according to the research criteria.Results: As many as 48 people (94.1%) experinced an increase in the Stroop Test Score, a person (2%) did not experience a change in the Stroop Test Score, and two people (3.9%) experinced a decrease in the Stroop Test Score. The result of statistical test show significant results (p<0.01). The average score before the intervenion was obtained at 77.02±10.42 and the average score after the intervention was obtained at 77.02±11.81 (Δ= +12.26).Conclusion: There is an effect of dark chocolate consumption on visual concentration of cognitive function in Atma Jaya’s medical student

Background <br />The prevalence of type 2 diabetes mellitus (DM) is increasing. Diabetic patients have a higher risk of getting dermatomycosis. Dermatomycoses, although a common health problem amongst DM, is often misdiagnosed and consequently undertreated. Studies on the association between dermatomycosis and type 2 diabetes are lacking, especially in Indonesia. Therefore, the aim of this study was to determine the prevalence, etiology, and association of dermatomycosis with diabetic control of type 2 DM. <br /><br />Methods<br />A cross-sectional study was performed involving 87 subjects with type 2 DM. Demographic and clinical data, including age, sex, and blood glucose level, were collected. If a dermatomycosis lesion was found, a specimen would be taken for identification. Determination of serum glucose level was conducted using Roche c111 analyzer®. Statistical analysis was performed with the chi-square test and Kolmogorov-Smirnov two-independent sample test.<br /><br />Results<br />Seventeen (19.55%) subjects had dermatomycosis. The predominant age group affected was 51 - 60 years (42.4%). The number of clinically apparent dermatomycosis was greater in the uncontrolled than in the controlled blood sugar group, but the difference was statistically not significant (p &gt; 0.05). The lesions were mostly found on the nails (74%) and the most common etiology was candida (50%) followed by dermatophyte (25%) and non-dermatophyte molds (25%). <br /><br />Conclusion<br />Uncontrolled blood sugar tends to increase the risk of dermatomycosis in type 2 DM patients. Fungal skin infections are common in type-2 DM patients, especially in those with poor glycemic control.