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This study aimed to obtain a microbial active compound as a novel antimalarial drug from Indonesian isolates. Target-based assays were used to screen for antimalarial activity against the parasite mitochondrial, Plasmodium falciparum malate:quinone oxidoreductase (PfMQO) enzyme. In total, 1600 crude extracts, composed from 800 fungi and 800 actinomycetes extracts, were screened against PfMQO, yielding six active extracts as primary hits. After several stages of stability tests, one extract produced by Aspergillus sp. BioMCC f.T.8501 demonstrated stable PfMQO inhibitory activity. Several purification stages, including OCC, TLC, and HPLC, were performed to obtain bioactive compounds from this active extract. All purification steps were followed by an assay against PfMQO. We identified the active compound as nornidulin based on its LC-MS and UV spectrum data. Nornidulin inhibited PfMQO activity at IC50 of 51 µM and P. falciparum 3D7 proliferation in vitro at IC50 of 44.6 µM, however, it had no effect on the growth of several mammalian cells. In conclusion, we isolated nornidulin from Indonesian Aspergillus sp. BioMCC f.T.8501 as a novel inhibitor of PfMQO, which showed inhibitory activity against the proliferation of P. falciparum 3D7 in vitro. [ABSTRACT FROM AUTHOR]

Background: In Drug-Resistant Tuberculosis (DR-TB), high IL17 level was corelated with severe tissue damage while low level was associated with mortality. High levels of IL-17 and IL - 23 were found in patients with a high bacterial load. TB treatment is carried out by combination of anti-tuberculosis drugs for a period of 9 to 24 months. This study aimed to analyze the differences in IL-17 and IL-23 levels before and after treatment compared to healthy controls, and their relationship with conversion AFB smear. Methods: A prospective cohort study was conducted with total 38 subjects diagnosed as DR-TB using a molecular rapid test and 34 normal subjects as case and control groups. Examination of sputum AFB, IL-17 and IL-23 levels were carried out at the start of treatment and 30 days after treatment. Treatment was given in short term and individul drugs resistant regiments. Results: The average levels of IL-17 and IL 23 before treatment compared to those after treatment showed no significant difference (p=0.153; p= 0.328). When compare to healthy controls group, the mean levels of IL-17 and IL 23 before treatment was not significantly different (p= 0.883; p= 0.928), whereas after 1 month treatment were significantly different (p= 0.047; p=0.042). An increase of IL-17 levels followed by an increase in IL-23 levels (p=0,000 r=.0.872). TB treatment affected the conversion rate of 97.4% with linear association. Conclusion: The levels of IL-17 and IL-23 after treatment were increasing and significantly different with healthy group. These related with the high AFB conversion rate. [ABSTRACT FROM AUTHOR]

Introduction: Previous research has proven that ethyl acetate extract of Streptomyces hygroscopicus can inhibit Plasmodium growth in vivo and in vitro through inhibition of the Proteasome ubiquitin system. In this study, we analyzed the activity of several fractions of S. hygroscopicus toward the growth of P. falciparum culture with specific targets on the mitochondrial enzyme of P. falciparum; L-malate: quinone oxidoreductase (PfMQO) and dihydroorotate dehydrogenase (PfDHODH). Methodology: the fractionation process uses the Flash Column Chromography (FCC) BUCHI Reveleris PREP, analyzed using thin layer chromatography (TLC). The antimalarial activity of the fractions was tested on the culture of P. falciparum 3D7 using enzyme lactate dehydrogenase (PfLDH) assay and their activity toward PfMQO and PfDHODH target enzymes. The fractions analyzed using HPLC and compared to the standard dihydroeponemycin. The toxicity of the fractions, the WST-8 assay was conducted. Results: of the 30 fractions produced, 6 fractions were able to suppress the P. falciparum and two of them were able to inhibit more than 50% of P. falciparum growth. Both of these fractions were proven to inhibit the activity of the PfMQO enzyme but had no inhibition of the PfDHODH enzyme. Both fractions reduced human cell viability by less than 50% in toxicity test. HPLC analysis indicated that besides dehydroeponemycin, there were other compounds presences in the extract. Conclusions: fractions of S. hygroscopicus metabolite contains non-toxic compounds in addition to dihydroeponemycin that could suppress the growth of P. falciparum in vitro. This potential effect might be related to the inhibition of PfMQO enzyme. [ABSTRACT FROM AUTHOR]

Background: Malaria eradication has been complicated by the repeated emergence of antimalarial drug resistances. We aimed to determine whether a metabolite extract of Streptomyces hygrocopicus subsp. hygroscopicus could decrease the viability of Plasmodium falciparum 3D7 in vitro. Methods: S. hygroscopicus subsp. hygroscopicus isolates were inoculated and fermented on the ISP4 medium. The fermented S. hygroscopicus was mixed with ethylacetate 1:5 (v/v), and the solvent phase was evaporated. Several concentrations of isolated extract was added to the P. falciparum 3D7 culture containing trophozoite and schizont stages in 24 wells plates when the degree of parasite-infected erythrocytes reached 5%, then incubated for 8 hours. DNA parasite density was measured using flow cytometry, parasite degree and morphology were observed under microscopic by Giemsa-stained smears. Results: The metabolite extract affected the morphology of almost all of parasite asexual stages. Schizonts and trophozoites failed to grow and appeared damaged with pycnotic cores and loss of cytoplasmic content. At 8 hours there was a significant decrease in DNA parasite density in culture exposed to 2.6 mg/ml and 13 mg/ml (P = 0.002; P = 0.024) of the extract. The degree of parasite-infected erythrocytes was decreased from the beginning of exposure (0.02 mg/ml of the extract). There was a significant inverse correlation between the concentration of extract and the degree of parasite-infected erythrocytes as well as the density of DNA parasite (r = −0.772, P = 0.000; r =−0.753; P =0.000). Conclusion: Metabolite extract of S. hygroscopicus subsp. hygroscopicus causes morphological damage, decreases the degree of parasite-infected erythrocytes and the DNA density of P. falciparum 3D7 in vitro. [ABSTRACT FROM AUTHOR]

Malaria is a disease caused by Plasmodium infection. Of the 5 species that infect humans, Plasmodium vivax and Plasmodium falciparum are the two most common species found in Indonesia. Early and accurate diagnosis is crucial for effective treatment and disease management. Conventional diagnostic methods, microscopy, and rapid diagnostic tests (RDTs) have limitations in sensitivity and specificity. PCR-based assays have undergone significant enhancement such as the development of touchdown PCR and primer designing. However, the use of PCR-based assays in Plasmodium vivax infections still has several limitations including the formation of dimers and cross reactivity. Two isolated Plasmodium vivax DNA samples taken from malaria confirmed patients and stored in the Biomedical Laboratory, Faculty of Medicine, Universitas Brawijaya were tested with both standard Plasmodium vivax (rViv) primers and new (Pv-Rv) primer for Plasmodium vivax 18SSU RNA gene detection. Each pairs of primers underwent both regular nest 2-PCR and touchdown nest 2-PCR. The PCR products were checked by 3% agarose gel electrophoresis, and then quantified by using GelDoc Imaging System and ImageJ software. The paired t-test was used to analyze the results statistically. The results showed that, using rViv primers, there was no statistically significant difference in the ratio of band intensity to background intensity between regular nest 2-PCR and touchdown nest 2-PCR (P > 0.05). However, when the background intensity was analyzed, a statistically significant difference was discovered (P < 0.05). By using Pv-Rv primers, a significant difference in the background intensity as well as in the ratio of band intensity to background intensity between regular nest 2-PCR and touchdown nest 2-PCR was observed (P < 0.05). The results indicated that application of touchdown nest 2-PCR had an effect in reducing background noise by using either rViv primers (standard primer) or Pv-Rv primers (new primer). Further research and modification of Pv-Rv primers were required for Plasmodium vivax identification. [ABSTRACT FROM AUTHOR]

Background: Toxoplasma gondii is one of the zoonotic protozoa parasites. It can prevalently infect humans and warmblooded animals, causing human health problems and substantial economic losses to the livestock industry worldwide. Chicken is one of the potential sources of toxoplasmosis, but there is no report of the prevalence of toxoplasmosis and their genotypes in free-range chickens in Libya. Aim: This study aims to conduct a survey of molecular prevalence and identify the T. gondii genotype in free-range chickens and its association with the risk factors of age, gender, and region in Northeastern Libya. Methods: This study was conducted by examining a total of 315 free-range chicken organs (brain and heart) derived from three administrative districts in Northeastern Libya. The molecular prevalence was determined by PCR technique using B1 gene amplification. and the T. gondii genotype was determined by nested PCR-RFLP of GRA6 gene amplicon with restriction enzymes (MseI). Results: The overall molecular prevalence of T. gondii in free-range chicken in all three districts was 9.5% (30/315), and the highest (15.4%) was in the Al-Marj district (p = 0.01; x² = 9.238). The highest prevalence of T. gondii by age was in chickens aged more than 2 years (p = 0.001; x² = 15.530). The difference in T. gondii prevalence in male and female chickens was not significant (p = 0.372; x² = 0.798). The predominant genotype I (93.3%) had identified at position 544 and 194 bp at the GRA6 marker, and only two positives were from genotype II (6.7%) at 700 and 100 bp fragments. Conclusion: The molecular prevalence of toxoplasmosis in free-range chicken in three districts in Northeastern Libya was 9.5%, and the highest rate was shown in the Al Marj district. Chicken by age more than 2 years had more risk to transmit toxoplasmosis in human. There was no different infection risk by consuming male or female free-range chicken. It is the first report to determine the predominant genotype, which was genotype I. [ABSTRACT FROM AUTHOR]

Most of the complications of malaria such as anaemia, thrombocytopenia, jaundice, and renal failure are commonly found in Plasmodium falciparum malaria, but the incidence of severe and complicated vivax malaria tends to be increasing. We report two cases of severe Plasmodium vivax malaria from Malang, a nonendemic area in Indonesia. Patients exhibited anaemia, thrombocytopenia, jaundice, renal disturbance, and melena. Microscopic peripheral blood examination and amplification of parasite 18s rRNAby polymerase chain reaction showed the presence of P. vivax and absence of P. falciparum.All patients responded well to antimalarial drugs. [ABSTRACT FROM AUTHOR]

Introduction. The imbalance of the immune response is an important factor contributing to the incidence of ocular toxoplasmosis (OT). Regulatory T cells (Treg) play a key role in maintaining the balance between Th1 and Th17 immune responses, while interleukin-27 (IL-27) levels are related to the differentiation of Th17 cells. This study analyzes the differences in the number of Treg cells and the level of IL-27 between OT patients and seropositive individuals without ocular lesions and its correlation with retinal lesion size. Methods. This analytic observational study, conducted for 8 months, involved 11 OT patients and 10 seropositive individuals without ocular lesions. All subjects underwent a comprehensive ophthalmological examination. Retinal lesions were documented by fundus photographs and the size was measured using Digimizer 4.2.2.0 software. Isolation of peripheral blood mononuclear cells (PBMC) was performed to measure the number of Treg cells using flow cytometry and interleukin-27 levels were assessed using the Sandwich enzyme-linked immunosorbent assay (ELISA) technique. Data were analyzed with SPSS. Result. The number of Treg cells in the OT group (47.16 ± 15.66%) was lower than in the seropositive group without the ocular lesions (62.86 ± 17.08%) (p = 0.029). The serum IL-27 levels in the OT group were not significantly different from the seropositive group without the ocular lesions (p = 0.360). The number of Treg cells was significantly related to retinal lesion size (p = 0.043), with a correlation coefficient of −0.648, indicating a strong and inverse correlation. There was no significant correlation between serum IL-27 levels and retinal lesion size (p = 0.556). Conclusion. Ocular toxoplasmosis patients have a low number of Treg cells that are inversely related to the retinal lesion size. The size of the retinal lesion increases as the number of Treg cells decreases. [ABSTRACT FROM AUTHOR]

The article discusses a study on the diagnostic difficulties with Plasmodium knowlesi infection in humans which is considered the fifth human pathogenic malaria parasite. Results of the testing of blood samples from gold miners with uncomplicated malaria indicated cases of Plasmodium knowlesi while DNA sequencing confirmed the presence of Plasmodium vivax infections. Study results also showed the geographic distribution of natural Plasmodium knowlesi human infections.

The high mortality and morbidity rates of malaria remains to be a global problem, particularly in tropical regions. Cerebral malaria, the leading cause of death, possesses a complex pathogenesis involving host immune and parasite interactions. Lately, according to the gut-brain axis concept, the gut microbiota was identified to protect the host against systemic pathogen infection through the modulation of immune signaling pathways. In this study, we aimed to explore the effect of probiotic supplementation toward neurological outcomes of cerebral malaria in vivo. Twenty-eight male C57BL/6 mice were randomly divided into 5 groups after 2 weeks acclimatization period. All groups except the negative control were given oral probiotic supplementation with either Lactobacillus casei at 109 colony forming unit/100 µL (CFU/100 µL), Bifidobacterium longum at 109 CFU/100 µL, or the combination of each at 5x108 CFU/100 µL for five days before to six days after Plasmodium berghei infection. Neurological manifestations were recorded by using the SmithKline, Harwell, Imperial College, Royal Hospital, Phenotype Assessment (SHIRPA) score. The data was analyzed by using One-way ANOVA and post-hoc LSD. The results showed that there were significant differences of motor behavior (P < 0.01), muscle tone (P < 0.01), reflex and sensory (P < 0.05), and neuropsychiatric state (P < 0.05) between the control group and that receiving combination of L. casei and B. longum. The results suggested that probiotic supplementation showed promising effects in improving the neurological manifestations during cerebral malaria. Further studies are required to explicate the pathways involved in the immunological interplay. In the future, it is propitious that the mortalities by cerebral malaria could be mitigated through the optimal use of probiotics. [ABSTRACT FROM AUTHOR]

The mucosal immune system contributes for the largest component of the tissue immune system due to its massive surface area and constant exposure to the microbiota. The gut microbiota comprises a complex micro‐ecosystem in the intestine and plays a major role in regulating innate and adaptive immunity. Several studies revealed that infectious diseases involve bidirectional interactions in the gut microenvironment, including changes in the gut microbiota composition. During Plasmodium infection, an increase of pro‐inflammatory cells in the lamina propria and a shift in the composition of the gut microbiota contribute to intestinal ecosystem dysbiosis. Although the mechanisms of this dysbiosis is still uncertain, it is thought to be associated with the sequestration of infected red blood cells in the intestinal microvascular system, leading to endothelial villous disruption, and thus activating effector immune cells scattered in the intestinal epithelium and lamina propria. This review provides information on this conjoint interaction which will be beneficial to modulate the host immune response in malaria through manipulation of the gut microbiota composition. [ABSTRACT FROM AUTHOR]

Background and Aim: Zebrafish are frequently used as model organisms in scientific research as their genes mirror those of humans. Aeromonas hydrophila bacteria can infect humans and animals, mainly fish. This study aimed to identify the concentration and route of A. hydrophila infection in adult zebrafish. Zebrafish had been used as a challenge test by analyzing their hematological profiles, blood glucose levels, and survival rates. Materials and Methods: Induction of cell supernatant free (CSF) from A. hydrophila bacteria in adult zebrafish was carried out via bath immersion (BI), intraperitoneal injection (IPI), intramuscular injection (IMI), and healthy zebrafish as a control (C). The bacterial concentrations were 107, 109, and 1011 colony-forming units (CFU)/mL. At 24 h post-infection, the outcomes of infection were evaluated based on survival rates, hematological profiles, and blood glucose levels. A oneway analysis of variance with a confidence level of 95% was employed to examine the data. Results: In the BI, IPI, and IMI treatment groups, the survival rate of the fish reached a peak of 100%, 22%-100%, and 16%-63%, respectively, compared with the injection technique. In the IMI2 group, a 109 CFU/mL bacterial concentration was determined to correspond to the lethal dosage 50. All infection groups had lower erythrocyte and hemoglobin counts but higher leukocyte counts than the control group. The blood sugar levels of the healthy and infected groups were not significantly different. Conclusion: The route of A. hydrophila infection through Intramuscular injection with a concentration of 109 CFU/mL indicated a high performance compared to other techniques. This method could be developed as a reproducible challenge test. [ABSTRACT FROM AUTHOR]

Background: Staphylococcus and Aeromonas bacteria are pathogens in humans and animals. The therapy disrupts the virulence structure of the bacteria, resulting in bacterial death. Currently, chemical drugs have resulted in many resistant bacteria, so it is necessary to find alternative natural materials that are not toxic and do not quickly induce resistance. Aims: This study aimed to analyze the potential of methanol extract from Black soldier fly (BSF) prepupae as an antibacterial agent against Staphylococcus aureus and Aeromonas through in silico and in vitro tests. Methods: The BSF prepupae methanol extract was analyzed for protein and fatty acid contents. Disc diffusion method, minimal inhibitory concentration, and minimum bactericidal concentration test were used for in vitro tests against Staphylococcis and Aeromonas. Molecular docking of the active ingredients (defensin, chitin, and chitosan as well as fatty acids) in BSF was downloaded from the NCBI database and docked by the Hex Cuda version 8.0 program with Correlation type parameters Shape + Electro and Grid Dimension version 0.6. Docking results were analyzed using the Discovery Studio program version 21.1.1. Results: The highest fatty acid contents in the extract were palmitic acid and myristic acid. Methanol extract from BSF prepupae acted as a bactericidal agent against S. aureus at a concentration of 320 mg/ml, in contrast to Aeromonas, which still showed bacterial growth. The results of the in silico test showed that defensin--aerolysin and defensin-- hemolysin was bound to the same active site area. However, the amount of binding energy produced by 69-Defensin- 83-aerolysin was higher than all defensin types in BSF against Aeromonas. Chitin and chitosan showed a bond on the active site of aerolysin and hemolysin, but chitosan had a stronger bond than chitin. In silico study also showed the strongest binding affinity of BSF fatty acids to isoleucyl-tRNA synthetase of S. aureus. Conclusion: The study showed that methanol extract from BSF prepupae had potential capability as an antibacterial agent against S. aureus than Aeromonas in vitro and in silico. [ABSTRACT FROM AUTHOR]

Background: Malaria screening for blood derived from any donors prior to transfusions is a standard procedure that should be performed; but, in fact, it is not routinely conducted. In case of the blood is infected with Plasmodium spp., the survival of parasites may be depending on, or even influencing, the profile of red blood cells (RBCs). Methods: This observational longitudinal study was conducted upon 55 bags of donor blood that randomly selected. Malaria infections were detected using Rapid Diagnostic Test/RDT with thin and thick blood smear confirmation. The changes of Plasmodium spp. viability and RBCs profiles, as well as other hematological parameters, were observed from the results of routine hematological examinations which were performed on days 1,7,14 and 21 of storage. Results: Among 55 blood samples, there were 17 and 38 bags, respectively, positive and negative for malaria, then used for analysis as the case and control groups. There were significant decreasing values (p< 0.05) of all routine blood examination parameters of donor blood, started from days 1, 7, 14, 21, and 28. There were no differences in decreasing profiles between those infected and non-infected donor blood (p> 0.05). On days 21 and 28 none of the positive samples still contained parasites. Conclusion: Erythrocytes profiles of donor blood significantly decreased with the duration of storage, but were not influenced by the presence of Plasmodium spp. [ABSTRACT FROM AUTHOR]

Background: We aimed to reveal the role of CD11b and hypoxia-inducible factors- 1alpha (HIF-1α) expressions on monocytes and alveolar macrophages of lung tissue, and the levels of serum surfactant protein-D (SP-D) in severe malaria-associated acute lung injury (ALI). Methods: The C57BL/6 mice were divided into control group, renal malaria group (inoculated with 106 Plasmodium berghei ANKA), and cerebral malaria group (inoculated with 107 P. berghei ANKA). The expressions of CD11b and HIF-1α in lung tissue were observed by immunohistochemistry, and serum SP-D levels were measured by ELISA. This study was conducted from June 2014 to February 2015 in the Laboratory of Parasitology, Faculty of Medicine, Universitas Brawijaya, Malang. Results: The CD11b expression on pulmonary tissue of renal and cerebral malaria mice were significantly higher than control mice (P=0.002; P=0.002), as well as the HIF-1α expression on pulmonary tissue (P=0.002; P=0.002). The level of serum SP-D in renal malaria and cerebral malaria mice were significantly higher than control mice (P=0.002; P=0.002). We found a strong correlation between the expression of CD11b and HIF-1α in lung tissue (r=0.937, P=0.000), as well as between CD11b expression and serum SP-D levels (r=0.907, P=0.000) and between HIF-1α expression and serum SP-D levels (r=0.913, P=0.000). Conclusion: Severe malaria-associated ALI increased the expression of CD11b and HIF-1α in the lung tissue and increased serum SP-D levels of C57BL/6 mice significantly. [ABSTRACT FROM AUTHOR]

Background: During pregnancy, the balanced dominance of the T helper17 response shifts to a Th2 response that is characterised by the production of IL-10, following the completion of the implantation process. Transforming growth factor-β (TGF-β) expression is associated with the completion of trophoblast invasion and placental growth. This study assessed the effect of malaria infection on the levels of IL-17, IL-10, and TGF-β in the plasma of pregnant mice with malaria. Methods: Seventeen pregnant BALB/C mice were divided into two groups: mice infected with Plasmodium berghei (treatment group) and uninfected mice (control group). The mice were sacrificed on day 18 post-mating. Parasitemia was measured by Giemsa staining. The levels of IL-17, IL-10, and TGF-β were measured by ELISA. Results: Using independent t test, the IL-17 levels in the treatment group were higher than those in the control group (P = 0.040). The IL-10 levels in the treatment group were lower than those in the control group (P = 0.00). There was no significant difference in the TGF-β levels (P = 0.055) between two groups. However, using SEM analysis the degree of parasitemia decreased the plasma TGF-β levels (tcount = 5.148; ≥ ttable= 1.96). SEM analysis showed that a high degree of parasitemia increased the IL-17 levels and decreased the IL-10 and TGF-β levels. Conclusion: Malaria infection during pregnancy interferes with the systemic balance by increasing the IL-17 levels and decreasing the IL-10 and TGF-β levels. [ABSTRACT FROM AUTHOR]

Background: The innate immune response to tuberculosis infection may involve the increased production of nitric oxide and cathelicidin due to the up-regulated expression of the vitamin D receptor (VDR), though this proposed mechanism remains controversial. The aim of this study was to determine how the exposure of human monocytes to Mycobacterium tuberculosis (M. tuberculosis) DNA affects the production of nitric oxide and cathelicidin, as well as the expression of VDR. Methods: This study was performed using monocytes obtained from healthy donors. After 24 h incubation, monocytes were stimulated with M. tuberculosis DNA for 18 h to determine the expression of VDR mRNA and the production of nitric oxide and cathelicidin versus non-stimulated cells (the control group). Results: The expression of VDR mRNA was higher in the monocytes exposed to M. tuberculosis DNA compared to the control group (P = 0.020). Monocytes exposed to M. tuberculosis DNA also showed significantly increased production of nitric oxide and cathelicidin compared to the control group (P = 0.0001; P = 0.028). Conclusion: The stimulation of human monocytes with M. tuberculosis DNA increases the expression of the VDR mRNA and the production of nitric oxide and cathelicidin. [ABSTRACT FROM AUTHOR]

Background: Microscopic detection is the conventional method for detecting malaria parasites. Although it is efficient and inexpensive, it has its limitations. In recent years, polymeras chain reaction (PCR) has been considered superior to microscopy in detecting mixed Plasmodium infections or infections with low parasite density. To determine whether microscopic or nested PCR (nPCR) is better at detecting congenital malaria (CM). Methods: Blood smear examination and nPCR were performed with blood samples taken from mothers and their newborns, who were likely to be suffering from CM and in whom one of the symptoms was low birth weight (LBW). The sensitivity and specificity of each method were then compared. Results: During one year of study, the prevalence of CM among 92 LBW newborns was determined to be 6.8% using microscopy and 7.8% using nPCR. Among the 92 mother-infant paired subjects, CM was detected in 34 subjects (37%) by microscopy and in 39 subjects (42.4%) by nPCR. nPCR was more sensitive (76.5% vs 66.7%) but less specific (77.6% vs 84.9%) than microscopy. When the two methods were compared, nPCR gave significantly better results in diagnosing CM (AUC = 0.770; P < 0.001). Conclusion: Although microscopy remains the most appropriate method for the diagnosis of CM in remote areas, nPCR can be considered a complementary test. [ABSTRACT FROM AUTHOR]