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Loeki Enggar Fitri,1,2 Aulia Rahmi Pawestri,1,2 Nuning Winaris,1,2 Agustina Tri Endharti,1 Alif Raudhah Husnul Khotimah,3,4 Hafshah Yasmina Abidah,3,4 John Thomas Rayhan Huwae3,5 1Department of Parasitology Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia; 2AIDS, Toxoplasma, Opportunistic Disease and Malaria Research Group, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia; 3Master Program in Biomedical Science, Faculty of Medicine, Universitas Brawijaya, Malang, Indonesia; 4Medical Doctor Profession Education, Faculty of Medical and Health Science, Maulana Malik Ibrahim State Islamic University, Malang, Indonesia; 5Medical Doctor Profession Study Program Faculty of Medicine, Universitas Brawijaya, Malang, IndonesiaCorrespondence: Alif Raudhah Husnul Khotimah, Master Program in Biomedical Science, Faculty of Medicine, Universitas Brawijaya, 65145, District Lowokwaru, Malang, East Java, Indonesia, Tel +62 822 573 458 27, Fax +62 341 564755, Email husnulk556@gmail.com; alifraudhah_hk@student.ub.ac.idAbstract: Malaria remains to be a national and global challenge and priority, as stated in the strategic plan of the Indonesian Ministry of Health and Sustainable Development Goals. In Indonesia, it is targeted that malaria elimination can be achieved by 2030. Unfortunately, the development and spread of antimalarial resistance inflicts a significant risk to the national malaria control programs which can lead to increased malaria morbidity and mortality. In Indonesia, resistance to widely used antimalarial drugs has been reported in two human species, Plasmodium falciparum and Plasmodium vivax. With the exception of artemisinin, resistance has surfaced towards all classes of antimalarial drugs. Initially, chloroquine, sulfadoxine-pyrimethamine, and primaquine were the most widely used antimalarial drugs. Regrettably, improper use has supported the robust spread of their resistance. Chloroquine resistance was first reported in 1974, while sulfadoxine-pyrimethamine emerged in 1979. Twenty years later, most provinces had declared treatment failures of both drugs. Molecular epidemiology suggested that variations in pfmdr1 and pfcrt genes were associated with chloroquine resistance, while dhfr and dhps genes were correlated with sulfadoxine-pyrimethamine resistance. Additionally, G453W, V454C and E455K of pfk13 genes appeared to be early warning sign to artemisinin resistance. Here, we reported mechanisms of antimalarial drugs and their development of resistance. This insight could provide awareness toward designing future treatment guidelines and control programs in Indonesia.Graphical Abstract: Keywords: Indonesia, antimalarial resistance, chloroquine, sulfadoxine-pyrimethamine, primaquine, artemisinin

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.

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.

This study was conducted cross sectionally in Maumere, Sikka Regency, East Nusa Tenggara Province, that previously known as one of malaria endemic area with high incidence of low birth weight (LBW) cases. In this study the peripheral and umbilical blood samples as well as placental tissues were collected from mother delivered their babies with LBW at the TC Hiller Regional Hospital. All of the blood samples were examined for the presence of parasites by microscopic and PCR technique, while the plasma levels of VEGF, PlGF, and VEGFR-1, VEGFR-2, and HIF 1-�� were determined using ELISA. The sequestration of infected erythrocytes and hemozoin were determined from placental histological slides and the expression of placenta angiogenic factors were observed by immunofluorescent technique.

Malaria infection in pregnant women can cause various clinical manifestations, one of them is fetus low birth weight. Sequestrations of infected erythrocytes and macrophages on intervillous space has been revealed by previous study, however the mechanism of this sequestration in placenta insufficiency is still unknown. We suggested that local immunity on placenta tissue can induce the production of reactive oxygen species (ROS) and reactive nitrogen species (RNS) that can cause the destructions of placenta tissue due to the fetus low birth weight. This research was conducted to explore the path of immunity response on malaria placenta which involved the expression of inducible nitric oxide synthase (iNOS) and the level of malondialdehyde (MDA). This experimental laboratory study was done in pregnant Balb/c strain mice that infected withPlasmodium bergheiin 9thday of pregnancy. Eighteen mice were divided into control group (pregnant mice) and treatment group (pregnant mice infected withPlasmodium berghei). iNOS expression was detected by using polyclonal antibody t iNOS, and level of MDA in the placenta tissue was measured by NTB assay. Fetal birth weight was measured on 15thday of pregnancy using analytic balance. The t-test independent showed a significant difference on iNOS expression between control group and treatment group (p

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).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.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; P0.001).Although microscopy remains the most appropriate method for the diagnosis of CM in remote areas, nPCR can be considered a complementary test.

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 rRNA by polymerase chain reaction showed the presence of P. vivax and absence of P. falciparum. All patients responded well to antimalarial drugs.