Genetic Variability of the Long-Tailed Macaque (Macaca fascicularis) Populations in Urban Habitat in Padang City, West Sumatra, Indonesia

The long-tailed macaque (Macaca fascicularis) is a primate species recognized for its exceptional ability to adapt to urban habitat. However, urban anthropogenic activities contribute to the fragmentation of macaque natural habitat, affecting genetic variation among distinct populations. Therefore, this study aimed to assess the genetic variability of M. fascicularis populations in Padang City, West Sumatra, Indonesia. A total of 70 fecal samples from the wild long-tailed macaques in Gunung Padang (GPD), Gunung Meru (GMR), and Gunung Pangilun (GPG) were collected using a non-invasive method. Conventional PCR amplification and Sanger sequencing were conducted to examine a 1,200-bp mitochondrial DNA (mtDNA) fragment in the D-loop region. The analysis of genetic variation showed that only two haplotypes were present in the three populations. Both GPD and GMR shared the same haplotype (H1), while the GPG population had a distinct haplotype (H2). No intrapopulation variation was observed, and haplotype differences were found in ten nucleotide sites with transition substitution mutations. These results showed limited genetic variation among populations of the long-tailed macaque in Padang, thereby providing valuable insights for stakeholders when formulating genetic conservation policies

PEMBELAJARAN PENDIDIKAN AGAMA ISLAM DAN BUDI PEKERTI DI ISLAMIC BOARDING SCHOOL

This study aims to describe the implementation of Islamic Religious Education and Character in Islamic Boarding schools. The method used for this research is a qualitative method with a library research design. Whereas for the research instruments, data collecting is documentation and observation. Based on research results shows that the implementation of Islamic Religious Education and Character in Islamic Boarding School is based on the national curriculum.  The learning method used is the classical learning method, such as the lecture method, the question-answer method, and the storytelling method. Meanwhile, the modern learning method is active learning, such as the PAIKEM model

Ekspresi Enzim Rekombinan Reverse Transcriptase (RTRNase H) Simian Betaretrovirus Serotipe-2 Asal Macaca fascicularis Indonesia dalam Sistem Ekspresi Eschericia coli

Reverse transcriptase (RT) enzyme is a key tool in molecular biology for the synthesis of complementary DNA (cDNA) from messenger RNA (mRNA). Combining RT activity with PCR amplification has been a gold standard as the first step in cloning the coding region of any gene of interest. Evidently, RTs have been critical in advancing molecular biology, genetics and medicine to their current stage. In this study, we were developing the RTDRNase H recombinant enzyme isolated from serotype-2 simian betaretrovirus-2 (SRV-2) infected Indonesian Macaca fascicularis using Escherichia coli expression system. The study was conducted using RT SRV-2 gene expression using E. coli expression system, proteins purification, and application to RT PCR technique. The SDS PAGE expression analysis showed a specific band size of 32.7 kDa assumed as RT protein enzyme. Application of RT SRV-2 enzyme generated to the RT PCR technique of β-globin CDV and SRV-2 env gene target showed that the RT SRV-2 enzyme was capable to reverse transcribed mRNA into cDNA as indicated by the presence of specific DNA band compared with commercial RT enzymes. This RT SRV-2 enzyme showed its activity similar to that of commercial one, although the activity was lower

Identification and molecular characterization of simian endogenous retrovirus in Macaca fascicularis and Macaca nemestrina from captive breeding facilities in Bogor, Indonesia

Background and Aim: Endogenous retroviruses (ERVs) found in all vertebrates, including non-human primates (NHPs), are known to be genetically inherited. Thus, recent studies have explored ERVs for human immunodeficiency virus vaccine development using human ERV (HERV) due to the hypervariability of exogenous retroviruses which cause conventional vaccines to be ineffective. HERV was also found to be able to induce an immune response in cancer patients. This study aimed to identify and molecularly characterize ERVs from Indonesian NHPs: Macaca fascicularis and Macaca nemestrina. Then, we described the phylogenetic relationship of these isolates with those of the simian ERVs (SERVs) characterized in other species and countries. Materials and Methods: First, 5 mL of whole blood samples was taken from 131 long-tailed macaques and 58 pig-tailed macaques in captive breeding facilities at Bogor, Indonesia, for DNA extraction. Next, the DNA samples were screened using the SYBR Green real-time polymerase chain reaction (PCR) technique with specific primers for env (simian retroviruses [SRV]1-5 7585U19 and SRV1-5 7695L21). Positive SERV results were those with cycle threshold (CT) values < 24 (CT < 24) and melting temperature (TM) ranges of 80°C–82°C. Then, whole-genome nucleotide sequences from two pig-tailed macaques samples detected as positive SERV were generated using a nucleic acid sequencing technique which utilized the walking primer method. Subsequently, the sequences were analyzed using bioinformatics programs, such as 4Peaks, Clustal Omega, and BLAST (NCBI). Subsequently, a phylogenetic tree was constructed using the neighbor-joining method in MEGA X. Results: SYBR Green real-time PCR amplification results indicated that SERV (Mn B1 and Mn B140910)-positive samples had CT values of 22.37–22.54 and TM of 82°C. Moreover, whole-genome sequences resulted in 7991 nucleotide sequences, comprising long terminal repeat, gag, pro, pol, and env genes identical between the sequenced samples. Furthermore, the phylogenetic tree results indicated that both samples from M. nemestrina had 99%–100% nucleotide identities to the Mn 92227 sample identified at the National Primate Center University of Washington (NaPRC UW) which was imported from Indonesia in 1998, confirmed as a novel SERV strain. The phylogenetic tree results also indicated that although SERV whole-genome nucleotide and env amino acid sequences were clustered with SRV-2 (identity values of 82% and 79%, respectively), they had a 99%–100% nucleotide identity to Mn 92227. Meanwhile, the gag, pro, and pol amino acids were clustered with SRV-1, SRV-3, SRV-4, SRV-5, SRV-8, and SERV/1997, with 82% and 88% identity values. Conclusion: Based on the SYBR Green real-time PCR profiles generated, similarities with Mn 92227 were observed. Subsequent phylogenetic analysis confirmed that both samples (Mn B1 and Mn B140919) from pig-tailed macaques in the country of origin were novel SERV strains at NaPRC UW. Therefore, it could be used in biomedical research on ERVs

Expression of SARS-CoV-2 Nucleocapsid (N) Recombinant Protein Using Escherichia coli System

One of the main antigen that can be used for serological testing is the nucleocapsid (N) which is the most abundant viral-derived protein in SARS-CoV-2 where this virus can cause COVID19 disease. The aim of this study was to develop the SARS-CoV-2 N recombinant protein using Escherichia coli expression system. A total of 1,089 nucleotides encoding 362 amino acids of SARS-CoV-2 N was cloned to pET-14b vector. The plasmid then expressed in E. coli BL21 (DE3) and induced with 1.0 mM IPTG (Isopropyl-β-d-1-thiogalactopyranoside). The cell was harvested using denaturation lysis buffer due to inclusion body formation of SARS-CoV-2 N protein. Dialysis processed and concentrated using PEG-6000 resulted 0.992 mg/ml protein yield. Analysis of SARS-CoV-2 N recombinant protein using SDS-PAGE technique showed approximately 37.0 kDa specific band target protein. Application of this SARS-CoV-2 N recombinant protein to vaccinated and non-vaccinated antibody serum samples using ELISA technique indicated the significant result of optical density mean at 0.603 and 0.135, respectively. This study revealed that the production of SARS-COV-2 N recombinant protein could be carried out in E. coli expression system under denatured conditions, therefore the methods are more effective in producing the protein as a basic material in immuno-diagnostic assay

Expression of APP, CDK5, and AKT1 Gene Related to Alzheimer Disease in Brain of Long-tailed Macaques

Amyloid plaques and Neurofibrillary Tangles (NFTs) are known to be key pathological features of Alzheimer disease. To gain a better understanding of this disease, studies were carried out on the Indonesian primates, the long-tailed macaques, using a spontaneous Alzheimer's disease model. Examining and identifying genetic markers involved in plaque formation and NFTs in long-tailed macaques is necessary to reveal their physiological processes. In this study, the expression of genes involved in the development of amyloid plaque (Amyloid Precursor Protein (APP)) and those that control the phosphorylation of tau protein (CDK5 and AKT1) was examined in the long-tailed macaque brain. This study showed that APP, CDK5, and AKT1 may potentially be developed as genetic markers of Alzheimer's disease. Long-tailed macaques exhibited the development of amyloid plaque in the aging brain based on the analysis of the gene expression profile of its biomarker. Furthermore, long-tailed macaques can be optimized for neurodegenerative models

Dengue Virus Type 3 (DENV-3) Distribution in Tissues of Pig-tailed Macaque (Macaca nemestrina) Post Infection Using Immunohistochemistry Technique

Nonhuman primates play as an indispensable animal model in biomedical research for studying a variety of human health issues, diseases and disorders, therapies, and preventive strategies. In this study, we used pig-tailed macaques (Macaca nemestrina) as an experimental animal to study dengue type-3 virus (DENV-3) infection. We evaluated DENV-3 distribution and replication sites after a primary infection in all collected tissue by immunohistochemistry to localize viral protein. Significant gross lesions were not seen in any of the examined pigtailed macaques. However, microscopic lesions were present in variable degrees of severity in multiple tissues: liver, stomach, spleen and lymph nodes as evaluated by histopathological analysis. Viral protein was demonstrated in reactive lymphoid cells in spleen, thymus, axillary lymph node, inguinal lymph node, mesenteric lymph node and submandibular lymph node. In general, evidence for the presence of viral protein in various tissues after DENV-3 infection reveals that M. nemestrina is susceptible to the infection and could serve as a good alternate model to evaluate the replication of dengue virus in tissues

IDENTIFICATION AND MOLECULAR CHARACTERIZATION OF BOVINE HERPERVIRUSES (BoHV) DNA TERMINASE PARTIAL GENE IN ACEH CATTLE

Bovine Herpesvirus (BoHV) is a member of Herpesviridae family that acts as pathogenic virus causing infectious bovine rhinotracheitis (IBR) among cattles, resulting in economic loss for cattle industry. BoHV-1 infection in cows is closely related to abortion, respiratory infection, reduced milk production, infertility, and low birth weight. The aim of this study was to identify and characterize the molecular of BoHV-1 and other virus types, as well as the possible presence of other Herpesviridae family using PCR to amplify DNA terminase gene. Four out of 210 nose swab samples were positive for herpes virus on DNA terminase gene. Further characterization of samples showed 99-100% similarity to BoHv-1 and BoHV-6 sequence. Genetic distance between genera BoHV-1 and BoHV-6 is 0.518 and within genera was 0.001 and 0.044. According to phylogenetic tree analysis of DNA terminase gene, the analyzed sequence clustered into 2 genera, namely Varicellovirus which is identical to BoHV-1 and Macavirus which is identical to BoHV-6. The study provides scientific information on molecular characteristics of Herpesviridae family, especially BoHV-1 which is prevalent in Indonesia with the highest density in the central ranches in Aceh provinc

Serological Survey for Hepadnavirus in Long-tailed Macaques (Macaca fascicularis) at their ex-situ Habitats in Indonesia

The discovery of hepatitis B virus (HBV) infection among long-tailed macaques from Mauritus Island in 2013 is a new finding that showed HBV could infect non-human primate family Cercopithecidae. The aim of this study is to investigate the prevalence of Hepadnavirus among Macaca fascicularis that are living outside of their natural habitat in Indonesia. Hepatitis B surface antigen (HBsAg) screening test was performed on 95 plasma and serum samples collected from the different sources captivity, confiscated long-tailed macaques, and performance monkey. DNA detection was carried out on seropositive samples to HBsAg. Screening test showed that 11 of 95 (11.6%) samples were reactive to HBsAg. Prevalence of HBsAg is higher in confiscated animals and performance monkey (55%) compared with captive M. fascicularis (45%). However, no HBV DNA could be detected in HBsAg samples that were tested. HBsAg positive result indicate that the long-tailed macaques could be infected by hepatitis B virus naturally, although HBV DNA could not be detected in this study