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Objective: This study analyzes the mycobiome in wild and captive Sumatran orangutans. Materials and Methods: Nine orangutan feces samples from the wild and nine from captivity were divided into three repeats from 11- to 15-year-olds in good health. The Illumina platform for analysis of ITS bioinformatics was used according to the Qiime2 and CCMetagen approaches. Results: Wild Sumatran orangutans include 53% Ascomycota, 38% uncultured fungi, and 4% Basidiomycota. Orangutans in captivity are 57% Ascomycota, 26% uncultured fungi, and 2% Basidiomycota. Based on genus level, uncultured Neurospora (31%), Penicillium (10%), Aspergillus (3%), Fusarium (3%), Candida (2%), Cutaneotrichosporon (2%), and Limonomyces (2%) are found in wild orangutans. The most prevalent genus among captivity orangutans is Aspergillus (32%), followed by fungal sp. (11%), Lasiodiplodia (18%), Devriesia (2%), and Sordariomycetes (2%). According to the Chao1 diversity index and Shannon and Simpson, there was no significant difference between wild and captive Sumatran orangutans. Conclusion: Neurospora is unique to wild Sumatran orangutans, although Aspergillus predom¬inates in captive orangutans. We hypothesize that the gut mycobiome of wild orangutans will resemble that of orangutans in captivity. The excellent range of food sources in the forest does not result in the prevalence of fungi in the typical gut microbiome.

Background Indonesia is one of the countries that is endemic to avian influenza virus subtype H9N2. This study aims to compare the molecular characteristics of avian influenza virus (AIV) subtype H9N2 from West Java. Methods Specific pathogen-free (SPF) embryonated chicken eggs were used to inoculate samples. RNA extraction and RT-qPCR confirmed the presence of H9 and N2 genes in the samples. RT-PCR was employed to amplify the H9N2-positive sample. Nucleotide sequences were obtained through Sanger sequencing and analyzed using MEGA 7. Homology comparison and phylogenetic tree analysis, utilizing the neighbor-joining tree method, assessed the recent isolate's similarity to reference isolates from GenBank. Molecular docking analysis was performed on the HA1 protein of the recent isolate and the A/Layer/Indonesia/WestJava-04/2017 isolate, comparing their interactions with the sialic acids Neu5Ac2-3Gal and Neu5Ac2-6Gal. Results RT-qPCR confirmed the isolate samples as AIV subtype H9N2. The recent virus exhibited 11 amino acid residue differences compared to the A/Layer/Indonesia/WestJava-04/2017 isolate. Phylogenetically, the recent virus remains within the h9.4.2.5 subclade. Notably, at antigenic site II, the recent isolate featured an amino acid N at position 183, unlike A/Layer/Indonesia/WestJava-04/2017. Molecular docking analysis revealed a preference of HA1 from the 2017 virus for Neu5Ac2-3Gal, while the 2023 virus displayed a tendency to predominantly bind with Neu5Ac2-6Gal. Conclusion In summary, the recent isolate displayed multiple mutations and a strong affinity for Neu5Ac2-6Gal, commonly found in mammals.

Background and Aim: Klebsiella pneumoniae is one of the respiratory disease agents in human and chicken. This bacterium is treated by antibiotic, but this treatment may trigger antibiotic resistance. Resistance gene in K. pneumoniae may be transferred to other bacteria. One of the known resistance genes is extended-spectrum β-lactamase (ESBL). This research aimed to study K. pneumoniae isolated from chicken farms in East Java, Indonesia, by observing the antibiotic resistance pattern and detect the presence of ESBL coding gene within the isolates. Materials and Methods: A total of 11 K. pneumoniae isolates were collected from 141 chicken cloacal swabs from two regencies in East Java. All isolates were identified using the polymerase chain reaction method. Antimicrobial susceptibility was determined by agar dilution method on identified isolates, which then processed for molecular characterization to detect ESBL coding gene within the K. pneumoniae isolates found. Results: The result of antibiotic sensitivity test in 11 isolates showed highest antibiotic resistance level toward ampicillin, amoxicillin, and oxytetracycline (100%, 100%, and 90.9%) and still sensitive to gentamicin. Resistance against colistin, doxycycline, ciprofloxacin, and enrofloxacin is varied by 90.9%, 54.5%, 27.3%, and 18.2%, respectively. All isolates of K. pneumoniae were classified as multidrug resistance (MDR) bacteria. Resistance gene analysis revealed the isolates harbored as blaSHV (9.1%), blaTEM (100%), and blaCTX-M (90.9%). Conclusion: All the bacterial isolates were classified as MDR bacteria and harbored two of the transmissible ESBL genes. The presence of antibiotic resistance genes in bacteria has the potential to spread its resistance properties.

This study aimed to isolate and identify from broiler samples from Sukabumi, Indonesia. Also, antibiogram studies of the isolated bacteria were carried out considering the detection of the antibiotic resistance genes. Cloaca swabs ( = 45) were collected from broilers in Sukabumi, Indonesia. Isolation and identification of were carried out according to standard bacteriological techniques and biochemical tests, followed by confirmation of the polymerase chain reaction targeting the gene. Antibiotic sensitivity test, using several antibiotics [tetracycline (TE), oxytetracycline (OT), ampicillin (AMP), gentamicin (CN), nalidixic acid (NA), ciprofloxacin (CIP), enrofloxacin (ENR), chloramphenicol, and erythromycin] was carried out following the Kirby-Bauer disk diffusion method. Detection of antibiotic resistance coding genes was carried out by PCR using specific oligonucleotide primers. Statistical analysis was carried out with one-way analysis of variance. The results showed that 55.6% (25/45) of the samples were associated with the presence of . Antibiotic sensitivity test showed that the isolates were resistant to TE (88%; 22/25), OT (88%; 22/25), AMP (100%; 25/25), CN (64%; 16/25), NA (100%; 22/25), CIP (88%; 22/25), ENR (72%; 18/25), chloramphenicol (0%; 0/25), and erythromycin (92%; 23/25). On the other hand, the antibiotic resistance coding genes were (86.4%; 19/22), (100%; 25/25), (0%; 0/16), (100%; 25/25), and (13%; 3/23). It was found that chloramphenicol is markedly different from other antibiotic treatment groups. was successfully isolated from cloacal swabs of broiler in Sukabumi, Indonesia. The bacteria were resistant to TE, OT, AMP, CN, NA, CIP, ENR, and erythromycin. Chloramphenicol was more sensitive and effective than other antibiotics in inhibiting the growth of The antibiotic resistance genes detected were , , , and .

Objective: Orangutans (Pongo abelii), as endemic primates of Indonesia, are characterized by a predominantly arboreal lifestyle. Klebsiella pneumoniae (K. pneumonia) and other Gram-negative bacteria are present in the Indigenous flora of many mammals, including orangutans. This study aimed to investigate the antibiotic resistance and virulence profile of K. pneumonia isolated from wild Sumatran orangutans. Materials and Methods: This study investigated 10 fecal samples from wild Sumatran orangutans from the Gunung Leuser National Park, Aceh, Indonesia. Biochemical and molecular identification of K. pneumoniae using the RNA polymerase subunit b gene and detection of virulence-associated genes. In addition, molecular detection of antibiotic resistance genes was performed to character¬ize the resistance mechanisms in the isolates. Results: K. pneumonia was detected in 6 out of 10 fecal samples from wild Sumatran orangutans. The virulence genes mrkD and entB were detected in all (100%) of the isolates, whereas wabG was identified in 83.33% of the strains. Antibiotic susceptibility testing against K. pneumoniae revealed that three isolates were susceptible to streptomycin (S) and nalidixic acid (NA), while all six isolates were susceptible to chloramphenicol and ciprofloxacin. One isolate demonstrated intermediate resistance to NA, while the remaining two exhibited intermediate resistance to S. Six isolates were resistant to ampicillin, tetracycline, and erythromycin, indicating multidrug resis¬tance. Furthermore, antibiotic resistance genes were detected in the isolates with the following prevalence: blaTEM gene (six isolates; 100%), blaSHV (six isolates; 100%), blaCTX-M gene (four isolates; 66.67%), and tetA gene (four isolates; 66.67%). Conclusion: This study revealed the virulence and resistance profile of K. pneumoniae bacterium isolated from wild Sumatran orangutans, which is essential for formulating effective conservation and healthcare strategies.

The incidence of salmonellosis in humans and animals is still high due to the occurrence of virulence factors in which play a role in the process of infection in the host and the spread of disease and most of the can infect humans and animals. The present study was aimed to identify and detect virulence genes related to pathogenicity islands (SPIs) and plasmid virulence (Spv). A total of 27 . archive isolates belonging to the National Veterinary Drug Assay Laboratory (NVDAL) were used in this study. The bacteria were collected in 2016 and 2017 from samples of the cloaca and fecal swabs from layer and broiler farms in five provinces of Java Island. Isolates were cultured in specific media, biochemical tests and Gram staining. Detection of . and virulence genes was done by polymerase chain reaction (PCR) method. Identification of serovar showed 100% (27/27) isolates were positive for the gene (304 bp). The result confirmed that all strains were . PCR based detection of virulence genes showed that 100% of isolates had virulence genes in SPI-1 to SPI-5, namely, and genes. All the isolates (27/27) were also positive to gene-based PCR. All the isolates of in this study carry virulence genes related to SPI-1 to SPI-5 and plasmid virulence. The existence of virulent genes indicates that the strain examined in this study is highly virulent and poses a potential threat of worse disease outcome in humans and animals.

Background Indonesia is one of the countries that is endemic to avian influenza virus subtype H9N2. This study aims to compare the molecular characteristics of avian influenza virus (AIV) subtype H9N2 from West Java. Methods Specific pathogen-free (SPF) embryonated chicken eggs were used to inoculate samples. RNA extraction and RT-qPCR confirmed the presence of H9 and N2 genes in the samples. RT-PCR was employed to amplify the H9N2-positive sample. Nucleotide sequences were obtained through Sanger sequencing and analyzed using MEGA 7. Homology comparison and phylogenetic tree analysis, utilizing the neighbor-joining tree method, assessed the recent isolate's similarity to reference isolates from GenBank. Molecular docking analysis was performed on the HA1 protein of the recent isolate and the A/Layer/Indonesia/WestJava-04/2017 isolate, comparing their interactions with the sialic acids Neu5Ac2-3Gal and Neu5Ac2-6Gal. Results RT-qPCR confirmed the isolate samples as AIV subtype H9N2. The recent virus exhibited 11 amino acid residue differences compared to the A/Layer/Indonesia/WestJava-04/2017 isolate. Phylogenetically, the recent virus remains within the h9.4.2.5 subclade. Notably, at antigenic site II, the recent isolate featured an amino acid N at position 183, unlike A/Layer/Indonesia/WestJava-04/2017. Molecular docking analysis revealed a preference of HA1 from the 2017 virus for Neu5Ac2-3Gal, while the 2023 virus displayed a tendency to predominantly bind with Neu5Ac2-6Gal. Conclusion In summary, the recent isolate displayed multiple mutations and a strong affinity for Neu5Ac2-6Gal, commonly found in mammals.

Background and Aim: Pathogenic Escherichia coli contamination along the broiler meat supply chain is a serious public health concern. This bacterial infection with multidrug-resistant can lead to treatment failure. Several studies have revealed that avian pathogenic E. coli (APEC) and human extraintestinal pathogenic E. coli (ExPEC) showed a close genetic relationship and may share virulence genes. This study aimed to determine the phylogenetic group and virulence gene profiles in colistin-resistant E. coli obtained from the broiler meat supply chain in Bogor, West Java, Indonesia. Materials and Methods: Fifty-eight archive isolates originated from the cloacal swab, litter, drinking water, inside plucker swab, fresh meat at small scale poultry slaughterhouses, and traditional markets were used in this study. All the isolates were characterized by a polymerase chain reaction to determine the phylogenetic group (A, B1, B2, or D) and virulence gene profiles with APEC marker genes (iutA, hlyF, iss, iroN, and ompT). Results: Phylogenetic grouping revealed that the isolates belong to A group (34.48%), D group (34.48%), B1 group (17.24%), and B2 group (13.79%). The virulence gene prevalence was as follows: iutA (36%), hlyF (21%), ompT (21%), iroN (10%), and iss (9%). The B2 group presented with more virulence genes combinations. iroN, hlyF, and ompT genes were positively associated with the B2 group (p≤0.05). Conclusion: Our results highlight the role of colistin-resistant E. coli originated from the broiler meat supply chain as a potential reservoir for human ExPEC virulence genes.

This study aimed to identify genes encoding resistance to tetracycline (TE) and plasmid-mediated resistance to quinolones in isolates from clinical cases of avian colibacillosis in Sukabumi, Indonesia. A total of 25 archive isolates were collected in 2013-2017 from clinical cases of avian colibacillosis in Sukabumi, Indonesia. All isolates were tested for TE and quinolone resistance using the disk diffusion method. TE -resistant isolates were screened for the presence of and genes by single polymerase chain reaction (PCR). The ( ), ( ), and ( ) genes were detected by multiplex PCR in quinolone-resistant isolates. Result of this study shows that 19 of 25 (76%) isolates are resistant to oxytetracycline and 64% are resistant to TE; among them, 63.2% and 31.5% were positive and , respectively. 13 out of 25 (52%) are resistant to ciprofloxacin and 36% are resistant to enrofloxacin either norfloxacin; among them, 61.6% were positive ( ), 7.7% were positive ( ), 23% were positive ( ), and 7.7% were positive both of ( ) and ( ). This study shows that a few pathogens of are resistant to TE and quinolone. The frequency of and genes that are responsible for this resistance among avian pathogenic isolates in Sukabumi, Indonesia, was high.

Objective: This study aims to evaluate the effect of Clostridium perfringens sialidase treatment on monolayer cell behavior using computational screening and an in vitro approach to demonstrate interaction between enzyme-based drugs and ligands in host cells. Materials and Methods: The in silico study was carried out by molecular docking analysis used to predict the interactions between atoms that occur, followed by genetic characterization of sialidase from a wild isolate. Sialidase, which has undergone further production and purification processes exposed to chicken embryonic fibroblast cell culture, and observations-based structural morphology of cells compared between treated cells and normal cells without treatment. Results: Based on an in silico study, C. perfringens sialidase has an excellent binding affinity with Neu5Acα (2.3) Gal ligand receptor with Gibbs energy value (ΔG)—7.35 kcal/mol and Ki value of 4.11 μM. Wild C. perfringens isolates in this study have 99.1%–100% similarity to the plc gene, NanH, and NanI genes, while NanJ shows 93.18% similarity compared to the reference isolate from GenBank. Sialidase at 750 and 150 mU may impact the viability, cell count, and cell behavior structure of fibroblast cells by significantly increasing the empty area and perimeter of chicken embryo fibroblast (CEF) cells, while at 30 mU sialidase shows no significant difference compared with mock control. Conclusion: Sialidase-derived C. perfringens has the capacity to compete with viral molecules for attachment to host sialic acid based on in silico analysis. However, sialidase treatment has an impact on monolayer cell fibroblasts given exposure to high doses.