Dairy farmers' knowledge, attitudes, and practices regarding the brucellosis surveillance and control program in Bogor, Indonesia.

Background and Aim: Brucellosis is an infectious and zoonotic disease that affects people's health and the economy in most countries. Brucellosis is still prevalent in several Indonesian regions. This study aimed to analyze the correlation between the characteristics, knowledge, attitudes, and practices (KAP) of dairy farmers in Bogor District in supporting brucellosis control and surveillance programs. Materials and Methods: The study was cross-sectional. Data were collected through interviews with 151 dairy farmers in Bogor Regency, West Java, Indonesia. The outcome is brucellosis surveillance and control practice among dairy farmers, and the variables include individual characteristics, knowledge, and attitudes toward brucellosis surveillance and control. Descriptive analysis and path analysis were used in statistical analysis. Results: The majority of farmers' knowledge, attitudes and practices were moderate, with the percentages 67.55%, 60.92%, and 41.72% respectively. Formal education, training, and dairy rising length are variables that have a direct and significant impact on knowledge level. Knowledge is the variable that influences the overall level of attitude. Age, knowledge, and attitude are factors that influence the practice of brucellosis surveillance and control. Conclusion: Although the practice level of brucellosis surveillance and control for dairy farmers in Bogor Regency is moderate, efforts to improve it are still required. The basic effort is critical for increasing farmers' knowledge.

A preliminary metagenomics study of bacteria present in the dirt of Swiftlet farmhouses based on nitrite levels in edible bird's nest on Sumatera Island, Indonesia.

Background and Aim: Since the past decade, metagenomics has been used to evaluate sequenced deoxyribonucleic acid of all microorganisms in several types of research. Nitrite contamination originates from the natural environment in Swiftlet farmhouses (SFHs) and can influence nitrite levels in edible bird's nest (EBN). It is strongly speculated that the conversion process into nitrite is influenced by the bacteria present in SFHs. Nitrite can cause adverse effects on human health. The previous research has focused on the characteristics of bacteria that may influence the nitrite conversion process in SFHs. This study aimed to a metagenomics analysis of bacteria present in the dirt of SFHs and evaluated nitrite levels in EBN on Sumatera Island. Materials and Methods: In total, 18 SFHs on Sumatera Island were selected, and EBN and dirt samples were collected from each SFH, resulting in 18 EBN and 18 dirt SFH samples. Raw uncleaned white EBN and dirt from three areas of SFH were collected. The samples were analyzed for nitrite levels using a spectrophotometer, and the metagenomics sequencing of SFH dirt samples was performed using the MinIon nanopore method. The sequenced data were analyzed using the EPI2ME software. Results: Of the 18 raw uncleaned white EBN samples, 9 (50%) had <30 ppm nitrite levels. The top five bacterial genera in SFH dirt samples in Group A (nitrite levels >30 ppm) were Aeromonas, Escherichia, Acinetobacter, Arcobacter, and Acetoanaerobium. Those in Group B (nitrite levels <30 ppm) were Aeromonas, Pseudomonas, Shewanella, Escherichia, and Acinetobacter. There were 12 genera of nitrifying bacteria in Group A and 8 in Group B. The total cumulative read of nitrifying bacteria in Groups A and B were 87 and 38 reads, respectively. Conclusion: This is the first study to show that characteristic bacteria present in the dirt of SFHs might significantly influence the conversion from nitrogen to nitrite. Approximately 50% of raw uncleaned EBN samples had <30 ppm nitrite levels. Aeromonas was the most dominant bacterial genus found in Groups A and B. The variations in genus and cumulative reads nitrifying bacteria in group A were greater than those in Group B. This study provides information on the characteristics of bacteria that may influence the nitrite conversion process in SFHs. Metagenomics data were obtained from the reading using the software EPI2ME. Further research is needed on the bacterial target species that can convert nitrite in SFHs.

A sequential toggle cell-SELEX DNA aptamer for targeting Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli bacteria.

BACKGROUND: Mastitis is an inflammation of the mammary glands caused by a microbial infection. The common bacteria causing this infection in dairy farms are Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. The aptamer is a new biosensor platform for detecting pathogens; however, its use for simultaneous detection of S. aureus, S. agalactiae, and E. coli bacteria has not been reported. This study's objective is to isolate and characterize polyclonal DNA aptamer with broad reactivity to the mastitis bacteria S. aureus, S. agalactiae, and E. coli using a sequential toggle cell-SELEX. METHODS AND RESULTS: The DNA aptamer pool from SELEX 15 was inserted into the pGEM-T easy plasmid. Furthermore, the transformant clones were selected by PCR colony, plasmid isolation, and sequencing. Six DNA aptamers, consisting of S15K3, S15K4, S15K6, S15K13, S15K15, and S15K20 with a constant region and the right size of 81 bp were derived from the sequencing analysis. The secondary structure of the DNA was predicted using Mfold software. The DNA was analyzed with binding characteristics, including binding capacity and affinity (Kd), using qPCR. The results indicated aptamer S15K15 has the highest binding ability into S. agalactiae, while S15K13 performed binding capacity most to E. coli EPEC 4, and S15K3 has the highest capacity of binding to S. aureus BPA-12. CONCLUSION: Aptamer S15K3 has the best binding characteristics on all three bacterial targets.

An investigation of heavy metals in edible bird's nest from Indonesia using inductively coupled plasma mass spectrometry.

Background and Aim: In 2020, Indonesia, which has the highest global production of edible bird's nest (EBNs), exported up to 1312.5 tons of this product at a value of USD 540.4 million. Recently, food safety aspects related to EBNs, including contamination with heavy metals, have become a serious concern. However, data on the presence and concentration of heavy metals in EBNs in Indonesia are not yet available. This study aimed to determine and compare the presence and concentrations of arsenic (As), mercury (Hg), lead (Pb), cadmium (Cd), and tin (Sn) in EBNs originating from several primary Indonesian islands. The study also analyzed the effect of washing on the heavy metal content in EBNs. Materials and Methods: A study on 44 swiftlet farmhouses (SFHs) was conducted to determine the concentrations of heavy metals in EBNs. The number of samples from the SFHs was allocated proportionally to the main EBN-producing islands in Indonesia, that is, Kalimantan, Sumatera, Sulawesi, and Java (22, 13, 7, and 2, respectively). The concentrations of the above five elements in the samples before washing (raw-unclean EBNs) and after washing (raw-clean EBNs) were determined by inductively coupled plasma mass spectrometry. Washing was conducted according to the general procedures at an EBN processing plant. Results: The raw-unclean EBNs from the four islands contained As, Pb, Cd, and Sn at varying concentrations. However, Hg was not detected in the raw-unclean EBN samples from Sulawesi. The raw-unclean EBNs from Kalimantan had lower concentrations of Pb and Cd compared with the other islands. The concentrations of As, Pb, Cd, and Sn in the EBNs decreased significantly after washing with clean water. Conclusion: Heavy metals (As, Hg, Pb, Cd, and Sn) were detected at a low level in most of the raw-unclean EBNs originating from the main Indonesian island where they were produced. The concentrations of all the heavy metals reviewed in the raw-unclean EBNs samples decreased significantly after washing.

Chemical compositions, contaminants, and residues of organic and conventional goat milk in Bogor District, Indonesia.

AIM: This study aimed to compare chemical composition and contaminants (pesticide residues, antibiotic residues, and heavy metal residues) between organic and conventional goat milk in Bogor District, West Java Province, Indonesia. MATERIALS AND METHODS: Milk sampling was carried out from March to August 2018 at six goat farms. The chemical quality of milk was checked using the Lactoscan Ultrasonic Milk Analyzer device. Fatty acids were analyzed using gas chromatography (GC). Pesticide residues in goat's milk were analyzed using a GC-electron capture detector (GC-ECD). Antibiotic residues were analyzed using bioassay screening test method. The lead (Pb) and arsenic (As) residues were analyzed using the Atomic Absorption Spectrophotometer (AAS). RESULTS: The content of fat, protein, and lactose showed that there was no difference in the composition of goat's milk between organic and conventional farms. Caprylic acid (C8:0) and capric acid (C10:0) of organic goat milk are higher than conventional goat milk. Stearic acid (C18:0) and linoleic acid (C18:2) of conventional goat milk are higher than organic goat milk. The total fatty acid of organic goat milk is higher than conventional goat milk. Organochlorine pesticide residues were not detected in organic goat milk and conventional goat milk. Tetracycline antibiotic residues were found in one sample (5.56%) of organic goat milk, and macrolides residues were found in two samples (11.11%) of conventional goat milk. Pb residue in organic goat milk is 50 ppb while conventional goat milk is 80 ppb. Residue As in organic goat milk is 70 ppb while conventional goat milk is 110 ppb. CONCLUSION: There was no chemical composition (fat, protein, and lactose) difference between organic and conventional goat milk. Saturated fatty acid (SFA) in organic goat milk is higher than conventional goat milk. Pesticide residues are not found in both organic and conventional goat milk. Tetracycline antibiotics were found in organic goat milk and macrolide antibiotic groups found in conventional goat milk. Pb and As residues were found in both organic goat milk and conventional goat milk.

Diversity of lactic acid bacteria in dadih produced by either back-slopping or spontaneous fermentation from two different regions of West Sumatra, Indonesia.

AIM: Dadih samples from two different origins (Kamang and Gadut in West Sumatra) manufactured with different methods (back-slopping or spontaneous fermentation) were evaluated for the diversity of lactic acid bacteria (LAB). MATERIALS AND METHODS: Four dadih samples manufactured with two different fermentation methods were obtained from Kamang and Gadut regions. Both genotypic and phenotypic characteristic (16S rRNA partial gene sequence analysis and carbohydrate fermentation profile) were used to analyze the diversity of dadih LAB population. RESULTS: This study showed that LAB count in back-slopping fermented dadih was one log cycle higher than spontaneous fermented dadih. LAB isolates from the two regions were divided into three genera, namely Lactococcus, Lactobacillus, and Pediococcus. Sequencing results showed that 41.6% (five isolates) were identified as Lactococcus lactis ssp. lactis, 25% (three isolates) were identified as Lactobacillus plantarum ssp. plantarum, 16.6% (two isolates) were identified as L. lactis ssp. cremoris, and 8.3% (one isolate each) were identified as Pediococcus pentosaceus and Lactobacillus pentosus. CONCLUSION: Five species were determined in back-slopping fermented dadih, i.e., L. lactis ssp. lactis, L. lactis ssp. cremoris, L. plantarum ssp. plantarum, L. pentosus, and P. pentosaceus. On the other hand, spontaneous fermented dadih only contained three different species, namely L. lactis ssp. lactis, L. lactis ssp. cremoris, and L. plantarum ssp. plantarum. This research showed that back-slopping fermentation offers greater abundance and diversity compared to spontaneous fermentation in dadih.

Detection of multi-drug resistant (MDR) Escherichia coli and tet gene prevalence at a pig farm in Kupang , Indonesia.

OBJECTIVE: The purpose of this study was to detect the incidence of multi-drug resistant (MDR) and the spread of tet genes that encode tetracycline (TE) resistance in E. coli in pig farms in the city of Kupang, Indonesia. MATERIALS AND METHODS: Samples of pig feces have been obtained from 96 pig farms in Kupang city, Indonesia. Escherichia coli bacteria were isolated and identified morphologically and biochemically, and finally confirmed by the API test. The disk diffusion method has been used to observe the antibiotic sensitivity effects and has been followed by observing resistant genes encoding TE resistance using the multiplex polymerase chain reaction (m-PCR) method to detect the presence of tet genes such as tet (A), tet (B), tet (C), tet (D), and tet (E), respectively. RESULTS: A total of 82 (85.4%) of E. coli isolates have been found in all pig feces samples obtained from 96 pig farms in Kupang city. This study has shown a high level of antibiotic resistance dominated by erythromycin (85.4%) and cephalothin (58.5%) and followed by several other antibiotics with a percentage below 34.1%. The prevalence of MDR E. coli was 57.3% by showing 39 different patterns. The most common pattern was showed by the Cephalothin-Colistin-Erythromycin pattern. The resistance of E. coli to TE appears to be related to the presence of tet (A) and tet (E) genes. CONCLUSION: This study has encouraged the need for public awareness (farmers) of the wise use of antibiotics in preventing the spread of resistant bacteria that can cause health problems in animals and humans.

Extended-Spectrum ß-Lactamase (ESBL)-Producing Klebsiella pneumoniae in Bulk Tank Milk from Dairy Farms in Indonesia.

Bulk tank milk from 80 dairy farms located in the West Java Region of Indonesia was analyzed for the presence of extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae. Isolates from seven dairy farms were ESBL positive, and all were identified as Klebsiella pneumoniae. The isolates showed ESBL-characteristic antibiotic resistance patterns. Further analysis revealed that all K. pneumoniae isolates harbored the blaSHV gene, and two isolates were additionally positive for the blaTEM-1 and blaCTX-M-15 genes. Isolates from different farms were clonally diverse according to macrorestriction analysis. The results indicate that the relatively high frequency of ESBL-producing K. pneumoniae in bulk tank milk implies the risk that milk is both a source of local exposure and a vector contributing to the supraregional spread of antibiotic-resistant bacteria by trade.