HAYATI Journal of Biosciences

Immune Response to Capsular Polysaccharide of Streptococcus pneumoniae in Rabbits Immunized with Pneumococcal Conjugate Vaccine

Fri, 24 Jan 2025 13:46:17 +0700

To evaluate the immune response of a rabbit model to polysaccharide capsules of Streptococcus pneumoniae after primary and booster immunization with pneumococcal conjugate vaccine. Rabbits were immunized with 0.25 ml of the 13-valent pneumococcal conjugate vaccine (PCV13) on Day 0, 7, and 14. Blood of rabbits was collected to measure the specific antibodies against the native polysaccharide capsules of S. pneumoniae serotypes 6B (Anti-Pn6BPS antibodies) and 19F (Anti-19FPS antibodies) using enzyme-linked immunosorbent assay (ELISA). The specific anti-Pn6BPS and anti-Pn19FPS antibodies in the immunized group increased significantly compared to the control rabbit after immunization (P<0.05). The highest titer of anti-Pn6BPS and anti-Pn19FPS antibodies were obtained on Day 21 and Day 28, respectively. The third immunization on Day 21 is a fairly high increase in specific antibodies against polysaccharide type 6B and type 19F. The highest antibody titers against polysaccharide capsules of S. pneumoniae were obtained after booster immunization.

Impact of Dietary Variation on Gut Microbiota Composition in Macaca fascicularis at the Primate Research Center, IPB University

Anggitha Tambunan, Puji Rianti, Huda Shalahudin Darusman, Randall C. Kyes — Mon, 03 Feb 2025 10:17:07 +0700

Long-tailed macaques (Macaca fascicularis: LTMs) are widely distributed, adaptable primates commonly used as animal models in biomedical research. At primate facilities such as the Dramaga Captive Breeding Facility and the Tinjil Island Natural Habitat Breeding Facility at the Primate Research Center, IPB University, LTMs experience varying diets depending on their captive or semi-wild environments. At Dramaga Captive Breeding Facility, LTMs housed in individual and group cages are provided with different feeding regimens, whereas the semi-wild LTMs on Tinjil Island feed primarily on natural food sources with occasional, very modest provisioning of bananas. Our study's objective is to evaluate these diets' influence on the gut microbiota and overall health of LTMs. Through comparative analysis of fecal microbiomes using 16S rRNA sequencing, we identified distinct gut microbiota profiles between captive and semi-wild LTMs. Semi-wild LTMs displayed a predominance of Proteobacteria, notably Succinivibrio, associated with a high-fiber diet derived from wild plant consumption. In contrast, captive LTMs housed in individual and group cages exhibited dominance of Firmicutes and Bacteroidota, respectively. These findings suggest that diet is pivotal in shaping gut bacterial profiles, potentially impacting metabolic functions. Our study provides insight into the relationship between certain diets and gut bacterial composition in LTMs across different breeding facilities at the Primate Research Center, IPB University, thereby offering valuable implications for LTMs' welfare and management strategies.

Antibacterial, Antifungal, Antioxidant, and Photoprotective Analysis of Mangrove Extracts as Additives Ingredients in a Cosmetic Cream

Dear Berliana Putri, Delianis Pringgenies, Agus Trianto — Fri, 10 Jan 2025 15:32:42 +0700

Mangroves are a rich source of natural antioxidant compounds that can inhibit and reduce tissue damage caused by free radical activity. Currently, cosmetic creams are widely recommended for maintaining skin health and aesthetics, especially in protecting against the harmful effects of free radicals. This study applied mangrove extract as an additive ingredient in a cosmetic cream, serving as an antibacterial, antifungal, antioxidant, and photoprotective agent. Mangrove samples were collected from the mangrove ecosystem in Tapak Village, Semarang, Indonesia. Three mangrove species were evaluated for their bioactivity, and the species with the highest bioactivity was selected for use in the cream formulation. Antibacterial and antifungal activities were tested using the disc diffusion method, antioxidant activity was assessed using the DPPH method, and photoprotective activity was determined using UV spectrophotometry, with analysis based on the Mansur mathematical equation. The cream was evaluated for its characteristics, including spreadability, homogeneity, sensory properties, pH, sun protection factor (SPF), phytochemical content, stability, and microbial contamination. An in vivo was conducted to assess the cream’s effectiveness on white mice. Results indicated that Avicennia marina leaf extract exhibited the highest bioactivity compared to the other two species, Rhizophora mucronata and Bruguiera gymnorrhiza. Consequently, A. marina leaf extract was selected as the active additive ingredient in the cream formulation. Characterization tests demonstrated that the cream was stable and met standard criteria for quality. The in vivo analysis revealed that the mangrove leaf extract cream significantly prevented epidermal thinning, reduced neutrophil counts, and preserved fibroblast numbers.

Influence of Co-feeding Methanol-sorbitol Ratio on Production of Human Insulin Precursor Expressed by Mut+ Pichia pastoris

Fri, 24 Jan 2025 09:13:42 +0700

An increasing number of diabetic patients and the demand for insulin encourage the development of recombinant insulin production on a large scale. Human insulin precursor (HIP) expressed by Mut⁺ Pichia pastoris using methanol as an inducer was developed. However, methanol above 5% (w/v) or 1.56 M is toxic for the host. Sorbitol was introduced as a co-substrate with methanol. To our knowledge, the study of methanol/sorbitol co-feeding on human insulin precursor (HIP) expression by Mut⁺ Pichia pastoris in a bioreactor has yet to be reported. This study aimed to investigate the influence of the methanol-sorbitol co-feeding ratio on the expression of HIP expressed by P. pastoris X33 Mut⁺. The study was conducted by comparing the cultivation of P. pastoris/pD902-IP Mut⁺ in a 10-liter bioreactor under three conditions: feeding 100% methanol, mass ratio of MeOH:sorbitol 12:1 and 3:1. The oxygen consumption of methanol/sorbitol is less than the methanol feeding. The mass ratio of MeOH:sorbitol 12:1 produced the highest HIP titer (1326.5 mg/L), 1.5 times higher than methanol feeding, the lowest specific growth rate, but the highest specific productivity at the induction phase. MeOH:sorbitol mass ratio 3:1 produced the highest dry cell weight (DCW) amount (96 g/L). These results suggested that an appropriate ratio of sorbitol-methanol can be a choice to replace methanol feeding in a Mut⁺ P. pastoris.

Congenital Malformations in Chicken Embryos After Oxybenzone Exposure

Fri, 24 Jan 2025 09:31:20 +0700

Topical use of oxybenzone, commonly found in sunscreens, can be absorbed by the skin, and long-term use may cause endocrine disruption, cancer, and teratogenic effects. However, its potential teratogenic effects on embryonic development have not been well-studied. This study aims to determine the impact of oxybenzone exposure on the early stage of embryonic development. Chicken embryos aged 72 hours (20 Hamburger-Hamilton/HH stage) were exposed to a pure oxybenzone for 24 hours at varying concentrations (0, 1, 5, 10, and 20 ppm), each group consisting of 3 embryos. Embryo preparations were made using the wholemount method. Morphological abnormalities were observed with a stereo microscope, and descriptively morphometric measurements were analyzed using ImageJ software. Statistical analysis used One-way ANOVA and Tukey’s test for normally distributed data, while Kruskal-Wallis H and Mann-Whitney U test for non-normally distributed data. This study found that oxybenzone significantly enlarged the embryo, telencephalon, and eye. Several abnormalities were observed in the embryos exposed to oxybenzone, including incomplete closure of the anterior neuropore, concavity in the anterior and lateral of the mesencephalon, and depressions in the tail bud. This study concludes that oxybenzone acts as a teratogen, causing abnormalities in embryonic development, particularly in the central nervous system.

Screening and Molecular Identification of Endophytic Bacteria from Calamus caesius Blume with Potential as Antioxidant and α-Glucosidase Inhibitors

Chrismis Novalinda Ginting, Nyoman Ehrich Lister, Ermi Girsang, Edy Fachrial — Fri, 24 Jan 2025 09:45:12 +0700

Calamus caesius, traditionally used in Asian medicine for managing diabetes mellitus, is a promising source of endophytic bacteria due to its bioactive potential. This study aims to identify the potential of endophytic bacteria isolated from “Pakkat” (Calamus caesius Blume), a traditional food of North Sumatra, as a source of α-glucosidase inhibitors. These inhibitors, such as acarbose, are used to manage type 2 diabetes mellitus but have significant gastrointestinal side effects. This study focuses on isolating and characterizing endophytic bacteria that can produce natural inhibitors, potentially safer alternatives. Twenty endophytic bacterial isolates were successfully isolated, of which five were excluded due to hemolytic activity. Antioxidant activity was determined using the DPPH method, and isolates with the highest activity proceeded to α-glucosidase inhibitor testing. Molecular identification of potential isolates was performed through 16S rRNA gene amplification, and genomic DNA sequencing utilized the Oxford Nanopore Technologies platform. Secondary metabolite analysis was performed using antiSMASH, and gene cluster comparison with CAGECAT. Of 15 isolates, LTUP1.3 (Bacillus cereus) exhibited 86.6% α-glucosidase inhibitor activity, close to the positive control (88%). Although in Bacillus cereus LTUP1.3 no secondary metabolites were found that act as α-glucosidase inhibitors, three enzymes have similarities with enzymes found in Actinoplanes that play a role in acarbose biosynthesis namely dTDP-glucose 4,6-dehydratase RffG, glucose-1-phosphate thymidylyltransferase rmIA and glucose-1-phosphate adenylyltransferase glgC. These results suggest that Bacillus cereus strain LTUP1.3 has a potential biosynthetic pathway similar to the acarbose biosynthetic pathway in Actinoplanes. However, further confirmation is required to establish its overall acarbose production capability.

Characteristics of Nitrogen-Fixing Bacteria Isolated from the Rhizosphere at Kaolin Lake, Belitung, Indonesia

Nurhakiki, Iman Rusmana, Rika Indri Astuti — Fri, 24 Jan 2025 09:58:56 +0700

Former kaolin mining has poor physical and chemical properties for plants. The low nutrient content makes difficult for plants to grow. Nitrogen (N₂) is an important macronutrient for plants, but it can only be absorbed in the form of ammonium ions (NH⁴⁺) or nitrate ions (NO^3-). Therefore, it is necessary to transform nitrogen, which is able to change the molecular form so that it can be absorbed by plants. The strategy that can be done is using nitrogen-fixing bacteria. This study aimed to explore and analyze the diversity of nitrogen-fixing bacteria based on 16S rRNA and nifH gene from the rhizosphere of the Kaolin Lake area. The method started with bacterial isolation from rhizosphere soil samples. Selected isolates were tested for ammonium content to determine the ability of isolates to fix nitrogen. In this study, 13 isolates had the ability to fix nitrogen, two of which had a high ammonium concentration, namely RBN 5.6, and RBA 2.3. The highest ammonium concentration was produced by isolate RBN5.6 amounting to 7.55 μg/ml. Based on 16S rRNA gene identification, isolate RBN5.6 was similar to Burkholderia cepacia and isolate RBA2.3 was similar to Bacillus aquimaris. Based on the detection of the nifH gene, isolate RBN5.6 was identified as a protein-encoding dinitrogenase reductase gene group cluster from the Stutzerimonas stutzeri.

The Antiaging Potential of Serum Formulations from Centella asiatica, Curcuma longa, Aloe vera, Rosa centifolia, and Salmon DNA on Injured Human Fibroblast Cells

Fri, 24 Jan 2025 10:50:41 +0700

External aging factors such as UV exposure, pollution, and lifestyle choices contribute to skin aging, resulting in deep wrinkles, fine lines, and rough skin, which can lower self-confidence. Plant extracts have been widely studied for their antiaging potential, while Salmon DNA has shown promise in stimulating collagen production. This study explores the formulation of a serum combining Centella asiatica, Curcuma longa, Aloe vera, Rosa centifolia, and Salmon DNA for its antiaging effects on injured human fibroblast cells. The serum was formulated using extracts from C. asiatica, C. longa, A. vera, R. centifolia, and Salmon DNA. Antioxidant activity was evaluated with the DPPH method, cytotoxicity using the WST-8 assay, and gene expression through qRT-PCR for COL1A1, TGF-β1, HYAL-1, and FGF-2. The serum exhibited weak antioxidant activity (IC₅₀ = 373.33 µg/ml) and reduced cell viability at high concentrations. Gene expression analysis revealed increased expression of COL1A1, TGF-β1, and FGF-2, along with reduced HYAL-1 expression in injured BJ cells. The formulated serum shows potential as an antiaging agent, promoting collagen production and reducing hyaluronidase activity.

Chemical Components of Essential Oils from Woods of Cinnamomum burmanii (Ness) BL, Santalum album Linn., and Cryptocarya massoia (Oken) Kosterm and Their Application as Antibacterial Additives in Transparent Herbal Soaps

Sri Retno Dwi Ariani, Tatik Lestari — Fri, 24 Jan 2025 11:51:18 +0700

This study aimed to isolate and identify the chemical components of essential oils from woods of cinnamon (Cinnamomum burmanii (Ness) BL), sandalwood (Santalum album Linn.), and massoia (Cryptocarya massoia (Oken) Kosterm) and to determine their effect as additives for transparent herbal soap in terms of their antibacterial activity against Staphylococcus aureus ATTC 25923 and the soap quality. The results showed that cinnamon, sandalwood, and massoia essential oils contained, respectively, 14, 20, and 9 chemical components, all of which had been identified. The chemical components with the highest percentage contained in the essential oils of cinnamon, sandalwood, and massoia were cinnamaldehyde (91.71%), β-bisabolen (37.79%), and massoia lactone (77.56%), respectively. The soaps’ antibacterial activity against S. aureus ATTC 25923 increased from moderate (with an inhibitory power of 9.67 mm) to strong. The addition of 3% massoia essential oil got the highest inhibitory power of 16.47 mm (strong), followed by that of 3% cinnamon essential oil and 3% sandalwood essential oil, getting 15.37 mm and 12.19 mm, respectively, both of which were considered strong as well. The results of the organoleptic description test showed that each soap had a distinctive color and aroma depending on the type of essential oil added. The quality test results, including transparency, pH, foam stability, water content, and saponification number, indicated that the formulation had good quality.

Effectiveness of Biolarvicides of Imperata cylindrica, Saccharum spotaneum and Andropogon aciculatus on Aedes aegypti larval Mortality and Egg-laying Ability in Adults

Fri, 24 Jan 2025 14:42:44 +0700

Vector-borne disease such as Dengue Hemorrghagic (DHF), transmitted by Aedes aegypti and Aedes albopictus, remain a significant public health concern in Indonesia. Controlling these disease often involves insectides; however, the negative impact of chemical insecticides have prompted interest in organic alternatives derived from plants. Certain weeds, including cogon grass (Imperata cylindrica), wild sugarcane (Saccharum spontaneoum), and needle grass (Andropogon aciculatus), have shown potential as botanical insecticides. Research findings showed that weed root extracts significantly affect larval mortality rate of Ae. Aegypti. At 1000 ppm, larval mortality was significantly higher compared to 100 ppm and the control, while treatments of 1 ppm and 10 ppm showed similar results to the control. Probit analysis revealed that I. cylindrica root extract achieved an LC50 of 974.99 ppm within 24 hours, indicating it could kill 50% of Ae. Aegypti larvae. Within 48 hours, the LC50 dropped to 889.20 ppm. Toxicity tests further revealed significant differences in Ae. Aegypti egg-laying abilities when treated with extracts. Analysis of variance yielded p-values of 0.000 for egg hatching within 72 and 96 hours, highlighting significant differences across samples. These findings suggest the extracts influence mosquito reproduction, warranting further studies to assess the quality of egg hatched from larvae exposed to these treatments. The potential of botanical insecticides derived from weeds represents a promising step toward sustainable mosquito control in the fight against vector-borne diseases.

Antibacterial Activities of Red Mangrove (Rhizophora stylosa Griff.) Leaf Extract against Klebsiella pneumoniae ATCC 700603

Fri, 24 Jan 2025 16:13:45 +0700

Multidrug-resistant (MDR) Klebsiella pneumoniaeis a critical pathogen causing severe human diseases, including pneumonia. Combating the growing threat of MDRK. pneumoniae requires innovative approaches, such as exploring plant-derived antibacterial agents. Rhizophora stylosa Griff., a mangrove species with traditional medicinal uses, is recognized for its bioactive compounds with potential antibacterial properties. However, research on its bioactive constituents remains limited. This study investigated the antibacterial activity of R. stylosa leaf extracts prepared via maceration and liquid-liquid fractionation against K. pneumoniae ATCC 700603. Using agar-well diffusion and cell leakage assays, the water fraction demonstrated moderate inhibition of K. pneumoniae, producing an average inhibition zone of 8.24 mm and a minimum inhibitory concentration (MIC) of 400 mg/ml. UV-Vis spectrophotometry revealed that the water fraction disrupted protein and nucleic acid synthesis, evidenced by leakage of cellular materials at 260 and 280 nm. Additionally, scanning electron microscopy (SEM) images of K. pneumoniae cells treated with the water fraction showed pore formation and structural damage. These results emphasize the antibacterial potential of the water fraction of R. stylosa leaves against MDRK. pneumoniae. Further investigations are necessary to isolate and identify the specific bioactive compounds responsible for these effects. Moreover, comprehensive assessments of activity and toxicity are crucial to advance R. stylosa-based antibacterial agents as promising alternatives for addressing the challenge of MDR bacterial infections.

Molecular Evidence Points to Strong Resemblance in the Parasitoid Species of Rice and Cogongrass Gall Midges, Platygaster spp. (Hymenoptera: Platygasteridae)

Purnama Hidayat, Yuliana Ayu Lestari, Sari Nurulita — Mon, 03 Feb 2025 11:37:05 +0700

The rice gall midge, Orseolia oryzae, and the cogongrass gall midge, O. javanica, cause gall formation on rice and cogongrass (alang-alang) (Imperata cylindrica). Two different species parasitize these two gall midges but closely related platygasterids, Platygaster oryzae on the rice gall midge and P. orseoliae on the cogongrass gall midge. Both the gall midges and their parasitoids are often found in the adjacent area, raising a question about the relationship between the two gall midges and their parasitoids. This research aims to study the molecular identity of the rice and cogongrass gall midges, along with their platygasterid parasitoids, based on partial sequences of the mtCOI gene. Samples were collected from rice and cogongrass in the adjacent area in Cianjur, West Java Province, and a rice field with no cogongrass in Bogor, West Java Province. Successful DNA amplification was achieved using universal primers for mtCOI. Nucleotide sequencing analysis revealed that the rice gall from Bogor and Cianjur shared 100% similarity and 93.2-99.3% similarity with the rice gall from other countries. Notably, the parasitoids P. oryzae collected from rice in Bogor and Cianjur shared 97.2% similarity with P. orseoliae collected from cogongrass in Cianjur. These findings suggest that the platygaster parasitoids associated with the rice gall and the cogongrass gall midges are identical, serving as potential natural enemies for both pests. This study represents the first molecular identification report of rice and cogongrass gall midges and their platygasterid parasitoids from Java Island, Indonesia.

Potential of Cinnamomum burmanni Leaf Extract as an Exogenous Antioxidant and Spermatoprotective for Rattus norvegicus L. Exposed to Polystyrene Nanoplastics

Tue, 04 Feb 2025 09:31:14 +0700

Polytyrene nanoplastics (NPs) (<1 µm) have high toxicity when entered and accumulated in cells. NPs accumulation causes oxidation stress, thus increasing reactive oxygen species (ROS) production, resulting in necrosis or apoptosis, as well as affecting endogenous antioxidant activity, such as superoxide dismutase (SOD) and catalase (CAT). Cinnamomum burmanni plant contains flavonoids, cinnamaldehyde, phen, and olic acid, potentially exogenous antioxidants. The study aims to analyze the potential of C. burmanni leaf extract for SOD and CAT levels, sperm quality, epithellia tubulus thickness, tubulus seminiferus diameter, and number of spermatogenic Rattus norvegicus cells exposed to NPs. Twenty-five male R. novergicus are divided into five groups (n = 5): two controls (without and with NPs) and three groups (combination of NPs and variations in extract concentrations of 100, 200, and 400 mg/kg. The SOD and CAT levels were measured with an ELISA kit. The histology was observed by counting the spermatogenic cells, measuring the epithellia thickness, and tubulus seminiferus diameter. The sperm motility, viability, and sperm count were observed to determine the sperm quality. Leaf extract of C. burmanni treatment with different concentrations not significantly increased SOD levels but significantly decreased the levels of CAT (P<0.05). The addition of C. burmanni leaf extract significantly increased the spermatogenic cell count, epithelia tubulous thickness, sperm viability, and sperm count (P<0.05), while sperm motility and tubulous seminiferous diameter not significantly increased. According to the results, C. burmanni leaf extract has antioxidant potential against the toxic effects of NPS.

Soil Symbiosis Reimagined: Rhizobium and Mycorrhiza Influence on Soybean Performance in Early Oil Palm Ecosystems

Tue, 04 Feb 2025 00:00:00 +0700

The high demand for soybeans in Indonesia is not in line with national soybean production due to the harvest area decreasing every year. To increase national soybean production, the area of IOP (Immature Oil Plantation) one year has potential for soybean development. This study aimed to investigate the effect of Rhizobium spp. and arbuscular mycorrhizal fungi (AMF) inoculation on the physiological capacity, growth, and yield of the soybean plant planted between rows of IOP one-year-old. This research was carried out at PT Perkebunan Nusantara IV (PTPN IV), Adolina Afdeling 1, Serdang Bedagai Regency, North Sumatra, from March 2023 to June 2023. A two-factor Randomized Complete Block Design (RCBD) with four replications for all treatments was conducted in a field experiment. Experimental treatments include: 1) non-inoculated Rhizobium spp. + non-inoculated AMF (R0M0) (Control), 2) non-inoculated Rhizobium spp. + inoculation AMF (R0M1), 3) inoculation Rhizobium spp. + non-inoculated AMF (R1M0) and 4) inoculation Rhizobium spp. + inoculation AMF (R1M1). The results showed that the single arbuscular mycorrhizal fungi (AMF) inoculation treatment significantly enhanced soybean physiology, growth, and yield including relative water content (85.87%), nitrate reductase activity (8.98 µmol NO₂-g BS^-1 hour^-1), stomatal density (775.16 mm), proline (17.76 µmol proline g BS^-1), and ascorbate peroxidase (0.0068 U/min/mg).

Exploring the α-Amylase Inhibitory Potential of Peronema canescens Jack: An In Vitro and In Silico Study

Tue, 04 Feb 2025 00:00:00 +0700

Hyperglycemia in individuals with type 2 diabetes mellitus is primarily driven by the rapid hydrolysis of starch by the enzyme α-amylase in the pancreas and the breakdown of oligosaccharides by α-glucosidase in the intestine. Peronema canescens Jack. (PC) has shown promise as a potential antidiabetic agent. This study aimed to evaluate the total flavonoid, phenolic, and α-amylase inhibitory activity of extracts and fractions derived from PC leaves using both in vitro and in silico approaches. The ethanol extract of PC leaves was fractionated through liquid-liquid extraction using n-hexane, ethyl acetate, and water as solvents. Preliminary phytochemical screening of the extracts and fractions identified the presence of alkaloids, flavonoids, saponins, tannins, and steroids/triterpenoids. The n-hexane fraction exhibited the highest total flavonoid content, averaging 203.37±4.38 mg QE/gram, while the ethyl acetate fraction demonstrated the highest total phenolic content, averaging 147.04±0.79 mg GAE/gram. Furthermore, the ethyl acetate fraction showed the strongest α-amylase inhibitory activity, with an average inhibition rate of 70.38±1.26%. In silico analysis, combined with GC-MS identification, suggested that three compounds, bis(2-ethylhexyl) phthalate, myristyl oleate, and 14 beta H-pregna may contribute to the observed α-amylase inhibitory activity. These findings highlight the potential of PC as a source of natural antidiabetic agents.

Indonesian Herb Extracts Inhibit the Replication of Bovine Respiratory Syncytial Virus: In Vitro Study

Thu, 06 Feb 2025 09:42:39 +0700

Bovine respiratory syncytial virus (BRSV) is highly prevalent in cattle. It is a major viral cause of bovine respiratory disease complex, which is associated with morbidity, mortality and substantial economic impact. Currently available treatments are only symptomatic, but no specific treatments are available for BRSV infection. This study aimed to identify new antiviral agents against BRSV, which could be used to control bovine respiratory disease complex in cattle with Indonesian herb extracts. Ethanol extracts prepared from Indonesian herbs including Andrographis paniculata, Phyllanthus niruri, Curcuma aeruginosa, and Curcuma xanthorrhiza were evaluated for anti-BRSV activity in A549 cells. The cytotoxicity of the herb extracts was evaluated using a CCK-8 cell viability assay. Antiviral activities of the herb extracts were examined using cell activity and cytopathic assays. The effect on virus production was evaluated by qRT-PCR and plaque-formation assays. Extracts of Curcuma xanthorrhiza (125 μg/ml), Andrographis paniculata (250 μg/ml), and Phyllanthus niruri (62.5 μg/ml) inhibited BRSV activity in A549 in pre-, simultaneously-, and post-infection treatment assays, respectively, as measured by the selective index. Reduction of BRSV activities by the herb extracts correlated with inhibition of viral gene expression and inhibition of plaque formation in a concentration- and time-dependent manner. Our findings suggest that these herb extracts have sufficient potency to be used not only as a therapeutic agent but also as a preventive agent to limit BRSV infection.

Development of Multiplex PCR for Simultaneous Detection of Trypanosoma evansi and Equine Piroplasma Infection in Horses

Arifin Nugraha, Umi Cahyaningsih — Tue, 11 Feb 2025 13:46:55 +0700

The identification of equine piroplasmosis and surra disease in infected horses currently presents a significant challenge. Clinical symptoms and blood smears are commonly used to diagnose these two diseases. The objective of this study was to develop a multiplex amplification assay capable of simultaneously identifying all three blood protozoa (equine piroplasma and Trypanosoma evansi) in a single test. The primer pairs used for detecting T. equi, B. caballi, and Trypanosoma evansi were ema-2-t (587 bp), Bc-134 (429 bp), and Rotat 1.2 VSG (151 bp), respectively. The multiplex PCR assay was subsequently evaluated for its detection limit, sensitivity percentage, and specificity using single PCR as the reference standard. The multiplex PCR method demonstrated a sensitivity of 100% for detecting both Theileria equi and Trypanosoma evansi, with all positive samples confirmed by a single PCR. It also achieved 100% specificity for both the parasites. However, for Babesia caballi, while the sensitivity remained at 100%, the specificity was reduced to 66%, indicating some limitations in accurately identifying negative samples. For the multiplex PCR assay, the minimum detectable concentrations were 0.01 ng/µL for both T. equi and B. caballi, whereas Trypanosoma evansi exhibited a detection threshold of 1 ng/µL. When multiplex PCR assays were used to screen blood samples obtained from horses in selected districts of Bogor, only a single sample showed a positive result for T. equi. The sensitivities and specificities achieved by multiplex PCR and single PCR were comparable. Therefore, the multiplex PCR method developed in this study can be effectively applied to diagnose piroplasmosis and surra in horses.

The pipB Gene as Target for Development of Detection Method of Pathogenic Bacteria Salmonella typhi Using Real-time Polymerase Chain Reaction

Tue, 11 Feb 2025 15:29:05 +0700

Salmonella typhi is a bacteria that leads to typhoid fever and one of the causes of death due to bacteria infections. In Indonesia, typhoid fever occurs around 1,100 cases per 100,000 population per year, with a mortality rate of 3.1-10.4%. It's necessary to develop a rapid and accurate detection of Salmonella typhi. The pipB gene of Salmonella typhi has the function of being an autophagia inhibitor in humans. This study aims to develop a detection kit for Salmonella typhi pathogenic bacteria targeting the pipB gene using a pipB primer in confirmation, specificity, and sensitivity tests. The results showed that pipB primer can amplify Salmonella typhi DNA fragment of 196 bp at the optimum annealing temperatures between 54-62°C. Confirmation test with real-time PCR found that the pipB primer pair (pipB-F and pipB-R) amplified at cycle 12.93 and 13.10 (Duplo) with a Tm value of 84.05°C and 84.20°C (Duplo). Based on the difference and average value produced in the confirmation and specificity test, pipB primer has amplified Salmonella typhi DNA at Ct 12.47±0.6 with a Tm value of 83.62°C±0.6. The pipB primer pair (pipB-F and pipB-R) could distinguish target from non-target bacteria based on their cycle threshold (Ct) and melting temperature (Tm) values. The primer design of pipB primer pair (pipB-F and pipB-R) successfully detected Salmonella typhi bacteria with the smallest concentration of 55.78 × 10² CFU equivalent to 3.2 pg/µL. Based on the results, Salmonella typhi pipB primer successfully detected Salmonella typhi bacteria DNA rapidly, specifically, and sensitively using the real-time polymerase chain reaction method.

Genome-wide Screening of Zinc-Responsive Genes in Arabidopsis thaliana for Enhanced Phytoremediation Potential

Wed, 12 Feb 2025 11:15:58 +0700

Zinc contamination presents considerable threats to human health, agricultural practices, and ecological systems. Arabidopsis thaliana has demonstrated potential for use in phytoremediation, which is an environmentally friendly strategy aimed at addressing this type of pollution. This study identified genes associated with phytoremediation in A. thaliana. This identification was achieved through comparative bioinformatics analysis involving 27 plant species. A critical literature review was conducted. This review aimed to collect genes involved in metal uptake and transport across various hyperaccumulator plant species. A database in BLAST format was generated to enable the study of main homologous protein families, including ATP-binding Cassette transporters, acyl-CoA-binding proteins, metallothioneins, and phytochelatin synthases. In silico screening for zinc-binding was performed by means of a web application based on the deep-learning framework called GNINA. The results highlighted the fact that within the phytochelatin synthases-especially PCS1 and PCS2, the protein PCS2 has the highest affinity to zinc. Heavy metal ATPases involved in the vital process of metal transport and homeostasis include HMA1. Zinc transporters responsible for zinc uptake and sequestration include cation exchanger 1CAX1 and IRT2. The information thus provided explains in detail the complex molecular machinery regulating metal detoxification in A. thaliana and provides insights for developing transgenic plants with enhanced phytoremediation capabilities. Future research should target these genes to create plants with improved metal tolerance and accumulation, advancing phytoremediation technologies.

Comprehensive Characterization of Phospholipase C and D Families in Cocoa (Theobroma cacao L.): Identification, Phylogenetics, Gene Structure, and Transcriptomic Insights

Wed, 12 Feb 2025 14:22:11 +0700

This study provides a detailed investigation of the phospholipase C (PLC) and phospholipase D (PLD) gene families in cocoa (Theobroma cacao), focusing on their identification, characterization, and expression patterns. A total of 10 PLC and 12 PLD genes was identified and systematically annotated based on their sequence homology, conserved domains, and functional classification, adhering to established nomenclature. Analysis of physicochemical properties revealed diversity in molecular weights, isoelectric points, and stability parameters, reflecting their structural and functional variability. Phylogenetic analysis classified the genes into distinct subfamilies and highlighted their evolutionary relationships with homologs in Arabidopsis thaliana and rice (Oryza sativa). Gene structure analysis demonstrated significant variation in exon-intron organization, indicating functional specialization and regulatory complexity within these gene families. Expression profiling during cocoa embryo development showed that certain genes, such as TcNPC2, TcPI-PLC5, and TcPLDα1, were highly expressed, while others exhibited stage-specific activity. In response to Phytophthora megakarya infection, several PLC and PLD genes displayed significant changes in expression across different time points and genotypes, including the upregulation of TcPI-PLC2, TcPLDα5, and TcPLDζ2, suggesting their roles in cocoa's stress responses and defense mechanisms. These findings offer new insights into the biological roles of PLC and PLD gene families in cocoa, particularly in growth, development, and stress adaptation, providing a solid foundation for further functional research and potential applications in cocoa improvement programs.

Analysis of Bird Vocal Bandwidth in Relation to Vegetation Structure Variability in Coffee Agroforestry Systems, Jember, Indonesia

Thu, 13 Feb 2025 14:15:10 +0700

Bird vocalizations are crucial for communication and adaptation to environmental changes. As forests are converted to agriculture, agroforestry plays a key role in conserving biodiversity. This study examines vocal bandwidth variations in four bird species-Common Tailorbird (Orthotomus sutorius), Sooty-headed Bulbul (Pycnonotus aurigaster), Olive-backed Sunbird (Cinnyris jugularis), and Small Minivet (Pericrocotus cinnamomeus)-across three coffee agroforestry sites in Jember, Indonesia: Silo, Kluncing, and Kaliwining. The study explores how vegetation structure influences vocal adaptations using sound recordings, vegetation surveys, the Kruskal-Wallis test, and Pearson correlation analysis. Results show significant vocal bandwidth differences in Common Tailorbird and Sooty-headed Bulbul. Common Tailorbird exhibited the highest bandwidth variation in Silo (2.85 kHz), followed by Kluncing (2.30 kHz) and Kaliwining (1.10 kHz) (p<0.001). Similarly, Sooty-headed Bulbul showed significant variation, with the highest bandwidth in Silo (3.1 kHz), Kaliwining (3.5 kHz), and Kluncing (2.7 kHz) (p<0.001). However, bandwidth variation in Olive-backed Sunbird (p = 0.12) and Small Minivet (p = 0.0014) was not significant. Pearson correlation analysis indicated a strong but non-significant relationship between vegetation structure (Importance Value Index) and Sooty-headed Bulbul presence (R² = 0.8999, p = 0.059). These findings suggest that while vegetation influences some species, other factors like resource availability or human activity may also shape vocal adaptations. This study enhances understanding of bird communication in agroforestry, supporting biodiversity conservation and sustainable land management.

Characterization of SNEDDS Formulated with Single Bulb Garlic and its Anti-Inflammatory Effect on 3T3-L1

Thu, 13 Feb 2025 00:00:00 +0700

Inflammation is an early symptom of a disease that reduces the level of health. Single-bulb garlic (Allium sativum L.) is used medicinally as a plant with a broad pharmacological effect, especially anti-inflammatory activity. Self-nanoemulsifying drug delivery systems (SNEDDS) have offered opportunities to improve drug delivery. The current study aimed to characterize SNEDDS-single bulb garlic extract (SBGE) and determine its potential as an anti-inflammatory agent in 3T3-L1 cells. SNEDDS was formulated from tween-80, glycerol, canola oil, and SBGE. The formula characterization is done using droplet size, polydispersity index, zeta potential, physical stability test, and antioxidant assay. The cytotoxicity test of SNEDDS-SBGE was evaluated using a 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. The anti-inflammatory activity was examined using 3T3-L1 cell methylglyoxal (MG) induction, and the expression of cytokines was measured using immunocytochemistry (ICC). The SNEDDS-SBGE had a nanoemulsion size of 42.30±1.39 nm, 0.6±0.03 for the polydispersion index, and -22.63±0.75 mV for the zeta potential. SNEDDS-SBGE was physically stable and had a high antioxidant level (47.579±8.017 %). SNEDDS-SBGE exhibited no toxic effect on 3T3-L1 cells. The administration of 62.5 µg/ml and 125 µg/ml SNEDDS-SBGE could inhibit (p<0.05) the expression of IL-1β after methylglyoxal induction. Thus, SNEDDS-SBGE may have potential anti-inflammatory properties.

Exploration of Lipid Profile and Wax Ester Content from Local Strain Euglena sp. IDN33 Cultivated in Mixotrophic Condition Combined with Molasses Supplementation

Fri, 14 Feb 2025 11:28:23 +0700

The transition from fossil fuels to sustainable renewable energy is currently growing rapidly. Euglena sp. can be the source of biofuel. Molasses, in addition to Euglena sp., can increase the levels of paramylon and wax esters produced. Therefore, this study aims to analyze cell growth, lipid profile, and wax ester content of Euglena sp. cultivated under mixotroph conditions with molasses. Euglena was cultivated under mixotrophic conditions with the addition of various concentrations of molasses (0, 10, 15, and 20 g/L) until the final log phase. Then, measurements were made on cell density, levels of lipids, proteins, and paramylon to measure the growth and metabolites profile of Euglena sp., along with lipid profile and wax ester analysis using GC-MS. The results of the study showed that 10 g/L molasses increased Euglena growth (SGR 0.1736± 0.0213 mg/ml) and carbohydrate (0.426 mg/ml) but reduced lipid accumulation (2.29 mg/ml). This treatment provides the best result before control. Molasses does not increase carbohydrates and protein but increases paramylon accumulation. The GC-MS assay detected more lipid profiles and concentrations of each lipid type in non-molasses-treated Euglena sp. compared to molasses-treated. Euglena was cultivated on molasses, which was dominated by palmitic acid and myristyl myristate with the absence of PUFA-type fatty acids in the cells, while the untreated ones were dominated by stearic acid and myristyl myristate. Thus, 10 g/L supplementation of molasses is the most effective treatment compared to others.

Oxidation Stress of UV C Light on Growth, Carotenoids and Chlorophyll-a Content of Chlorella vulgaris

Fri, 14 Feb 2025 13:40:57 +0700

C. vulgaris is photoautotrophic, which refers to microalgae that require light as an energy source. One of the light sources used to affect pigments in microalgae is UV light. The advantage of UV light is that it can change gene composition, which results in mutant genes in biopigment mutations and causes a higher number of activities compared to the natural ones, thereby increasing the growth, carotenoids, and chlorophyll-a of microalgae. The aim of this research is to determine the effect of oxidation stress of UV light on growth, carotenoids and chlorophyll-a content of Chlorella vulgaris. The treatments were given based on differences in the power of the UV-C light from, 8W, 15W and 30W. This study used the UV-Vis Spectrophotometer method was used to calculate growth, carotenoids and chlorophyll-a. Data were analysed using ANOVA test, Duncan's Multiple Range Test, and Principal Component Analysis (PCA). The results showed a significant effect of UV-C light. The best treatment was found in the administration of 30 W UV light, with the results of growth was 4.7746 × 10⁶ cells/ml, content of carotenoids was 1.75 mg/ml and chlorophyll-a was 4.67 mg/ml of C. vulgaris. Biopigment can absorb radiation from UV-C light, which causes these microalgae to survive in environmental stress conditions. Absorbed light can affect the spectrum of pigments, thereby increasing growth, carotenoids, and chlorophyll-a.

The Ureolytic Soil Bacteria Bacillus albus, a potential Agent for Biocement

Fri, 14 Feb 2025 14:07:52 +0700

Concrete is a common building material and is very vulnerable to cracking caused by unstable temperature/humidity. Concrete crack repair can be done by using microorganism substitution that can produce CaCO₃ (calcite) compounds that can be used as an environmentally friendly method in improving structural formation and increasing the strength and durability of concrete, one of which is using ureolytic bacteria. This study aimed to isolate and characterize ureolytic bacteria isolates and then to assess the calcite precipitation potential of ureolytic bacteria isolates from landfills. The ureolytic bacterial isolates were grown on NB-U/Ca and tap water medium. Analysis of Calcite Structure using Fourier Transform Infra-Red (FTIR), and molecular identification using 16S rRNA gene sequences. Bacterial isolate SP. 48 were able to grow and produce calcite in both media, especially in tap water medium. FTIR results showed that the precipitates produced by bacterial isolates on both mediums had strong absorption peaks, which were detected to be calcite. Molecular identification using the 16S rRNA gene sequence showed that the isolate is Bacillus albus. B. albus is a proteolytic bacterium collected from landfills that was proven to be a calcite-producing bacterium, a new finding in this study. B. albus can grow and produce calcite in a tap water medium with low pH. This finding can be used as an alternative to overcome concrete cracks and increase the strength and durability of concrete. This bacterial isolate could be developed as a biocement candidate.