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Molecular Detection of Eimeria bovis in Indonesian Beef Cattle Using Nested PCR Technique Nasrulloh, Mukh Fajar; Nurcahyo, Raden Wisnu; Priyowidodo, Dwi; Ekawasti, Fitrine; Firdausy, Lintang Winantya
HAYATI Journal of Biosciences Vol. 31 No. 5 (2024): September 2024
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.31.5.980-987

Abstract

Eimeria bovis is a pathogenic protozoan that causes cattle digestive tract infections, which can cause economic losses to farmers. It is necessary to develop specific and accurate detection methods to conserve livestock and prevent coccidiosis in Indonesia. This study aims to detect E. bovis by nested PCR and determine the relationship with reference sequences. A total of 167 samples of beef cattle feces were taken randomly from community farms spread across 18 provinces in Indonesia. The feces were examined natively, and then the oocysts were purified by the sugar flotation method, extracted by KIT extraction, and amplified by the nPCR technique. Positive samples were followed by sequencing and phylogenetic analysis using MEGA 11 software. This study used two pairs of primers (outer and inner) taken from ITS-1 molecular markers. As many as 96 out of 167 samples (57.5%) were positive for Eimeria spp., and 48 of the 96 samples were positive for Eimeria spp. (50%) were detected to be positive for E. bovis based on the presence of a 238 bp DNA fragment. The results of the phylogenetic analysis showed that the study sample formed a separate cluster from the E. bovis cluster from abroad. In conclusion, E. bovis was detected in 16 out of 18 provinces in this study, and the nPCR technique proved to have better sensitivity and specificity.
Studi Literatur: Faktor Risiko dan Strategi Pengendalian Fasciolosis di Indonesia Charisma, Acivrida Mega; Ekawasti, Fitrine; Wahyuningrum, Arina; Naisali, Walburga F. F.; Syawaludin, Moch. Fardha; Auralya, Theresia Feby; Vebiayanti, Saski Dwi; Maisaroh, Maisaroh; Rukmana, Dinda Herdiana; Alfani, Moh.; Afkarina, Zida Kamalia
JURNAL KAJIAN VETERINER Vol 12 No 2 (2024): Jurnal Kajian Veteriner
Publisher : FAKULTAS KEDOKTERAN HEWAN UNIVERSITAS NUSA CENDANA

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.35508/jkv.v12i2.19337

Abstract

Fasciola hepatica is the main species found infecting livestock. Fasciolosis is a significant animal health and public health problem, causing major economic losses. Epidemiological studies of fasciolosis in dairy cattle in Indonesia are still few and existing reports only focus on prevalence. Fasciolosis is an endemic parasitic disease in Indonesia and causes considerable economic losses. Efforts that can be made to prevent disease are vector eradication, namely eradicating snails through biological means, for example by raising ducks. Raising ducks biologically can reduce the worm population. Several simple techniques for controlling worm infections in cattle can be done by regulating feeding, timing grass cutting and making compost to break the life cycle of Fasciola sp. In general, dairy cattle in Indonesia are raised more intensively compared to beef cattle farms, which explains the difference in prevalence rates. In intensive farming systems, animals are housed permanently, limiting their exposure to parasite-contaminated grass compared to pastured animals.
Anthelmintic Potential of Annona muricata seed Infusion: An In Vitro Study on Haemonchus contortus in Goats Baihaqi, Zein Ahmad; Lisnanti, Ertika Fitri; Tamara, Safira; Ekawasti, Fitrine; Anggrahini, Santika; Fauziah, Ima
Jurnal Sain Veteriner Vol 43, No 1 (2025): April
Publisher : Faculty of Veterinary Medicine, Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.100652

Abstract

Parasitic infections, particularly those caused by Haemonchus contortus, significantly impact the health and productivity of small ruminants worldwide. Traditional anthelmintic treatments are increasingly challenged by resistance issues, prompting the search for alternative treatments. The objective of this study was to assess the effectiveness of Annona muricata seed infusion on Haemonchus contortus mortality in goats in vitro. The research was conducted from November 2023 to March 2024 at the Laboratory of the Faculty of Agriculture, Islamic University of Kadiri, and BRIN. The method employed was one-way ANOVA. Treatments included a negative control with 0.9% NaCl, 10% concentration infusion, 20% concentration infusion, and albendazole. Mortality rates were assessed at 0.5, 1, 2, 4, 6, 8, 10, and 12-hour intervals. The results revealed that Annona muricata seed infusion contains compounds such as tannins, flavonoids, alkaloids, saponins, and steroids, which are capable of damaging the cuticle and buccal tissues of Haemonchus contortus. Phenolic content was measured at total phenolics of 9.1 mg RE / g Dw, total flavonoids of 4.7 mg RE / g Dw, total tannins of 7.2%, condensed tannins / CT of 3.4%, and hydrolyzable tannins / HT of 3.9%. Infusions at 10% and 20% concentrations achieved 100% mortality of Haemonchus contortus by the 8th hour. Scanning Electron Microscopy (SEM) revealed structural changes in Haemonchus contortus at different infusion concentrations, including wrinkling, irregular lines, and the presence of holes in the cuticle and buccal regions. The infusion of Annona muricata seed infusion is effective as an anthelmintic against Haemonchus contortus, achieving 100% mortality at concentrations of 10% and 20% within 8 hours. The study also demonstrates structural damage to the worms, affirming the potential of Pineapple Peel infusion as a natural parasitic control in small ruminants.
Studi Literatur: Efektivitas Deteksi Malaria Menggunakan Pemeriksaan Mikroskopis Apusan Darah Charisma, Acivrida Mega; Ekawasti, Fitrine; Azis, Mutiara Zakhilah; Sari, Adela Permata; Suci, Azlin Eva; Putra R, Rafi; Putri, Sintia Cahyani; Sugiarto, Agus Bambang
Jurnal Kedokteran Universitas Palangka Raya Vol 13 No 1 (2025): Jurnal Kedokteran Universitas Palangka Raya
Publisher : Fakultas Kedokteran, Universitas Palangka Raya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37304/jkupr.v13i1.17393

Abstract

Malaria is an infectious disease caused by the parasites Plasmodium malariae, Plasmodium vivax, Plasmodium, falciparum, Plasmodium ovale, and Plasmodium knowlesi. The purpose of the literature review is to determine the effectiveness of malaria detection microscopically using blood smears. This research method uses data from scientific publications sourced from PubMed, ScienceDirect, and Google Scholar. Malaria cases in 2019-2021 have increased worldwide. Deaths from malaria that occurred worldwide in 2019-2020 increased by 12%. Indonesia contributes the second largest number of malaria cases in the Asian region after India, especially in the eastern region including Papua Province, West Papua Province, East Nusa Tenggara Province, and Maluku Province. Mortality rates from malaria still occur in several provinces in Indonesia. The purpose of this study is to determine the effectiveness of malaria detection using microscopic examination of blood smears. This research method is a literature review of the description of research results that have been conducted by others. Searches on the Google Scholar and Science Direct search engines use boolean techniques. This journal review was taken because the detection of malaria using blood smear microscopic tests is one of the most commonly used common methods, and can help improve professional competence in diagnosing and planning appropriate treatment for malaria sufferers, so it is relevant for doctors, analysts, and health researchers. Blood smear microscopy is the main way to detect malaria using thick blood preparations, this test can find Plasmodium malariae, Plasmodium vivax, Plasmodium, falciparum, Plasmodium ovale, Plasmodium knowlesi even in small amounts. Thin blood preparations help identify parasite species accurately.  
Hubungan Perilaku Hidup Bersih Sehat Dengan Kejadian Infeksi Kecacingan Soil Transmitted Helminth Di Daerah Tempat Pemrosesan Sampah Terpadu: The Relationship Of Clean Healthy Living Behavior With The Incidence Of Soil Transmitted Helminth Worm Infections In The Integrated Waste Processing Place Mega Charisma, Acivrida; Aini, Jessica Dewi Nur; Puspita, Chelsea Dhea; Chairunnisa, Adellia; Yulianda, Siska; Klau, Adrigius Neno; Ekawasti, Fitrine
Binawan Student Journal Vol. 7 No. 1 (2025)
Publisher : Direktorat Penelitian, Pengabdian Masyarakat, Dan Kerjasama Universitas Binawan (DPPMK Universitas Binawan)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.54771/r10nw827

Abstract

Infeksi kecacingan Soil Transmitted Helminths (STH) prevalensi yang cukup tinggi di Indonesia terutama di daerah yang memiliki lingkungan sanitasi buruk. Penyebab STH terbanyak di Indonesia adalah Ascaris lumbricoides dan Trichuris trichiura. Perilaku hidup bersih sehat (PHBS) mencuci tangan, mengonsumsi air bersih, serta kebersihan lingkungan, dapat memengaruhi tingkat kejadian infeksi kecacingan. Potensi kecacingan di Desa Jabaran sangat tinggi karena kondisi lingkungan Tempat Pemrosesan Sampah Terpadu (TPST) yang belum sepenuhnya memenuhi standar sanitasi dan kebersihan. Penulisan artikel ini bertujuan untuk mengevaluasi kaitan antara perilaku PHBS dan kejadian infeksi STH di TPST Desa Jabaran. Jenis penelitiannya adalah studi literatur. Artikel ini menunjukkan bahwa kurangnya perilaku PHBS meningkatkan risiko infeksi STH di daerah tersebut, dengan tingkat kejadian yang lebih tinggi di masyarakat yang tidak menerapkan kebiasaan hidup sehat. Faktor lingkungan TPST yang terkontaminasi juga berkontribusi pada penyebaran infeksi kecacingan.
Genomic Characterization of Antimicrobial Resistance Genes in Pasteurella multocida Isolates from East Nusa Tenggara and Lampung, Indonesia Prihandani, Sri Suryatmiati; Wibawan, I Wayan Teguh; Noor, Susan Maphilindawati; Ekawasti, Fitrine; Khairullah, Aswin Rafif; Purba, Hastuti Handayani S.; Puarada, Alif Rahman Rohim; Safika, Safika
Jurnal Medik Veteriner Vol. 8 No. 2 (2025): October
Publisher : Universitas Airlangga

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.20473/jmv.vol8.iss2.2025.368-381

Abstract

Haemorrhagic Septicaemia (HS) is a devastating disease affecting cattle and water buffaloes in Indonesia, causing annual economic losses in livestock industries, particularly in Asia, Africa, and the Middle East. Pasteurella multocida, the causative agent of the disease, has shown increasing antimicrobial resistance, complicating treatment efforts. Therefore, this study aims to provide the first genomic analysis of P. multocida isolates from different Indonesian provinces, focusing on genes conferring resistance to beta-lactam antibiotics based on Whole Genome Sequencing (WGS) results. Genomic data can be used to confirm the results of phenotypic antibiotic resistance testing. P. multocida isolates analyzed in this study were sourced from the Indonesian Research Center for Veterinary Science (IRCVS) collection, which included 2 samples originating from different geographic locations within Indonesia. The samples were subjected to biochemical, molecular, and antimicrobial susceptibility testing. WGS was performed using Oxford Nanopore Technologies (ONT) with subsequent bioinformatics analysis for genome assembly and resistance profiling. Phenotypic analysis showed significant variations between the isolates. The NTT isolate showed resistance to Penicillin, while the Lampung 2952 remained susceptible in the disk antibiotic test. Genome sequencing revealed extensive resistance determinants, including β-lactamase genes (blaZ, blaR1) in the NTT isolate, correlating with its phenotypic resistance. The high-quality genome assemblies (N50: 17,225 bp for NTT; 12,662 bp for Lampung 2952) enabled comprehensive resistome characterization, identifying more than 22 resistance genes in each isolate, including novel variants not previously reported in Indonesian strains. Therefore, this study provides the first genomic and resistance analysis based on Whole Genomic Sequencing data of Indonesian P. multocida isolates. The results show the urgent need for enhanced surveillance and prudent antimicrobial use in livestock management.
Detection of Very Virulent Infectious Bursal Disease (IBD) in Chicken in West Java Putri, Ajeng Fabeana; Winarsongko, Agus; Hoerudin, Heri; sekarmila, Gita; Ahpas, Ahpas; Jaelani, Jejen; Gunawan, Wawan; Dewiyanti, Rina; Pratama, Yuda; Nuradji, Harimurti; Fairusya, Nuha; Ekawasti, Fitrine; Adji, Rahmat Setya; Dharmayanti, NLP Indi; Indriani, Risa; Utomo, Bambang Ngaji; Suryatmiati, Sri
Jurnal Medika Veterinaria Vol 18, No 1 (2024): J.Med.Vet.
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.med.vet..v18i1.35819

Abstract

Infectious Bursal Disease (IBD), also known as Gumboro disease, is an acute, highly contagious disease that infects chickens and causes a high mortality rate of up to 100% in young animals. The disease is caused by Infectious Bursal Disease Virus (IBDV) of the genus Avibirnavirus, family Birnaviridae. The disease has been reported in Indonesia since 1976, and management strategies for the disease, such as vaccination, have been applied to prevent and control outbreaks in poultry farms. In this study, we conducted the detection of the disease in chickens from a farm in West Java with a mortality rate of 80%. Chickens showing clinical signs, such as sudden death, anorexia, watery diarrhea, and ruffled feathers, were necropsied, and organ samples, including the bursa Fabricius, brain, and spleen, were collected. The samples were then tested using Reverse-Transcriptase Polymerase Chain Reaction (RT-PCR) to confirm the diagnosis of IBD. Positive results were obtained in this study, highlighting the need for improved biosecurity in poultry farms in Indonesia. These results also provided a basis for further research on viral characterization to develop detection kits or vaccines for IBD using local isolates from the field in Indonesia.
EVALUATION OF B1 GENE TO DETECT Toxoplasma gondii: COMPARISON OF THREE SETS NESTED PCR PRIMER Ekawasti, Fitrine; Azmi, Zul; Subekti, Didik Tulus; desem, muhammad ibrahim; Nugraha, Arifin Budiman; Sadiah, Siti; Cahyaningsih, Umi
Jurnal Kedokteran Hewan Vol 17, No 2 (2023): June
Publisher : Universitas Syiah Kuala

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21157/j.ked.hewan.v17i2.22251

Abstract

This study aimed to evaluate three sets of B1 gene DNA primer for the diagnosis of Toxoplasma gondii. The DNA of Toxoplasma gondii that stored on liquid nitrogen was isolated using DNAzol reagent. The first step of Polymerase Chain Reaction (PCRs) was performed using external and internal primer sets, respectively, and then nPCR. PCR products sequencing was performed by Apical Science. All sequences were analysed using CLC Sequence Viewer Version 8.0 software and compared to sequence database that deposited in ToxoDB (Toxoplasma gondii genome database) using BLAST (https://toxodb.org/toxo/app). Each B1 gene primer was evaluated by performing single PCR (forward and reverse) and nested PCR reactions. Three sets of B1 gene primer have different amplification precision. According to the results of amplicon sequencing, the primer set #2 has the best amplification precision of B1 gene.
Optimization of Sybr Green Quantitative Real Time Polymerase Chain Reaction (qPCR) using Excreted-Secreted Antigens (ESAs) Genetik Marker for Detection Toxoplasma gondii Ekawasti, Fitrine; Winarsongko, Agus; Nepho, Farlin; Purwanto, Eko Setyo; Subekti, Didik Tulus; nuradji, harimurti; Dharmayanti, NLP Indi; Ahmad, Riza Zainuddin; Sa’diah, Siti; Cahyaningsih, Umi; Nurcahyo, Raden Wisnu
Jurnal Sain Veteriner Vol 42, No 1 (2024): April
Publisher : Faculty of Veterinary Medicine, Universitas Gadjah Mada bekerjasama dengan PB PDHI

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.22146/jsv.90867

Abstract

AbstractToxoplasma gondii is an obligate intracellular parasite, causing toxoplasmosis in almost all warm-blooded animals and humans worldwide. Toxoplasmosis is a zoonotic disease of serious public health concern. Host cell invasion by T. gondii tachyzoites has process involving the sequential secretion of Excreted-Secreted Antigens (ESAs). T. gondi ESAs could be a valuable candidate for the diagnosis of toxoplasmosis. Techniques to more accurately detection of T. gondii recently developed biotechnological methods that are currently being used, conventional and real time Polymerase Chain Reaction (RT-PCR). RT-PCR is more widely used because it is more sensitive and specific. The aims of this study were to optimize the Sybr Green RT-PCR in different region gene based on Excreted-Secreted Antigens (ESAs), tachyzoite surface antigen and bradhyzoite antige, then adapt the conventional PCR program to real-time PCR for detection Toxoplasma gondii. Optimization is necessary to get optimal condition of PCR to get the best results. T. gondii RH strains derived from liquid nitrogen and DNA extracted by DNAzol. The genetic marker used GRA1#1, GRA1#2, GRA7#1, GRA7#2, ROP1, MIC3, SAG1 and BAG1. The results of the optimization of multiple primer genes can adapt and be used optimal in RT-PCR by using the same cycle program simultaneously in one run. Overall, RT-PCR for the detection of T. gondii DNA demonstrated excellent agreement with conventional PCR. RT-PCR with melting curve analysis is rapid and simple that facilitates high throughput analysis to detect T. gondii. The optimal conditions obtained from the optimization results can facilitate further research to detect T. gondii.Keywords: Biotechnology molecular, Detection, excretory-secretory antigen, toxoplasmosis