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Development of Multiplex PCR for Simultaneous Detection of Trypanosoma evansi and Equine Piroplasma Infection in Horses Nugraha, Arifin; Cahyaningsih, Umi
HAYATI Journal of Biosciences Vol. 32 No. 3 (2025): May 2025
Publisher : Bogor Agricultural University, Indonesia

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.4308/hjb.32.3.740-746

Abstract

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.
Pemberian Ekstrak Air Bidara Laut (Strychnos Ligustrina) terhadap Diferensial Leukosit Mencit yang diinfeksi dengan Plasmodium berghei Nurridho Wahid, Muhammad; Cahyaningsih, Umi; Nugraha, Arifin
Jurnal Veteriner dan Biomedis Vol. 2 No. 1 (2024): Maret
Publisher : Sekolah Kedokteran Hewan dan Biomedis

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/jvetbiomed.2.1.35-42.

Abstract

Plasmodium berghei merupakan hemoprotozoa penyebab penyakit malaria pada mencit. Tujuan penelitian ini untuk mengetahui pengaruh ekstrak air bidara laut terhadap gambaran diferensial leukosit pada mencit yang diinfeksi Plasmodium berghei. Penelitian ini menggunakan 49 ekor mencit Balb/c, dibagi menjadi 7 kelompok yaitu kontrol normal (KN), kontrol infeksi tanpa perlakuan (KI), kontrol infeksi yang diberikan perlakuan obat kombinasi dihydroartemisinin 25 mg/kgbb dan piperakuin fosfat dosis 197 mg/kgbb (K0), EAa (ekstrak aquades), EAb (ekstrak aquades : DHF = 1:1), EAc (ekstrak aquades : DHF = 1:2), EAd (ekstrak aquades : DHF = 2:1). Kecuali KN, semua kelompok mencit diinfeksi Plasmodium berghei sebanyak 1 x 10 6 /ml secara intraperitoneal. Setiap hari diambil darah dari ekor mencit, dibuat apusan tipis dan dilakukan pewarnaan dengan Giemsa 10%, kemudian dilakukan perhitungan diferensial leukosit. Nilai rata-rata persentase diferensial leukosit diolah menggunakan uji ANOVA dan perbedaan hasil persentase pada masing-masing kelompok diketahui dengan menggunakan uji Duncan menggunakan software SPSS. Hasil penelitian menunjukkan bahwa perlakuan pada kelompok EAa dan EAd mengalami penurunan persentase neutrofil dan meningkatkan persentase limfosit pada hari ke 6 setelah infeksi. Persentase monosit, eosinofil dan basofil tidak mengalami perubahan yang signifikan. Pengobatan ekstrak air bidara laut dan DHF berpotensi sebagai obat antimalaria dengan menurunkan tingkat parasitemia dan meningkatkan persentase limfosit.
Pendampingan Pengeringan Tanaman Obat dengan Tipe Fluidized Bed Dryer untuk Usaha Mikro, Kecil dan Menengah di Kota Bogor Muhandri, Tjahja; Nugraha, Arifin; Cahyaningsih, Umi
Agrokreatif: Jurnal Ilmiah Pengabdian kepada Masyarakat Vol. 9 No. 2 (2023): Agrokreatif Jurnal Ilmiah Pengabdian Kepada Masyarakat
Publisher : Institut Pertanian Bogor

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.29244/agrokreatif.9.2.242-247

Abstract

Drying with the fluidized bed dryer type is a drying process by blowing hot air from the bottom onto a stretch of material, so that the material scatters to resemble fluid properties. The drying method with dry air is used to speed up the drying process and maintain the quality of the dry ingredients. The purpose of this service is to provide theoretical and practical drying training to the micro, small and medium enterprises (MSME) group so that they are able to produce dry materials of good quality, namely green color and low water content so they are not easily damaged and can last longer. The assistance was carried out for Naphyco MSME partners and representatives of RW 10 residents, Tanah Baru Village, North Bogor District, Bogor City in October 2022 at Naphyco MSME partner locations. The training was carried out using two methods, namely presentation and hands-on practice of drying medicinal plants with a fluidized bed dryer. The positive impact of this assistance is to gain knowledge about how to dry medicinal plants faster than drying them in direct sunlight, and also to obtain leaves of medicinal plants that are green in color (not brown). The achievement of mentoring activities can be seen from the results of the pre-test and post-test. The results of the questionnaire showed an increase in participant knowledge from a good score of 55.6% to 88.9%, a moderate score from 22.2% to 0% and a value less than 22.2% to 11.1%. There was also an increase in participants' interest in the questionnaire statements who strongly agreed from 66.7 to 88.9%, who agreed from 33.3% to 11.1%. The results of the questionnaire above need to be continued with a mentoring program for other MSME members to produce quality products as raw materials.