Evaluasi Validitas Human Cortisol Enzyme-Linked Immunosorbent Assay (ELISA) Kit dan Waktu Sentrifugasi Sampel Darah untuk Pengukuran Konsentrasi Hormon Kortisol pada Kambing Kacang

ABSTRAK. Penggunaan human cortisol enzyme-linked immunosorbent assay (ELISA) kit untuk pengukuran hormon kortisol pada hewan dan keterlambatan waktu sentrifugasi sampel darah untuk analisis hormon perlu dievaluasi. Penelitian ini bertujuan untuk mengevaluasi validitas human cortisol ELISA kit (EIA-1887, DRG Instruments GmbH, Germany) untuk pengukuran konsentrasi kortisol dan menguji pengaruh keterlambatan waktu sentrifugasi sampel darah terhadap stabilitas konsentrasi kortisol pada kambing kacang. Sampel darah dikoleksi dari delapan ekor kambing kacang. Uji validitas kit EIA-1887 dilakukan secara: a) analitik (uji parallelism, akurasi, dan presisi), dan b) biologis (pengukuran kortisol sebelum dan setelah transportasi). Uji keterlambatan waktu sentrifugasi terhadap stabilitas konsentrasi kortisol dilakukan dengan 5 perlakuan yaitu disentrifugasi kurang dari 1 jam (P1/kontrol), 6 jam (P6), 12 jam (P12), 18 jam (P18),dan 24 jam (P24) setelah darah dikoleksi. Data uji parallelism dianalisis dengan uji persamaan kemiringan, uji presisi dihitung % CV (coefficient variation) intra-assay dan inter-assay, uji akurasi dihitung % recovery, uji T untuk validasi biologis, dan uji ragam (One Way Anova) untuk pengaruh waktu sentrifugasi. Hasil uji parallelism menunjukkan kurva sampel kambing kacang sejajar/parallel dengan kurva standar kortisol. Akurasi kit EIA-1887 adalah 103,43±7,85%, dan % CV intra-assay dan inter-assay adalah 10%. Konsentrasi kortisol setelah transportasi secara signifikan lebih tinggi daripada sebelum transportasi (p0,05). Adanya penurunan secara nyata konsentrasi kortisol pada darah yang disentrifugasi 24 jam (P24) setelah koleksi (p0,05). Kesimpulan, human cortisol ELISA kit (EIA-1887) memiliki validitas yang baik secara analitik dan biologis untuk digunakan dalam pengukuran konsentrasi kortisol pada kambing kacang. Keterlambatan waktu sentrifugasi selama 24 jam berpengaruh terhadap konsentrasi kortisol.  (Evaluation the Validity of Human Cortisol Enzyme-Linked Immunosorbent Assay (ELISA) Kit and Centrifugation Time of Blood Sample for Measuring the Concentration of Cortisol in Kacang Goats)  ABSTRACT. The use of human cortisol ELISA kit for measuring cortisol in animals and delayed to blood centrifugation time for hormone measurement need to be evaluated. This study aimed to evaluate the validity of human cortisol ELISA kit (EIA-1887, DRG Instruments GmbH, Germany) for cortisol measurement and effect of delayed to blood centrifugation time on cortisol concentrations in kacang goats. Blood was collected from eight kacang goats. Validation test of EIA-1887 kit was performed through: a) analytical (parallelism, accuracy, and precision tests), and b) biological validations (measuring cortisol concentrations before and after transportation). Five treatments were performed to test delayed to centrifugation time: blood centrifuged at 1 h (control, P1), 6 h (P6), 12 h (P12), 18 h (P18), and 24 h (P24) after collection. Parallelism data were analyzed by slope equality test, precision and accuracy calculated by % CV of intra-and inter-assay, and % recovery, respectively. Data of biological validation and centrifugation time effects were analyzed by Student t-test, and one way ANOVA, respectively. Results of parallelism showed that serial dilution curve of kacang goat plasma was parallel with cortisol standard curves. Accuracy of EIA-1887 kit was 103.43±7.85%, and % CV of intra-and inter-assay were 10%. Concentration of cortisol after transportation was significantly higher than before transportation (p0.05). Concentration of cortisol was significantly decreased when blood was centrifuged at 24 h after collection (P0.05). In conclusion, human cortisol ELISA kit (EIA-1887) is a reliable assay for measuring cortisol in plasma of kacang goat. Delayed to blood centrifugation time affect cortisol concentrations

CHARACTERISTIC OF OVARIAN AND ESTRADIOL CONCENTRATIONS IN THE FOLLICULAR FLUID OF SLAUGHTERED ACEH CATTLE

The objective of this study was to characterize the morphology of ovary from slaughtered Aceh cow and measuring the concentration of estradiol in the follicular fluid to examine the potential utilizing for in-vitro fertilization (IVF) development in Aceh cattle. In total, 30 pairs of ovaries were collected from 30 Aceh cattle slaughtered at Banda Aceh abattoir. Morphological characteristics of ovaries observed were the size of ovaries, weight of ovaries, number of follicles, the diameter of follicles, corpus luteal (CL), and volume of follicular fluid. The follicular fluid was aspirated using a disposable syringe and analyzed the estradiol concentrations using enzyme-linked immunosorbent assay (ELISA). The follicles were grouped into 3 categories based on the diameter (Ø) of follicles: small follicles (Ø 1 - 5 mm), medium follicles (Ø 5 - 8 mm), and large follicles (Ø 8 mm). The morphological characteristic of ovaries varied between right and left ovaries in which right ovaries showed bigger size and weight compared to the left ovaries (P0.05), whereas the number of follicles and follicular fluid did not significantly different (P0.05). The concentrations of estradiol in follicular fluid of large and medium follicles were significantly higher compared to small follicles (P0.05), whereas the concentration of estradiol between large and medium follicles, did not significant different (P0.05). In conclusion, the follicles from abattoir can be used as potential oocytes resources to support IVF development for Aceh cattle in the future. The follicular fluid provided from these follicles can be an alternative for in vitro maturation media of oocytes due to containing high estradiol concentrations

Concentration of B-CG and sFlt-1 in Rattus Norvegicus Model of Preeclampsia with Swimming Exercise Treatment

Preeclampsia (PE) is a life-threatening pregnancy complication for the mother and fetus. High concentrations of human chorionic gonadotrophin (hCG) and soluble fms-like tyrosine kinase-1 (sFLt-1) during pregnancy may have a role in the pathophysiology of PE. Swimming Exercise (SE) is one of the physical activities recommended for pregnant women and carries a minimal risk. This study is aimed at analyzing the interaction between the conditions of rats (normal and PE), the onset of PE (early onset and late onset), and the time of SE (SE 0 minutes; SE 5 minutes; SE 10 minutes) on the concentrations of B-CG and sFlt-1 in the Rattus norvegicus (R. norvegicus) model of PE. 72 R. norvegicus were included in this study and divided into 12 experimental groups (each group n = 6 individuals). R. norvegicus PE was prepared by inducing L-Nitro-Arginine-Methyl Ester (L-NAME) at a 75 mg/kg BW/day dose. The determination of PE was supported by the observation of differences in the values of urine protein (PU), urine glucose (GU), and urine leukocytes (LU) in R. norvegicus before and after injection of L-NAME. The three-factorial statistical test showed a significant interaction between the concentration of B-CG and the condition of R. norvegicus, the onset of PE, and the time of SE, with a p-value <0.001. The three-factorial statistical test also showed a significant interaction between the sFLt-1 concentration and the condition of R. norvegicus, the onset of PE, and the time of SE with p<0.05. The difference in the concentration of B-CG and sFLt-1 R. norvegicus in each treatment group was influenced by the condition of the rats (normal and PE), the onset of PE (early onset and late onset), and the time of SE (SE 0 minutes; SE 5 minutes; SE 10 minutes). Research related to SE on PE still needs to be continued to decide on recommendations on whether SE can be used as a preventive measure in complementary midwifery care for preventing and reducing symptoms of PE in pregnancy. Doi: 10.28991/ESJ-2023-07-03-021 Full Text: PD

Penataan Permukiman Di Kawasan Kampung Seng Kota Blitar

Arrangement of Slum in a dense residential neighborhood that tends to be slum must fulfill basic principles. First, legalization of ownership status on land and housing sites, Second, provision or improvement of technical services, such as clean water, waste and management of liquid waste, sanitation, electricity, road sidewalks, street lighting. Third, provision or improvement of social infrastructure such as schools, clinics, community centers, playgrounds, green areas. Fourth, the improvement of the physical environment, including the rehabilitation / improvement of Ineligible Houses. Fifth, if deemed necessary, the construction of new housing units is included in the up-grading scheme. Sixth, a redesign of the settlement development plan, including the harmony of buildings and road patterns in accordance with infrastructure needs. Seventh, a change in the policy or regulatory framework that is more in line with the needs and opportunities available to the urban community. However, the involvement of people who live in slum areas, from the planning process to the implementation of the arrangement and improvement of the quality of slum areas, is a must. So that in determining the priority of community activities will feel part of the improvement and development in the region. From the participation and empowerment, there will be a response from the residents' expectations of the living conditions in accordance with their needs and increased comfort. For this reason, the paradigm from development to empowerment of the development and construction of settlements, requires the comfort of living to be a priority that is taken into consideration in the sustainable arrangement of settlements. Criteria for the comfort of living will be the standard of success in evaluating the results of structuring and improving the quality of slums. The success of the physical arrangement of settlements is not a benchmark for the usefulness and sustainability of an area, but a process of survival that will have a positive impact on changes in the quality of life for people who live in Kampung Seng Area Kerantil Settlement

EFEKTIVITAS ANTIFUNGI EKSTRAK ETANOL DAN MINYAK ATSIRI DAUN SIRIH (Piper betle) TERHADAP Trichophyton verrucosum SECARA IN VITRO

Daun sirih (Piper betle L.) diketahui mempunyai efek antifungi. Penelitian ini bertujuan untuk mengetahui efek daun sirih sebagai antifungi secara in vitro. Penelitian dilakukan di Laboratorium Mikologi, Balai Besar Penelitian Veteriner (BB Litvet) tahun 2012. Bahan penelitian adalah ekstrak etanol dan minyak atsiri. Fungi (cendawan) yang digunakan dalam pengujian ini adalah Trichophyton verrucosum, yaitu cendawan jenis kapang, penyebab penyakit kurap pada kulit (dermatofitosis) hewan dan manusia. Media yang digunakan, Sabouraud Dextrose Agar (SDA). Hasil penelitian ditentukan dengan perolehaan nilai konsentrasi hambat minimal (KHM). Bahan uji diencerkan dengan pengenceran ganda 6,250; 3,125; 1,56; 0,78; dan 0,39% untuk ekstrak etanol, dan 12,50; 6,250; 3,125; 1,56; 0,78; dan 0,39% untuk minyak atsiri. Koloni cendawan yang diuji ditumbuhkan pada media SDA dalam cawan petri selama 7 hari, masing-masing disuspensikan dengan aquades steril. Suspensi terdiri dari hifa dan sel spora. Kedua bahan yang diuji dituangkan ke dalam cawan petri steril dengan perbandingan 1:1, masing-masing 1 ml. Suspensi cendawan tanpa ekstrak atau minyak atsiri dituangkan ke dalam cawan petri sebagai kontrol negatif (0%). Media SDA yang masih cair dituangkan sebanyak 20 ml ke masing-masing petri. Inkubasi pada suhu 37oC selama 7 hari. Pengujian dilakukan dua kali ulangan. Hasil penelitian menunjukkan bahwa konsentrasi hambat minimal (KHM) ekstrak etanol adalah 1,56%, dan minyak atsiri 12,5%. Ekstrak etanol mempunyai efek antifungi yang lebih kuat dibandingkan dengan minyak atsiri

REPEATED FREEZE-THAW CYCLES BUT NOT SHORT-TERM STORAGE OF FECAL EXTRACTS AT AMBIENT TEMPERATURE INFLUENCE THE STABILITY OF STEROID METABOLITE LEVELS IN CRESTED MACAQUES

The objective of this study was to examine the effects of repeated freeze-thaw cycles and short-term storage of fecal extracts at ambient temperature on the stability of fecal glucocorticoid (fGCM) and estrogen metabolite (fEM) levels from crested macaques.In total 100 aliquots of fecal extracts from fecal samples collected from female crested macaques (Macaca nigra) living at the Tangkoko-Batuangus Nature Reserve, North Sulawesi were used. We performed two different experiments: (1) An experiment to investigate if levels of fGCM and fEM measured from fecal extracts that were exposed to two, four, six and eight repeated freeze-thaw cycles (test groups) differ to control samples (i.e. fecal extracts always stored frozen); (2) An experiment to evaluate whether storing fecal extracts at ambient temperature for two, four, six, and eight days (test groups)affects the levels of fGCM and fEM compared to the control group (i.e. fecal extracts frozen immediately).Results showed that hormone levels were significantly increased (P0.05) after four freeze-thaw cycles for fGCM and after eight freeze-thaw cycles for fEM. By contrast, there was no significant difference (P0.05) in levels of fGCM and fEM between the test groups and the control group in fecal extracts stored at ambient temperature. In conclusion, our data show that more than two and six repeated freeze-thaw cycles should be avoided when measuring fGCM and fEM in crested macaque fecal extracts, respectively. We also demonstrate that storing fecal extracts at ambient temperature is possible for at least 8 days without taking a risk of affecting the stability of fGCM and fEM levels

Pengembangan metode analisis reserpina secara spektrofotometri dengan pembentukan kompleks perpindahan muatan = Development of spectrophotometric assay method for reserpine based on formation of a charge transfer complex

ABSTRACT: Telah dilakukan penelitian pengembangan metode penetapan kadar reserpina secara spektrofotometri dengan cara pembentukan kompleks perpindahan muatan dan perbandingan metode yang dikembangkan dengan metode penetapan kadar reserpina menurut Farmakope Indonesia III (secara titrasi bebas air). Dad data penelitian yang o\u27iperoleh dilakukan analisis statistik yang meliputi uji ketelitian, ketepatan, serta perbedaan antara kedua metode. Reserpina dapat ditetapkan kadarnya setelah dibentuk menjadi senyawa kompleks perpindahan muatan dengan larutan iodium. Kornpleks yang terjadi mempunyai puncak serapan pada 354 nm, waktu operasional menit ke 9 sampai menit ke 25 setelah reaksi sempurna. Hubungan antara serapan dan kadar merupakan hubungan yang linier dengan batas\u27deteksi 1,25 x 1D-5 M. Hasil perbandingan metode spektrofotometri perpindahan muatan dengan .metode baku Farmakope Indonesia III menunjukkan bahwa kedua metode tidak memberikan hasil analisis yang berbeda secara bermakna. Ketelitian kedua metode juga tidak berbeda secara bermakna, sedang ketepatannya cukup tinggi terbukti dengan harga koefisien variasi yang kecil. Kata kunci: Reserpina, iodium, kompleks perpindahan muatan, spektrofotometr

STEROID LEVEL AND PREGNANCY RATE OF ACEH COWS IN RESPONSE TO OVULATION INDUCTION USING PRESYNCHOVSYNCH METHOD

Penelitian ini bertujuan mengetahui peningkatan level steroid dan persentase kebuntingan sapi aceh terhadap induksi ovulasi dengan metode presynch-ovsynch. Dalam penelitian ini digunakan sepuluh ekor sapi aceh betina dengan status tidak bunting, minimal dua bulan pascapartus, sudah pernah beranak, dan sehat secara klinis. Sapi dibagi atas dua kelompok, yang masing-masing terdiri atas lima ekor sapi. Kelompok pertama (K1) disinkronisasi berahi dengan metode presynch-ovsynch. Pada kelompok kedua (K2), disinkronisasi berahi menggunakan 5 ml PGF2α secara intramuskulus dengan pola penyuntikan ganda dengan interval 12 hari. Setelah  48 jam akhir perlakuan, sapi pada K1 dan K2 diinseminasi menggunakan semen beku fertil. Observasi berahi dilakukan setelah penyuntikan terakhir. Koleksi darah untuk pemeriksaan level estradiol dilakukan segera setelah inseminasi dilakukan sedangkan koleksi darah untuk pemeriksaan progesteron dilakukan pada hari ke-7 pasca-inseminasi. Level steroid diukur menggunakan teknik enzyme-linked immunosorbent assay (ELISA). Pemeriksaan kebuntingan dilakukan 90 hari pasca-inseminasi menggunakan teknik palpasi rektal. Seluruh sapi menunjukkan gejala berahi setelah perlakuan. Level estradiol dan progesteron pada K1 vs K2 masing-masing adalah 294,98±110,48 vs 392,76±11,6 pg/ml (P0,05) dan 23,85±15,14vs 12,69±5,64ng/ml (P0,05). Persentase kebuntingan pada K1 vs K2 masing-masing adalah 60,0 vs 0,0%. Dari hasil penelitian disimpulkan bahwa metode presynch-ovsynch tidak dapat meningkatkan level steroid tetapi dapat meningkatkan persentase kebuntingan pada sapi aceh

The preparation of Dermatophilus congolensis antigen and its testing by means of immunodiffusion test and electrophoresis

The filtrate antigen ofDermatophilus congolensis was prepared based on the Makinde method, whereas the whole cell antigen was based on the Bida and Kelley method. Filtrate antigen of Dermatophilus congolensis has been tested with positive serum from experimental animals, guinea pigs and sheep by means of immurrodiffitsion test and electrophoresis . Positive serum was produced by inoculation of whole cell antigen of D. congolensis to the animals . The results showed that the immunodiffusion test resulted in one and two precipitation lines with positive serum of sheep and guinea pigs respectively. Electrophoresis SDS-PAGE presented about 8 bands with molecular weight in the range from above 30 kD to more than 94 kD. The bands were then transferred into nitrocellulose membrane and gave positive reaction with positive serum from sheep.   Key words : Antigen, Dermatophilus congolensis, immunodiffusion, electrophoresi