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Abstract Brucellosis is a zoonotic disease that cause a significant economic losses for cattle industries worldwide. A rapid, precise and accurate diagnosis technique for diagnosis of brucellosis in all stages of the infection is definitely required. Blood-samples are widely used for PCR-based DNA analysis because they are easily collected, handled, and processed. Direct PCR analysis without DNA extraction has been attempted to reduce time and costs for routine analysis. This approach is promising but is still limited by the presence of PCR inhibitors that is naturally found in the blood samples. The objective of this study was to compare the effectivity of direct PCR technique with or without DNA extraction for detection of Brucella abortus in the blood samples. Three whole-blood samples from brucella infected dairy cattle and five whole-blood samples from beef cattle that having abortion were used as samples in this study. A pair of bcsp31 primers and IS711 primers were used for amplification of genus-specific and species-specific of Brucella. The results showed that amplicon in the position of 223 bp and 498 bp that are specific for B. abortus were detected from all of the samples that were analyzed on 1.5% agarose gels. Based on the result it could be concluded that direct PCR analyses without DNA extraction is a sensitive, specific, simple, rapid and inexpensive assay for detecting B. abortus in the whole blood samples for either dairy or beef cattle and therefore it could improve the existing surveillance and control programs for brucellosis. Keywords : brucellosis; direct PCR; PCR inhibitor; whole-blood sample; without DNA extraction Abstrak Brucellosis adalah penyakit zoonosis yang menyebabkan kerugian ekonomi yang signifikan bagi industri ternak di seluruh dunia. Teknik diagnosis yang cepat, tepat dan akurat yang dapat digunakan untuk diagnosis brucellosis pada semua tahap infeksi sangat diperlukan. Sampel darah banyak digunakan untuk analisis PCR berbasis DNA karena mudah untuk dikoleksi, ditangani, dan diproses. Metoda PCR langsung tanpa didahului dengan ekstraksi DNA dikembangkan dengan tujuan penghematan waktu dan beaya untuk analisa secara rutin. Tehnik ini sangat menjanjikan tetapi memiliki keterbatasan karena adanya senyawa penghambat PCR yang secara alami terkandung di dalam sampel darah . Tujuan dari penelitian ini adalah membandingkan efektifitas antara uji PCR secara langsung dengan ekstraksi dan tanpa ekstraksi DNA untuk deteksi Brucella abortus di dalam darah. Tiga ( 3 ) sampel darah-EDTA yang berasal dari sapi penderita brucellosis dan 5 sampel darah-EDTA dari sapi potong yang mengalami abortus digunakan sebagai sampel dalam penelitian ini. Pasangan primer bcsp31 dan primer IS711 untuk amplifikasi gen dan species specific digunakan dalam penelitian. Hasil menunjukkan bahwa amplikon/pita pada posisi 223 bp dan 498 bp yang spesifik untuk Brucella abortus terdeteksi dari semua sampel yang dianalisa dengan gel agarosa 1,5%. Berdasarkan hasil penelitian dapat disimpulkan bahwa uji PCR secara langsung tanpa didahului dengan ekstraksi DNA merupakan tehnik yang sensitif, spesifik, sederhana, cepat dan murah untuk deteksi B. abortus di dalam sampel darah baik sapi perah maupun sapi potong dan oleh karena itu diharapkan dapat digunakan untuk memperbaiki program kontrol dan survailance yang telah ada untuk brucellosis. Kata kunci : brucellosis; PCR langsung; penghambat PCR; sampel darah-utuh; tanpa ekstraksi DNA

Leptospirosis is a zoonotic disease of global concern, and is caused by pathogenic serovar Leptospira interrogans. Canine Leptospirososis is widespread worldwide, dogs can act as incidental hosts or maintenance hosts for various serovars. The purpose of this research was to identify leptospire serovars that infect healthy and suspected leptospirosis dogs in Yogyakarta. A total of 56 dogs (36 healthy dogs and 20 suspect leptospirosis dogs) sera were taken from cephalica vein as much as 3 ml. Sera were examined for leptospirosis with Microscopic Aglutination Test (MAT) which conducted at the Research Center for Veterinary Science, Bogor. Microscopic Aglutination Test carried out on various Leptospire serovar, namely: Ichterohaemorrhagiae, Javanica, Celledoni, Ballum, Pyogenes, Cynopeteri, Rachmati, Australis, Pomona, Canicola, Grippotyphosa, Bataviae, Hardjo, and Tarrasovi. The results showed that Celledoni serovars infected 25% of healthy dogs and 5% of suspect leptospirosis dogs, Javanica serovar infected 19% of healthy dogs, Bataviae serovars infected 15% of suspect leptospirosis dogs, Grippotyphosa serovar infected 11% of healthy dogs, Tarrasovi serovar infected 10% of suspect leptospirosis dogs, serovars Cynopteri infects 5% of healthy dogs and 5% of suspect leptospirosis dogs, serovar Pyrogenes infects 5% of healthy dogs and 5% of suspect leptospirosis dogs, and serovar Rachmati infects 5% of suspect leptospirosis dogs. Seven healthy dogs (19%) and 2 suspect leptospirosis dogs (10%) were infected with more than 2 leptospire serovars. From the results of this study it can be concluded that Celledoni serovar of Leptospira interrogans infection causes subclinical leptospirosis, while Bataviae serovar infection causes clinical leptospirosis in dogs in Yogyakarta.

Poultry can be infected not only by one type of virus, but also by more than one type of virus. Avian influenza virus (AIV) and Newcastle disease virus (NDV) are two important pathogens in poultry that can cause recurrent outbreaks and often lead to clinical symptoms and pathological lesions similar in poultry with high morbidity and mortality causing significant economic losses to the poultry industry. This study aims to detect and differentiate pathogenic viruses in commercial laying chickens with similar clinical symptoms, especially torticolis and curled toe paralysis, and anatomic pathological lesions, such as foci necrotic hemorrhages in the digestive tract by applying streptavidin-biotin immunohistochemical immunological test (IHC SB). Samples (chickens) were taken from the cases of the disease in poultry in several commercial poultry farms. The layer chickens in which showed clinical symptoms of torticolis and curled toe paralysis, and hemorrhagic lesions in the lungs and / or digestive system are suspected of having AIV and NDV infections. After being necropsied, then lungs and gastrointestinal tract were tested IHC SB. The IHC SB results were examined with a digital microscope camera system. The results of the present study indicated that AIV antigen was detected in the lungs, but not in the digestive tract, whereas, NDV was detected in the digestive tract, but not in the lungs. It was concluded, that the IHC SB can be applied for confirmation of the diagnosis of AIV and NDV and indicated, that the commercial laying chickens with clinical symptoms of torticolis and curled toe paralysis, as well as pathological hemorrhagic lesions in the gastrointestinal tract could be infected by both viruses.

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The use of herbal medicine (phytopharmaca) has increased manifold because of the safety. Red fruit extract is one of the supplements that has many advantages for health. The efficacy of pre-clinical trial using the rats induced by iodine deficient ration has proven that 1-2 ml of red fruit extract supplementation with/without KIO3 fortification for 30, 60 and 90 days has potency as anti-goitrogenic herbal medicine. For standarization, however, the extract red fruit has to be tested for the safety in the important organs like livers and kidneys. The research results have proven that in the normal Sprague Dawley rats, supplementation of 2 ml/day/rat of red fruit extract for 135 consecutive days has.no toxic effect in the livers and kidneys. In hyperplastic goiter Sprague Dawley rats induced by goitrogenic ration, the red fruit extract supplementation has to be in a combination with KIO3 to overcome iodine deficiency disorders and has no-toxic effect to the livers. Therefore, it can be concluded that based on pre-clinical studies using Sprague Dawley rats, the red fruit extract can be categorized as standardized anti-goitrogenic herbal medicine.