Search

Background and Aim: Bovine viral diarrhea (BVD), a highly pathogenic ribonucleic acid (RNA) virus, causes devastating financial losses and reproductive deaths among dairy cattle in Yogyakarta and globally. This study aimed to identify point mutations within the E2 structural protein of the acquired BVD virus (BVDV) isolates using genetic analysis. Materials and Methods: The study period shows that we performed the research in 2023. We collected 118 serum samples from 2019 to 2023, among which only 10 BVDV positive were used and 108 were negative lacking the BVDV antigen. An anti-Erns monoclonal antibody-coated protein was used in indirect antigen capture enzyme-linked immunosorbent assay (I-ACE) to detect the BVD antigen present in positive BVDV serum specimens. In the initial step of the two-step reverse transcription polymerase chain reaction, the enzyme (superscript III reverse transcriptase) and the primer (random hexamer) were used to convert the RNA of the BVDV into complementary deoxyribonucleic acid (cDNA) during the process of reverse transcription. The final step involved the amplification of the E2 gene of the resultant BVDV cDNA through gene-specific primers (E2_fwd: 5′-TGGTGGCCTTATGAGAC-3′ and P7_rev: 5′-CCCATCATCACTATTTCACC-3′) and enzyme (platinum taq DNA polymerase high fidelity). For conducting Sanger sequencing, those 3 BVDV-1-positive isolates (about 2.6% of all isolates) were selected as a typical specimen for each site and year between 2019 and 2023 using a proportional computation. Therefore, only two BVDV isolates with complete genomes were chosen to perform their homological and genetic analysis based on the E2 gene by means of Blast and MEGA Version 11 in addition to the Bioedit 7.2.5 program. Results: By applying phylogenetic analysis relying on the E2 gene, a sum of 1011 nucleotides of the BVDV-1 isolates derived from each of the two BVDV-1 Indonesian isolates (n = 2) and its 23 reference BVDV strains were acquired from the National Center for Biotechnology Information (NCBI) database. The findings of the genetic analysis inside the phylogenetic tree revealed that the two BVDV Indonesian isolates were clustered into BVDV-1a subgenotype, while the reference BVDV strains were clustered into the five BVDV subgenotype, BVDV-1a (n = 6), BVDV-1b (n = 3), BVDV-1c (n = 11), BVDV-1m (n = 1), and BVDV-1n (n = 2). The branch exists in phylogenetic tree located before the division of our two BVDV isolates was divided into two branches with the same maximum bootstrap values of 99%, indicating a high degree of confidence, was seen. Next, we observed the branch near our study samples, which displayed the bootstrap value of 100, indicating that our 02 isolates were identical. In both isolates, V11 BVDV1/Indonesia/Yogyakarta/2023 and V16 BVDV1/Indonesia/ Yogyakarta/2023 with GenBank accession numbers PP836388 and PP836389, respectively, conserved D7E residues were mutated as well as cysteine changed/altered into serine (S) was identified at amino acid position 201. Conclusion: We identified two isolates of BVDV belonging to the BVDV-1a subgenotype. Our findings indicate that the conserved D7E residues of isolates V11 BVDV1/Indonesia/Yogyakarta/2023 and V16 BVDV1/Indonesia/Yogyakarta/2023 were altered. The Indonesian BVDV isolates exhibited a cysteine to serine mutation at amino acid position 201, leads to vaccination failure, range of animal’s host will increase, and diagnostic kit will not be effective.

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

Abstract Bovine Viral Diarrhea (BVD) is one of the main causes of impaired productivity and reproduction of cows. Antigen capture Elisa (ACE) is one of the serological technique that is sensitive, reliable and used regularly for detecting persistent BVD infection individually which simpler than multiplex nested PCR. The aim of this study was to determine the agreement between ACE and multiplex nested PCR as a routine laboratory diagnostic technique to detect the presence of BVD infection. A total of 128 cow serum samples consisting of 63 positive and 65 negative samples based on ACE were used in this study. The samples were collected from active and passive surveillance in dairy and beef cattle conducted by Balai Besar Veteriner (BBVet) Wates. The serum samples were then tested molecularly using multiplex nested PCR against BVD. The result showed 48 out of 63 BVDV-1 positive samples were found positive BVD antigen whereas 57 of 65 BVDV-1 negative samples were negative using multiplex nested PCR, . The agreement value between the two different assays based on statistic analysis using Kappa method was 0.64 and classified a good one. The result concluded that the ACE BVD assay was equally suitable as routine diagnosis to determine BVD infected cattle in the farm. Keywords: Antigen capture ELISA; Bovine viral diarrhea; Kappa; Multiplex nested PCR. Abstrak Bovine Viral Diarrhea (BVD) merupakan salah satu penyebab gangguan produktivitas dan reproduksi sapi. Antigen capture ELISA (ACE) merupakan salah satu teknik serologis yang sensitif, dapat diandalkan dan digunakan secara teratur untuk mendeteksi infeksi BVD persisten secara individual yang lebih sederhana daripada multiplex nested PCR. Tujuan penelitian ini adalah untuk mengetahui kesesuaian antara uji ACE dan multiplex nested PCR sebagai teknik diagnostik laboratorium rutin untuk mendeteksi adanya infeksi BVD. Sebanyak 128 sampel serum sapi yang terdiri dari 63 sampel positif dan 65 negatif berdasarkan ACE BVDV Antigen Test Kit/Serum Plus (Idexx®) digunakan dalam kajian ini. Sampel serum sapi merupakan koleksi dari surveilans aktif dan pasif pada sapi perah dan potong yang dilakukan Balai Besar Veteriner (BBVet) Wates. Sampel serum kemudian diuji secara molekuler menggunakan multiplex nested PCR terhadap BVD. Hasil penelitian menunjukkan bahwa dengan teknik multiplex nested PCR, 48 dari 63 sampel positif BVDV-1 ditemukan positif untuk antigen BVD sedangkan 57 dari 65 sampel negatif BVDV-1 negatif untuk antigen BVD. Analisis statitik berdasarkan perhitungan metoda Kappa menunjukkan nilai kesesuaian antara dua uji sebesar 0,64 dan tergolong bagus. Hasil penelitian menunjukkan kesimpulan bahwa uji ACE BVD sesuai sebagai diagnosis rutin untuk menentukan ternak yang terinfeksi BVD di peternakan. Kata kunci: Antigen capture ELISA; Bovine viral diarrhea; Kappa; Multiplex nested PCR.

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.

Avian influenza is a viral disease in poultry caused by avian influenza virus (AIV) subtype H5N1 with varying clinical signs are often similar to the clinical signs of other viral infections, such as Newcastle disease virus (NDV). The mechanism of disease pathogenesis to express clinical signs tightly correlated to the determination of the target organ seen from AIV H5N1 antigens distribution in organs, such as respiratory tract, brain and gastrointestinal tract. Immunopathological immunohistochemistry streptavidin-biotin (IHC SB) is a method for sensitive and accurate in detecting antigens of AIV on the tissues. In the present study, it was determined whether in laying hens with clinical signs of torticollis and curled toe paralysis, and pathologic anatomic lesions in the form of petechial and foci necrotic hemorrhages tested with immunopathological IHC SB is positive AIV H5N1 infection. IHC SB study results showed that the AIV H5N1 antigen were found in tissues of the lung, brain, duodenum and proventriculus. Based on these results, we can conclude that the IHC SB is a method that is highly sensitive and accurate to detect H5N1 antigens and its distribution in the host.

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.

House flies (Musca domestica L.) are one of the major pests found in poultry farms resulting in not onlyannoyance and indirect damage to the poultry production but also transmitting many infectious organismsmechanically and biologically. A highly pathogenic avian influenza virus H5N1 (HPAIV H5N1) collectedfrom field house flies in Java Island, have fully succeeded identified and isolated in 2008. The AIV H5N1isolates were further used in the present study to determine the presence and persistence of the AIV H5N1in the laboratory infected house flies. One hundred house flies from a free AIV poultry farm in Yogyakarta,Indonesia were used in this study. The collected house flies were fasted for 12 hours then divided equallyin control and treated groups. The treated group was allowed to imbibe Dulbecco’s modified eagle medium(DMEM) containing AIV H5N1 while the control group imbibed medium without virus for one hour. Theflies from each group were collected at 12 and 24 hours post-exposure, respectively. All flies were immobilizedat 4°C, immersed in absolute ethanol for a few seconds and dissected under the stereomicroscope to collectthe gastrointestinal (GI) tracts. Based on the RT-PCR results, it is concluded that AIV H5N1 remainedexist in GI tracts of house flies for at least 24 hours post-exposure.

Avian influenza virus subtype H5N1 (AIV H5N1) is highly pathogenic and fatal in poultry. The virusis still endemic with low virulence rate, although it may play a critical role in causing high morbidity andmortality rates in poultry in Indonesia. In general, diagnostic approach for AIV H5N1 is based onconventional serological and viral isolation methods that have the potential to produce consumings oftime and relatively expensive cost within the laboratory without compromising test utility. Thus, amolecular approach of multiplex reverse transcription-polymerase chain reaction (mRT-PCR) was developedand applied for the detection of matrix gene type A influenza viruses, AIV subtype subtype H5hemagglutinin gene with simultaneous detection of N1 nucleoprotein gene. Thirty sera specimens fromthe diseased commercial chickens that were specifically amplified positive-RT-PCR for AIV H5N1 wereselected for mRT-PCR. The mRT-PCR products were visualized by agarose gel electrophoresis and consistedof DNA fragments of AIV of 245 bp, 545 bp and 343 bp for M, H5 and N1 genes, respectively. Thus, themRT-PCR that can rapidly differentiate simultaneously between these genes is very important for thecontrol and even eradication of AIV transmission in poultry in Indonesia.

Calcitriol supplementation in ovariohisterectomized rat was known to decreased calcium retention.The objective of the research was to study the response of femur bone to calcitriol supplementation for 30weeks in ovariohisterectomized rats consuming casein. Twenty female Wistar rats at 8 weeks of age wererandomly divided into four groups (unovariohisterectomy without calcitriol supplementation (N),unovariohisterectomy with calcitriol supplementation (ND), ovariohisterectomy without calcitriolsupplementation (O) and ovariohisterectomy with calcitriol supplementation (OD) of five each. Thirtyweeks after surgery, femur was taken for histopathological and immunohistochemistry examination.Immunohistochemistry of distal femur metaphysis in group O and OD were revealed decreasing tartrateresistant alkaline phosphatase 5b (TRAP5b) in trabecular bone, which was located in bone marrow space,and also in trabecular speculum surface. Histopathological analysis of distal femur metaphysis in groupN and ND were showed normal structure, meanwhile, distal femur metaphysis in group O and OD wereshown some abonormalities, such as increased of bone marrow space, domination of adipocytes in the bonemarrow, and decreased of trabecular bone speculum in metaphysis. Based on the results, it was concludedthat femur bone of ovariohisterectomized rats fed casein with and without calcitriol 8ng/daysupplementation for 30 weeks were showed unbalanced between resorption and formation of bone whichwas domination by bone resorption.

Background The decline in estrogens could lead to an imbalance of bone formation and absorption as well as bone mineralization. Phytoestrogens are known to have the potential to maintain bone health. Calcium and phosphorus are essential components for bone mineralization. This study aimed to investigate the effect of soybean sprout flour consumption on serum calcium and phosphorus levels in ovariectomized rats. Methods A randomized post-test-only control group design involving 25 female Sprague-Dawley rats aged 2 months, which were divided into five groups: (i) normal controls on standard feed diet AIN-93 (C-N), (ii) ovariectomy controls on standard feed diet (OVX -N); (iii) ovariectomized rats on standard feed diet + ethinylestradiol (OVX -E), (iv) ovariectomized rats on standard feed diet + soybean flour (OVX -S); (v) ovariectomized rats on standard feed diet + soybean sprout flour (OVX -SS). The rats were treated for 6 weeks, then on the last day of treatment blood sampling was conducted. Analysis of calcium and phosphorus in serum was performed using the Arsenazo III photometric method. Data were analyzed using one-way Anova. Results Mean serum calcium level was highest in the OVX-SS group (12.59 mg/dL ± 1.32), while mean serum phosphorus level was highest in the C-N group (1.708 mg/dL ± 0.28). Soybean sprout flour consumption had a significant effect on serum calcium levels (p=0.019), yet it had no effect on body weight and serum phosphorus levels of the rats. Conclusion Consumption of soybean sprout flour could positively affect serum calcium levels in rats.

File lengkap ada dalam bentuk PDF dibawah ini

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.

The objective of this study was to analyze the influence of chitosan on coronary artery wall thicknessand lumen diameter of Sprague Dawley rats induced by high fat ration. The animals for this study weretwenty adult male rats divided into four groups, i.e. group I as the control was fed with basal rationcontaining normal fat for 3 months, group II was fed ration containing high fat for three months, group IIIwas fed ration containing high fat and given chitosan 180 mg per kg body weight per day orally in 2 mLaquadest for 3 months, group IV was fed ration containing high fat for three months and after one monthgiven chitosan 180 mg per kg body weight per day orally in 2 mL aquadest for two months. Each groupconsisted of five animals. After 90 days, the rats were necropsied and the hearts were collected tohistopathological. The difference between treatments was statistical analysis by Anava, and continued byLowest Significant Difference Test to locate the difference. Normal fat ration did not influence coronaryartery wall thickness and coronary artery lumen diameter in rats. High fat ration increased coronaryartery wall thickness and chitosan could be decreased coronary artery wall thickness in rats. High fatration + chitosan that given simultaneously could be increased coronary artery lumen diameter in rats.Based on the result of this study, it was concluded that high fat ration was a major factor able to causeatherosclerosis and chitosan was able to prevent atheroma plaque formation.

Background and Aim: Bovine viral diarrhea (BVD), a highly pathogenic ribonucleic acid (RNA) virus, causes devastating financial losses and reproductive deaths among dairy cattle in Yogyakarta and globally. This study aimed to identify point mutations within the E2 structural protein of the acquired BVD virus (BVDV) isolates using genetic analysis. Materials and Methods: The study period shows that we performed the research in 2023. We collected 118 serum samples from 2019 to 2023, among which only 10 BVDV positive were used and 108 were negative lacking the BVDV antigen. An anti-Erns monoclonal antibody-coated protein was used in indirect antigen capture enzyme-linked immunosorbent assay (I-ACE) to detect the BVD antigen present in positive BVDV serum specimens. In the initial step of the two-step reverse transcription polymerase chain reaction, the enzyme (superscript III reverse transcriptase) and the primer (random hexamer) were used to convert the RNA of the BVDV into complementary deoxyribonucleic acid (cDNA) during the process of reverse transcription. The final step involved the amplification of the E2 gene of the resultant BVDV cDNA through gene-specific primers (E2_fwd: 5'-TGGTGGCCTTATGAGAC-3' and P7_rev: 5'-CCCATCATCACTATTTCACC-3') and enzyme (platinum taq DNA polymerase high fidelity). For conducting Sanger sequencing, those 3 BVDV-1-positive isolates (about 2.6% of all isolates) were selected as a typical specimen for each site and year between 2019 and 2023 using a proportional computation. Therefore, only two BVDV isolates with complete genomes were chosen to perform their homological and genetic analysis based on the E2 gene by means of Blast and MEGA Version 11 in addition to the Bioedit 7.2.5 program. Results: By applying phylogenetic analysis relying on the E2 gene, a sum of 1011 nucleotides of the BVDV-1 isolates derived from each of the two BVDV-1 Indonesian isolates (n = 2) and its 23 reference BVDV strains were acquired from the National Center for Biotechnology Information (NCBI) database. The findings of the genetic analysis inside the phylogenetic tree revealed that the two BVDV Indonesian isolates were clustered into BVDV-1a subgenotype, while the reference BVDV strains were clustered into the five BVDV subgenotype, BVDV-1a (n = 6), BVDV-1b (n = 3), BVDV-1c (n = 11), BVDV-1m (n = 1), and BVDV-1n (n = 2). The branch exists in phylogenetic tree located before the division of our two BVDV isolates was divided into two branches with the same maximum bootstrap values of 99%, indicating a high degree of confidence, was seen. Next, we observed the branch near our study samples, which displayed the bootstrap value of 100, indicating that our 02 isolates were identical. In both isolates, V11 BVDV1/Indonesia/Yogyakarta/2023 and V16 BVDV1/Indonesia/ Yogyakarta/2023 with GenBank accession numbers PP836388 and PP836389, respectively, conserved D7E residues were mutated as well as cysteine changed/altered into serine (S) was identified at amino acid position 201. Conclusion: We identified two isolates of BVDV belonging to the BVDV-1a subgenotype. Our findings indicate that the conserved D7E residues of isolates V11 BVDV1/Indonesia/Yogyakarta/2023 and V16 BVDV1/Indonesia/Yogyakarta/2023 were altered. The Indonesian BVDV isolates exhibited a cysteine to serine mutation at amino acid position 201, leads to vaccination failure, range of animal's host will increase, and diagnostic kit will not be effective. [ABSTRACT FROM AUTHOR]