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All Journal Buletin Veteriner Udayana
I Gusti Ngurah Kade Mahardika
Laboratorium Virologi Veteriner, Fakultas Kedokteran Hewan Universitas Udayana, Jl. PB. Sudirman, Sanglah, Denpasar, Bali; Indonesia, 80234
Immunology & microbiology Public Health Veterinary

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CASE STUDY OF NEWCASTLE DISEASE VELOGENIC STRAIN WITH MULTIORGAN LESSIONS IN VACCINATED BROILER IN SUSUT, BANGLI I Gde Andhika Putra Pratama; I Gusti Ngurah Kade Mahardika; Ida Bagus Oka Winaya; I Nengah Kerta Besung; Ida Bagus Made Oka
Buletin Veteriner Udayana Bul. Vet. Udayana. June 2025 Vol. 17 No. 3
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i03.p16

Abstract

Newcastle disease (ND) is a highly contagious viral disease of poultry with high mortality rates in velogenic strains. This paper present findings about velogenic strain of Newcastle disease in case study. The methods used in examination of animal case are observation, anatomical pathology examination, histopathology examination, TAB cultivation, serological tests, bacterial identification and parasite identification. In this case study, 28-day-old broiler chickens were obtained from a farmer in Demulih Village, Susut, Bangli. The clinical symptoms observed in the affected chickens included weakness, anorexia, difficulty breathing, greenish-white diarrhea, and torticollis. Pathological anatomical examination revealed congestion in the brain, hemorrhages in the trachea, lungs, heart, and intestines, as well as petechiae in the proventriculus. Histopathological examination showed inflammatory changes were dominated marked by mononuclear cell infiltration in the brain, trachea, lung, heart, proventriculus and intestine. Result of TAB cultivation continued with using HA test showed that virus can agglutination erythrocytes with titer 210 HA Unit. ND virus was confirmed by HI test. Based on this case study, it was concluded that the chickens were infected with ND.
SECONDARY ANTIBODY TITER RESPONSE IN PIGS FOLLOWING STREPTOCOCCUS SUIS VACCINATION WITH VARIOUS ADJUVANTS: DETECTION USING SUPERNATANT ANTIGEN-BASED ELISA Gede Gita Pratama; I Nengah Kerta Besung; Hapsari Mahatmi; I Gusti Ngurah Kade Mahardika; Ni Ketut Suwiti
Buletin Veteriner Udayana Bul. Vet. Udayana. June 2025 Vol. 17 No. 3
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i03.p09

Abstract

Streptococcus suis is a pathogenic bacterium that causes severe disease in pigs and poses a zoonotic risk to humans. Vaccination is one of the primary preventive strategies, and its effectiveness can be enhanced through the use of appropriate adjuvants. This study aimed to evaluate the secondary humoral immune response in pigs vaccinated with a locally derived S. suis vaccine formulated with two different adjuvants: Montanide ISA 201 VG and Montanide Gel-01. In addition, the study observed the dynamics of antibody titters over time post-vaccination. Twelve pigs were divided into three treatment groups: a control group (unvaccinated), a group vaccinated with Montanide ISA 201 VG, and a group vaccinated with Montanide Gel-01. Blood samples were collected weekly for five weeks following the booster vaccination, and antibody titers were measured using ELISA based on culture supernatant antigens. Data were analyzed using analysis of variance and linear regression. The results showed that both adjuvants significantly increased antibody titers compared to the control (p < 0.05), with Montanide Gel-01 producing the highest titer (0.718 ± 0.0397), followed by ISA 201 VG (0.703 ± 0.0320), and the control group (0.459 ± 0.0419). Furthermore, the timing of serum collection significantly influenced antibody titer levels (p < 0.005). This study concludes that the use of Montanide adjuvants, particularly Gel-01, is effective in enhancing the humoral immune response to S. suis vaccination. Further research is recommended to evaluate the effects of increasing antigen concentration for achieving a more robust immune response.
OPTIMIZATION OF ISOPROPYL--D-THIOGALACTOPYRANOSIDE (IPTG) CONCENTRATION AS A TRIGGER FOR BOVINE LACTOFERRINE (BLF) GEN EXPRESSION Luh Dewi Anggreni; Ni made Ritha Krisna Dewi; I Gusti Ngurah Kade Mahardika; I Gusti Ngurah Narendra Putra
Buletin Veteriner Udayana Bul. Vet. Udayana. December 2024 Vol. 16 No. 6
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2024.v16.i06.p22

Abstract

Lactoferrin in cow's milk is known as Bovine Lactoferrin (bLf). Currently, lactoferrin is being developed as an ingredient in vaccines and medicines. As a vaccine material, the plasmid gene bLf is expressed with Isopropyl-b-D-Thiogalactopyranoside (IPTG) to form recombinant proteins. The volume of IPTG used to induce gene expression depends on the bacteria's volume. Different concentrations of IPTG can generally affect the expression rate of recombinant proteins. Therefore, this study determined the optimal concentration of IPTG in expressing bLf protein. The study began with a gradual bLf culture from 1ml of Terrific Broth (TB) media, if the growth is good, upscaling to 10ml culture media. The bLf culture was incubated in a shaker incubator at a speed of 120rpm for 48 hours, at a temperature of 37ºC. After the growth of good bacteria, it was induced using IPTG with varying final concentrations of 0.125mM, 0.25mM, 0.5mM and 1mM. Re-incubated in a shaker incubator for 4-6 hours. The bLf culture was then centrifuged for 10 min at 6000rpm. Centrifugation results in the form of supernatants were removed, while the sediment was added as much as 300μl of PBS. The culture was ultrasonicated for 10 years with an amplitude of 50%. Furthermore, it was analyzed using the SDS-PAGE method. The test began by adding a buffer of 15μl to 45μl of sonicated results and heated at 95ºC for 10 minutes, then electrophoresis on acrylamide gel. The results showed that in each treatment or replicate there was a bLf protein. Visually, it was seen that there was no significant difference in the variation of IPTG concentration treatment for expressing the bLf plasmid gene. Adding IPTG to express bLf genes with lower or higher concentrations did not affect the bLf proteins produced. Lowering the IPTG concentration from 1μM to 0.125 can reduce production costs. Further testing is needed to use lower concentrations of IPTG in other plasmid genes.
THE EFFECTS OF TRANSFORMATION DURATION ON THE QUANTITY OF SWINE PATHOGENIC ESCHERICHIA COLI ISOLATE Kalyana Lionita; I Gusti Ngurah Kade Mahardika; Anak Agung Sagung Kendran
Buletin Veteriner Udayana Bul. Vet. Udayana. April 2025 Vol. 17 No. 2
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i02.p04

Abstract

DNA transformation generally still utilizes commercial Escherichia coli bacteria, and the use of virulent pathogenic bacteria has never been reported. Utilizing virulent pathogenic bacteria can provide an advantage, as the product can be used as a vaccine against the genetic material in the plasmid and against the pathogenic bacteria itself. This study aims to determine how the transformation duration affects the number of swine Escherichia coli pathogenic isolates colonies. The samples of this study are swine Escherichia coli pathogenic isolates BPOS4 and E. coli BL21 (DE3) which were made into competent cells by calcium chloride heat shock method and then transformed with the different transformation of times using plasmid pGEX-2T+ASF A224L. The results showed that E. coli BL21 (DE3) and swine Escherichia coli pathogenic isolates BPOS4 had no significant effect on the quantity of colonies after transformation. Although statistically the effect was not significant, E. coli BL21(DE3) gave a higher colony count with an average of 62,4×105 CFU/100µL compared to E. coli BPOS4 of 33,45×105 CFU/100µL. The transformation time has no significant effect on the number of bacterial colonies after transformation, however, the best transformation time is shown by the treatment of 60 seconds with an average colony of 86,5×105 CFU/100µL. It can be concluded that the colonies of E. coli BL21 (DE3) and E. coli BPOS4 transformed with plasmid pGEX-2T+ASF A224L can grow optimally in 60 seconds. Further research can be conducted, such as developing Escherichia coli vaccines using pathogenic strains.
THE IMMUNOLOGICAL RESPONSE OF SWINE TO AFRICAN SWINE FEVER DNA VACCINE INDUCTION Kadek Thalia Suryaningtyas Oka Wardhani; I Gusti Ngurah Kade Mahardika; I Ketut Berata
Buletin Veteriner Udayana Bul. Vet. Udayana. April 2025 Vol. 17 No. 2
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i02.p23

Abstract

African Swine Fever (ASF) is a disease that attack swine with high mortality. DNA vaccine against ASF had been tested in mouse, but the research in swine has not been done. The purpose of this study is to evaluate swine immune response against ASF DNA vaccine and time effect of serum collection against antibodies of ASF virus. The method that had been used is vaccinating the swine with ASF DNA vaccine intramuscularly and measuring humoral immune response with ELISA on collected swine serum in weeks 0, 2, and 4. The result pointing that ASF DNA vaccine can induce production of antibodies in treatment group compared with control group with Optical Density (OD) increasing value in weeks 2 and 4. The serum time collection also do affect antibodies level in every weeks that had been observed, that is weeks 0, 2, and 4 with significant increase between week 0 and 2, but between week 2 and 4 the antibodies level is not affected significantly. This study is recommending further research to observe protection effect of ASF DNA vaccine by extending research duration and observation of celluler immune response to support vaccine effectiveness in field.
DETECTION OF PRIMARY ANTIBODIES AGAINST SUPERNATANT ANTIGENS IN PIGS VACCINATED WITH INACTIVATED STREPTOCOCCUS SUIS USING DIFFERENT ADJUVANTS I Putu Heru Darawinata; I Nengah Kerta Besung; I Gusti Ngurah Kade Mahardika
Buletin Veteriner Udayana Bul. Vet. Udayana. April 2025 Vol. 17 No. 2
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i02.p33

Abstract

Streptococcus suis is a pathogenic bacterium that causes disease in pigs and has zoonotic potential, leading to economic losses in the livestock industry and posing risks to human health. This experimental study employed a factorial completely randomized design to evaluate the primary antibody response to supernatant antigens in pigs vaccinated with inactivated Streptococcus suis vaccines formulated with different adjuvants. Twelve pigs were randomly assigned to three treatment groups: a control group (receiving only adjuvant without antigen), a vaccine group with Montanide ISA 201 VG adjuvant, and a vaccine group with Montanide Gel 01 adjuvant. Serum samples were collected weekly for five weeks, and antibody titers were measured using an enzyme-linked immunosorbent assay (ELISA).The results demonstrated that both adjuvanted vaccines significantly increased antibody titers compared to the control group. Antibody production was detectable from the second week post-vaccination, reaching peak levels in the fourth week. Comparative analysis revealed that Montanide ISA 201 VG induced slightly higher antibody responses than Montanide Gel 01. These findings support the use of inactivated Streptococcus suis vaccines with adjuvants as an effective preventive strategy to enhance immunity in swine populations and reduce the risk of zoonotic transmission.
ANTIBODY RESPONSE IN MICE IMMUNIZED WITH ASF_703 DNA VACCINE Fidella Luthfia Qotrunnada; I Gusti Ngurah Kade Mahardika; Hamong Suharsono
Buletin Veteriner Udayana Bul. Vet. Udayana. August 2025 Vol. 17 No. 4
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i04.p03

Abstract

African Swine Fever (ASF) is a highly contagious viral disease in pigs, causing significant economic losses due to high mortality rate and the lack of an effective vaccine. On going vaccine development efforts include research at the Biomedical Laboratory of the Faculty of Veterinary Medicine, Udayana University, which previously developed the ASF_701 DNA vaccine. The study aimed to evaluate the immune response in mice to a novel ASD DNA vaccine formulation containing four additional genes (EP153R, A238L, DP96R, and S276R) and a polymer adjuvant. In this experimental study, mice were divided into two groups: a control group and a vaccinated group. Each mouse in the vaccinated group received a subcutaneous injection of 25 µg plasmid DNA in polymer adjuvant. The immune response was assessed using ELISA to detect ASF-specific antibodies at weeks 0, 1, 2, 3, and 4, with synthetic peptides as coating antigens. Optical Density (OD) values were statistically analyxed using SPSS version 25. The results demonstrated a significant difference (p < 0.01) in antibody levels between vaccinated and control mice, with serum collection time also showing a significant effect (p = 0.004). The finfings suggest that the new vaccine formulation elicits a measurable immune response. However, further studies are needed to improve the stability and long-term efficacy of the vaccine.
CASE REPORT: COINFECTION OF TRICHURIS SUIS AND STREPTOCOCCUS SP. IN A LANDRACE-YORKSHIRE PIGLET FROM BUAHAN VILLAGE, PAYANGAN, GIANYAR Viviana Anyaputri Tanurahardja; I Ketut Berata; I Nengah Kerta Besung; I Putu Cahyadi Putra; I Gusti Ngurah Kade Mahardika
Buletin Veteriner Udayana Bul. Vet. Udayana. August 2025 Vol. 17 No. 4
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i04.p37

Abstract

Pig farming in Bali plays a strategic role in meeting both animal protein demands and cultural needs, yet remains highly susceptible to viral, bacterial, and parasitic diseases. This case report documents a severe coinfection of Trichuris suis (trichuriasis) and systemic beta-hemolytic Streptococcus sp. (presumably S. suis) in a 3.5-month-old weanling piglet from Gianyar. Diagnostic methods included anamnesis, epidemiological investigation, gross pathology, histopathology, bacterial culture/identification, and parasitic examination. The piglet exhibited stunted growth, cachexia, cough, and chronic brown diarrhea. Gross and histopathological findings revealed meningoencephalitis, necrotic-edematous bronchopneumonia, and edematous-degenerative typhlitis. Bacterial isolation identified beta-hemolytic Streptococcus sp. in the brain and lungs, though neurological signs were absent. Necropsy uncovered ~4,700 T. suis in the cecum and colon, with Entamoeba sp. cysts detected in feces. This case highlights: (1) the clinicopathological manifestations of concurrent T. suis and Streptococcus sp. infections, and (2) the critical need for early detection and comprehensive diagnostics in field cases. To mitigate such coinfections, we recommend enhanced biosecurity, routine antiparasitic treatment, and periodic bacteriological surveillance.
FATAL CANINE PARVOVIRUS INFECTION WITH SECONDARY BACTERIAL AND PARASITIC COMPLICATIONS IN A PUPPY: A CASE REPORT Egrina Sonta Bako; I Putu Cahyadi Putra; Ida Bagus Oka Winaya; I Gusti Ngurah Kade Mahardika; I Nengah Kerta Besung
Buletin Veteriner Udayana Bul. Vet. Udayana. August 2025 Vol. 17 No. 4
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i04.p13

Abstract

Canine parvovirus (CPV) is a highly infectious viral disease that poses a significant threat to puppies, often resulting in fatality if not diagnosed and treated promptly. This report investigates the cause of death of a nearly two-month-old puppy in Denpasar City. Diagnosis was confirmed through a comprehensive approach, including a thorough history, clinical examination, hematology, anatomical pathology, histopathology with hematoxylin and eosin (H&E) staining, polymerase chain reaction (PCR) for CPV, bacteriology, and parasitology examinations. Anamnesis revealed that the two puppies succumbed after exhibiting symptoms such as anorexia, vomiting, bloody diarrhea, dehydration, and cachexia, which were also observed in this puppy. Necropsy findings indicated multiorgan lesions, including hemorrhage, congestion, hyperemia, uneven organ discoloration, swelling of the heart and spleen, and worm infestation in the stomach. Histopathological analysis revealed multi-organ inflammation characterized by hemorrhage, necrosis, edema, and inflammatory cell infiltration in all organs, with lesions typical of CPV, including lymphoid follicular necrosis and intestinal Lieberkühn crypts in the intestine. PCR confirmed CPV infection, as evidenced by a 900 bp DNA band. Bacteriological examination revealed the presence of Escherichia coli and Streptococcus spp. in the intestine and Staphylococcus spp. in the spleen. Parasitological examination revealed the presence of adult worms, larvae, and eggs of Toxocara canis in the fecal sample. Based on these findings, the death of the puppy was attributed to CPV infection compounded by secondary bacterial and parasitic infections. Further testing is recommended to ascertain potential co-infection with Canine Distemper Virus.
ANTIBODY RESPONSE IN MICE TO FOOT AND MOUTH DISEASE (FMD) DNA VACCINE I Gede Wiraguna Arsadita; I Gusti Ngurah Kade Mahardika; Hamong Suharsono
Buletin Veteriner Udayana Bul. Vet. Udayana. August 2025 Vol. 17 No. 4
Publisher : Fakultas Kedokteran Hewan Universitas Udayana

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24843/bulvet.2025.v17.i04.p32

Abstract

Foot and Mouth Disease (FMD) is a highly contagious viral infection in livestock, causing severe economic losses worldwide. While conventional vaccines are widely used, innovative alternative such as DNA vaccines is needed. This study aimed to assess the immunogenicity of a novel DNA vaccine against FMD, encoding the VP0, VP3, VP1, 2A, and 3CPRO genes derived from an Indonesian FMD virus isolate (2022). Female mice were immunized with the vaccine, and antibody responses were evaluated weekly for four weeks using ELISA using synthetic peptide of VP1 as coating antigen. Results demonstrated that the vaccine significantly induced FMD-specific antibodies compared to the control group, with optical density (OD) values consistently higher in vaccinated mice across all time points post vaccination (p < 0.001). However, no significant difference in antibody levels was observed between weeks post-vaccination within the treatment group (p = 0.678), suggesting stable immune activation after the initial response. These findings indicate that the DNA vaccine effectively triggers humoral immunity, supporting its potential as a viable alternative to traditional FMD vaccines. Further research should evaluate its efficacy in target livestock species and optimize delivery methods for field applications.
Co-Authors Anak Agung Sagung Kendran Egrina Sonta Bako Fidella Luthfia Qotrunnada Gede Gita Pratama Hamong Suharsono Hamong Suharsono Hapsari Mahatmi I Gde Andhika Putra Pratama I Gede Wiraguna Arsadita I Gusti Ngurah Narendra Putra I Ketut Berata I Nengah Kerta Besung I Nengah Kerta Besung I Nengah Kerta Besung I Nengah Kerta Besung I Putu Cahyadi Putra I Putu Cahyadi Putra, I Putu Cahyadi I Putu Heru Darawinata Ida Bagus Made Oka Ida Bagus Oka Winaya Kadek Thalia Suryaningtyas Oka Wardhani Kalyana Lionita Luh Dewi Anggreni Ni Ketut Suwiti Ni made Ritha Krisna Dewi Viviana Anyaputri Tanurahardja