Phylogenetic tree analysis for Bali Cattle based on partial sequence 16S rRNA Mitochondrial DNA

Mitochondria DNA (mtDNA) as a source of genetic information based on the maternal genome, can provide important information for phylogenetic analysis and evolutionary biology. The objective of this study was to analyze the phylogenetic tree of Bali cattle with seven gene bank references (Bos indicus, Bos taurus, Bos frontalis, and Bos grunniens) based on partial sequence 16S rRNA mitochondria DNA. The Bayesian phylogenetic tree was constructed using BEAST 2.4. and visualization in Figtree 1.4.4 (tree.bio.ed.ac.uk/software/figtree/). The best model of evolution was carried out using jModelTest 2.1.7. The most optimal was the evolutionary models GTR + I + G with p-inv (I) 0,1990 and gamma shape 0.1960. The main result indicated that the Bali cattle were grouped into Bos javanicus. Phylogenetic analysis also successfully classifying Bos javanicus, Bos indicus, Bos taurus, Bos frontalis and Bos grunniens. These results will complete information about Bali cattle and useful for the preservation and conservation strategies of Indonesian animal genetic resources.

Monomorphic allele rs109231213 in 3’UTR PLAG1 gene in purebred of Bali cattle (Bos javanicus)

The mutation rs109231213 that is located in 3’UTR of PLAG1 gene is associated with the growth and body weight in several Bos taurus and Bos indicus breeds. This study aimed to identify SNP rs109231213 in Bali cattle (Bos javanicus). The study used 41 samples of Bali cattle. The PLAG1 gene polymorphism was analyzed using PCR and direct sequencing methods. PCR pimers were 5’- TTGCACAGAATCAGTGTGTC-3’ and 5’- AGCCTAACGTGGATCTATGG-3’. The results showed that primers successfully amplified the 331 bp fragment at annealing 60°C that contained rs109231213. SNP was monomorphic in Bali cattle with one allele (G). This study concludes that rs109231213 in 3’UTR of PLAG1 gene can be used as specific marker in purebred of Bali cattle that have never been crossed with Bos taurus and Bos indicus.

Meta data analysis of conception rate in relation to sperm motility in Madura superior bulls

Madura bulls are Indonesian germplasm with a very high capacity to adapt to dry environments. Madura bulls come from a crossbreed between Zebu (Bos indicus) and banteng (Bos javanicus). One of the breeding strategies of Madura cattle is the use of artificial insemination (AI) with frozen semen. Regarding sperm motility as one of the standard parameters of good semen quality, it is good to know the reliability of sperm motility with the bull fertility rate. This study aimed to determine the conception rate percentage (%CR) relation to sperm motility in Superior Madura bulls. The frozen semen from eight Madura bulls belonging to the National Singosari and Lembang AI centre were used. They were classified based on the selected field reproductive efficiency data from the year 2018 until 2020. Sperm motility was evaluated using Computer Assisted Sperm Analysis (CASA). The data were analyzed using oneway ANOVA and Pearson correlation. The data showed that %CR was significantly higher (P<0.05) and positively correlated with sperm motility. It is proved that sperm motility represents good quality sperm as one of the fertility parameters in Madura bulls.

Fluctuating Asymmetry Increases with Heat Stress Disruptions on Bali Cattle (Bos javanicus) at Different Altitude

<p>Bali cattle (<em>Bos javanicus</em>) are common species distributed throughout Indonesia to survive in tropical hot, and humid climates. Fluctuating asymmetry (FA) is a random deviation from perfect bilateral symmetry. FA is often used to measure developmental stability in individuals and can be used as a marker for the impact of environmental and genetic stress. This study aims to investigate the pattern of developmental instability in Bali cattle as caused by environmental stress at various altitudes using the FA index. FA indexes in this study were used to compare asymmetrical physical traits of Bali cattle in Sembalun high altitude and Serading low altitude. Sixty-five Bali cattle were used in this study reared at different altitudes, namely Serading, Sumbawa Island (50 m above sea level), and Sembalun, Lombok Island (1,186 m above sea level). The physical traits of Bali cattle measured were horn circumference (horn), a distance of hip to spine bone (pelvic), metatarsal circumference (metatarsal), and metacarpal circumference (metacarpal). The FA1 and FA5 indexes showed significant metacarpal differences between Bali cattle reared at Sembalun and Serading population (P&lt;0.05). FA11 multiple trait index as a composite of all traits revealed a significant difference as well. (P&lt;001). It can be concluded that various heat stress by altitude impacts the asymmetry of Bali cattle.</p>

Hunting pressure is a key contributor to the impending extinction of Bornean wild cattle

Widespread and unregulated hunting of ungulates in Southeast Asia is resulting in population declines and localised extinctions. Increased access to previously remote tropical forest following logging and changes in land-use facilitates hunting of elusive wild cattle in Borneo, which preferentially select secluded habitat. We collated the first population parameters for the Endangered Bornean banteng Bos javanicus lowi and developed population models to simulate the effect of different hunting offtake rates on survival and the recovery of the population using reintroduced captive-bred individuals. Our findings suggest that the banteng population in Sabah is geographically divided into 4 management units based on connectivity: the Northeast, Sipitang (West), Central and Southeast, which all require active management to prevent further population decline and local extinction. With only 1% offtake, population growth ceased in the Northeast and Sipitang. In the Southeast and Central units, growth ceased at 2 and 4% offtake, respectively. Extinction was estimated at 21-39 yr when offtake was 5%, occurring first in Sipitang and last in the Central unit. Supplementing the population with captive-bred individuals suggested that inbreeding was likely to limit population growth if using ≤20 founder individuals. Translocating 2 individuals for a 10 yr period, starting after 20 yr of captive breeding resulted in a faster population recovery over 100 yr and a lower extinction probability. Our results suggest that shielding the population against further losses from hunting will be key to their survival in the wild, provided that active management in the form of captive breeding is developed in the interim.

Identification of Somatic Antigens of Adult Fasciola gigantica Isolated from Bali Cattle

In most tropical countries, such as Indonesia, fasciolosis is generally caused by Fasciola gigantica known as tropical liver fluke. However, most fasciolosis serodiagnostic tests have been developed solely for diagnosing fasciolosis caused by Fasciola hepatica (non-tropical liver fluke), and very few have been specifically designed for F. gigantica. The aim of this study was to determine the profile of antigenic proteins from the somatic extract of F. gigantica isolated from Bali cattle (Bos javanicus). The liver flukes were collected from a slaughtering house in Mataram, Indonesia. The somatic extracts were prepared by homogenizing in buffers containing 0.05 M NaCl, 0.02 M PMSF, and 0.05% Triton X-100. The characterization of the somatic extract proteins was performed using one-dimension gel electrophoresis and followed by Western blotting to determine the profile of its antigenic proteins. There were 14 bands of the somatic extracts with an estimated molecular weight ranging from 8 to105 8 kDa shown on the gel electrophoresis. The results of the Western blot show that there were five prominent protein bands. Three out of five prominent antigenic proteins with molecular weights of 8, 27, and 33 kDa are promising to enrich the existence of antigens that have immunodiagnostic value for fasciolosis. Therefore, further studies are required to examine more deeply the potency of those three antigenic somatic proteins of F. gigantica.

Phylogenetic Study of Madura Cattle Based on Mitochondrial Cyt b and D-loop Sequences

Madura Cattle is one breed of local cattle from Indonesia. Madura cattle are estimated to originate from a crossbreeding between Bos indicus and Bos javanicus. Another presumption is that Madura cattle are the result of a crossbreeding between B. indicus males and mixed B. javanicus or Bos taurus. Tracing the history of Madura cross and another cattle phylogenetic based on maternal lineage can be done by analyzing the variation of the mitochondrial genome (mtDNA). The purpose of this study was to determine the clarity of the origin of Madura cattle based on maternal lineage using mtDNA markers Cyt b and D-loop. This research is expected to provide genetic information and the origin of Madura cattle, so that it can be used to help improve the breeding and conservation program for Madura cattle. The results of the phylogeny tree reconstruction, using the Cyt b and D-loop genes showed that Madura cattle originated from Sampang region (Polagan, Golbung, and Komis) were grouped into two types of maternal origin. Madura cattle clade I are grouped with B. indicus and B. taurus, while Madura cattle clade II are grouped with B. javanicus. A crossbreeding between B. javanicus and B. indicus is estimated to have been carried out since the entry of Hindu culture brought by the India peoples to Indonesia around 1800 years ago. The crossing between B. javanicus and B. indicus was then more intensively carried out at the time of the government's promoting the development of Ongol cattles (B. indicus) in the days of the Dutch East Indies. The length segment of Cyt b that can be amplified is 230 bp and the D-loop segment of varying length, 577 bp for the Madura 41 and 29 samples, and 624 bp for sample 32.

Recolonization of Former Range by Endangered Banteng Bos javanicus in Mae Wong National Park, Thailand

Many large ungulate species, including the banteng ( Bos javanicus), are on the verge of extinction. Mae Wong National Park in Thailand was once a haven for banteng due to its abundance of preferred habitat, but the species was extirpated in the 1970s due to logging and hunting, before park designation. Mae Wong is connected with Huai Kha Khaeng Wildlife Sanctuary (HKK), which retains the largest banteng population remaining in Thailand. In 2019, Mae Wong park rangers reported suspected banteng footprints near HKK. We used camera traps to verify the presence of banteng and determine their population structure and activity pattern. We observed at least nine individual bantengs (range 1–7 each month) during the 15-month survey, including three adult males, two adult females, three juveniles, and 1 calf. Solitary males and herds (up to five animals) were observed, reflecting typical social structure. Banteng were detected throughout the survey and across all seasons, suggesting a resident population. Our research confirms the return of banteng to Mae Wong after an absence of over 40 years. The banteng likely dispersed from HKK, emphasizing the importance of landscape connectivity and source sites for recovery of endangered species. Though still small, the Mae Wong population is breeding, which is significant for future population recovery. Protection, community outreach, and habitat management can help secure this population. As banteng is the preferred food of tigers, their recovery will benefit tiger conservation in Mae Wong as well.

Entrapment Formulation for env-Tm Gene Based on Chitosan Low Molecular Weight as a Jembrana Disease Virus Vaccine Candidate

Jembrana disease (JD) caused by Jembrana Disease Virus (JDV) becomes an obstacle in Bali cattle (Bos javanicus). The development of JD vaccines has a critical meaning to prevent losses in the B. javanicus in Indonesia and is one of the models in the development of the Human Immunodeficiency Virus (HIV) vaccine. The development of vaccines for JDV has carried out DNA vaccines that are expected to provide better immunological effects. This study aimed to determine the low molecular weight chitosan (LC) entrapment towards pEGFP-C1-env-Tm in the formation of Chitosan Nanoparticles Low/pEGFP-C1-env-Tm complex. The env-Tm gene was inserted in pEGFP-C1 into the pEGFPC1/ env-Tm construct transformed on the E. coli DH5α host. The construct was formulated into LC/pEGFP-C1/env-Tm complex with a low molecular weight chitosan concentration of 0.06 % and the ratio of pEGFP-C1/env-Tm: LC (wt/wt) was 1:0.5-1:3. The complexes were then analyzed by gel retardation assay agarose 1 %. The results of this study indicated that the best entrapment results of low molecular weight chitosan to pEGFPC1/ env-Tm was in the mass ratio of pEGFP-C1/env-Tm: LC was 1:2. The best formulation entrapment for env-Tm by low molecular weight chitosan 0.06 % is in the rate 1:2.