CRISPR-associated proteins and clustered regularly interspaced short palindromic repeats (CRISPR-Cas) technology has emerged as a groundbreaking advancement in animal and poultry nutrition to improve feed conversion efficiency, enhance disease resistance, and improve the nutritional quality of animal products. Despite significant advancements, there is a research gap in the systematic understanding and comprehensive use of the CRISPR-Cas method in animal and poultry nutrition. The purpose of this study is to elucidate the latest advancements in animal and poultry nutrition through CRISPR-Cas genome editing technology, focusing on gene manipulation in metabolism, immunity, and growth. Following preferred reporting items in meta-analysis and systematic reviews guidelines, we conducted a systematic search using several databases, including Scopus, PubMed, and Web of Science, until May 2024, and finally, we included a total of 108 articles in this study. This article explores the use of the CRISPR-Cas system in the advancement of feed additives like probiotics and enzymes, which could reduce the use of antibiotics in animal production. Furthermore, the article discusses ethical and regulatory issues related to gene editing in animal and poultry nutrition, including concerns about animal welfare, food safety, and environmental impacts. Overall, the CRISPR-Cas system holds substantial promise to overcome the challenges in modern animal agriculture. By enriching the nutritional quality of animal products, increasing disease resistance, and improving feed efficiency, it offers sustainable and cost-effective solutions that can revolutionize animal and poultry nutrition [J Adv Vet Anim Res 2024; 11(2.000): 483-493]

Objective: Meatballs are a popular meat-based food consumed widely in Indonesian society. However, the issue of unethical substitution of halal meatballs with non-halal meats, particularly pork and canine meat (CM), has emerged. The existence of non-halal meats, including CM, in food products is prohibited in Islam, necessitating the development of reliable analytical techniques for their identification. In this study, we designed species-specific primers (SSPs) targeting the D-loop region of mitochondrial DNA for CM meatball product identification. Materials and Methods: The study was commenced by creating specific primers for canine DNA using Integrated DNA Technologies software and subsequently performing DNA isolation. The designed primers were then subjected to comprehensive evaluation using RT-PCR, including spec¬ification, linearity, limit of detection, efficiency, and repeatability. Results: The results indicated that the primer D-Loop 443 (forward: 5¢-GGG ACA TCT CGA TGG ACTA ATG-3', reverse: 5'-GCG GTC ATA GAT GAG TGA TAG C-3') designed and validated in silico using primer-basic local alignment search tool nucleotide (BLAST) program from NCBI accurately identified canine DNA when the optimal annealing temperature was set at 57.5oC. The real-time PCR technique utilizing the D-loop 443 primer exhibited the ability to amplify canine DNA down to a minimum quantity of 100 pg, with an efficiency value of 91.8%, a correlation coefficient (R) of 0.990, and a precision value (RSD) of 0.30%. Conclusion: The SSP-based RT-PCR method developed is a versatile and efficient tool for detect¬ing CM in meatballs. Its implementation helps maintain consumer trust and addresses concerns regarding the substitution of halal meats with non-halal alternatives. [J Adv Vet Anim Res 2024; 11(2.000): 247-258]

Objective: The aim of this investigation was to examine the impact of Taraxacum coreanum (known as dandelion) (TC) and TC mixtures with milk thistle (MT) or Aspergillus oryzae (AO) as feed additives on the immune response, milk quality, and milk production in Holstein cows over 6 weeks of administration. Materials and Methods: Thirty-two healthy Holstein dairy cows were provided 30 kg of total mixed ration (TMR) with no TC, 90 gm TC, 54 gm TC + 36 gm MT, or 54 gm TC + 36 gm AO 40% groups. The feed additives were supplied daily in two equal portions (per 45 gm) by topdressing the TMR for 6 weeks. Milk and blood samples were collected weekly. Results: In the TC-treated cows (TC, TC + MT, and TC + AO groups), significantly lower peripheral blood white blood cell (WBC) counts at 6 weeks and milk somatic cell counts (SCCs) at 4–6 weeks of administration were observed. Concentrations of serum superoxide dismutase (SOD) and glu¬tathione peroxidase (GSH-px) were notably elevated in cows treated with TC for 4–6 weeks, while levels of proinflammatory cytokines concentrations of tumor necrosis factor-alpha (TNF-α) and chemokine (IL-8) were significantly reduced in TC-treated cows after 3–6 weeks of administration. Conclusion: These results suggested that TC or a TC mixture with other medicinal herbs supple¬mentations enhanced the serum antioxidative activities and, consequently, might suppress the adverse immune response due to lower serum TNF-α and IL-8 release supported by lower WBC and SCC counts. [J Adv Vet Anim Res 2024; 11(2.000): 330-338]

Objective: The goal of this research was to evaluate where selenium nanoparticles impact the activity of antibodies in immunized lambs with foot and mouth vaccines by modulating the immune system. Materials and Methods: Two groups of lambs of 3–4 months of age were injected with 1 ml of ARRIAH-VAC vaccine intramuscularly in the neck, five Lambs were given selenium nanoparticles (size 100 nm) oral administration of selenium nano dose of 0.1 mg/kg of body mass once every day for sixty days considered as group one (G1) while the other five used as control Group 2 (G2). Results: This resulted in the establishment of an immune response, as evidenced by a rise in antibody titer in the blood using the ELISA test for three serotypes A, O, and Asia 1, when sele¬nium nanoparticles were given orally at a dose of 0.1 mg/kg body weight after immunization, we noticed a significant (p >0:05) selenium nano group increase in IgG response in all immunized groups in contrast to lambs that had only received the foot-and-mouth disease vaccine Conclusion: We have demonstrated that selenium nanoparticles administered orally significantly enhance immune responses while also increasing body weight. [J Adv Vet Anim Res 2024; 11(2.000): 367-375]

Objective: This study aimed to evaluate rumen fermentation parameters influenced by both the graz¬ing system and breed. Materials and Methods: A 2 × 2 factorial design was employed, involving 40 cows with matched age, parity, and physiological status. The cows were evenly divided between Bonsmara and Nguni breeds, as well as communal and commercial grazing systems. Rumen fluid samples were collected and analyzed for parameters including ammonia-nitrogen (NH3-N), pH, temperature, and volatile fatty acids (VFAs). Results: Nguni cows exhibited significantly higher ruminal NH3-N levels (p < 0.05) compared to Bonsmara, ranging from 69.05 to 96.78 mg/l. Commercial grazing demonstrated significantly higher NH3-N concentrations (p < 0.05) than communal grazing. Ruminal pH, temperature, total VFAs, and specific VFAs (Iso-butyrate, valeric, and iso-valeric) did not show significant differences (p > 0.05). However, total VFAs were slightly lower in communal grazing (78.87 mmol/l) than in commercial grazing (89.80 mmol/l). Acetate, propionate, butyrate, and the acetate to propionate ratio did not display significant differences (p > 0.05) between breeds but varied between grazing systems. Communal systems had higher acetate and acetate to propionate ratio (p < 0.05), while commercial systems showed higher propionate and butyrate levels (p < 0.05). Conclusion: Grazing conditions significantly influenced rumen fermentation parameters, irrespec¬tive of breed. Further research is necessary to explore the relationship between forage conditions, diversity, and rumen fermentation within different grazing systems. [J Adv Vet Anim Res 2024; 11(2.000): 349-355]

Objective: This study aimed to analyze the compound content of the combined extracts of Langusei fruit (Ficus minahassae L.) and papaya latex (Carica papaya L.) and perform bioassays of the combination of extracts on fleas of the genus Ctenocephalides. Materials and Methods: Liquid chromatography-tandem mass spectrometry (LC-MS/MS) determined the chemical composition of the combined extract. The toxicity of the extract combination was evaluated in vivo on cat flea and dog flea imago separately. The combination of phytochemical screening of Langusei extract and papaya latex contained all the main phytochemical compounds. Results: The LCMS/MS analysis results showed that the combination of Langusei extract and papaya latex contained six compounds. Four compounds were identified: 3-butenyl glucosinolate, erythromycin, aluminum palmitate, and henpentakontilbenzene. Four compounds indicate a new compound. The toxicity of the combined extract was highest for both Ctenocephalides felis and Ctenocephalides canis in the P4 treatment (10%), with an average mortality of 100%, the same as the control mortality of the synthetic insecticide deltamethrin. The best LD50 for C. felis was in the F1 formula (4.003 mg/l), while in C. canis it was shown in the F3 (3.733 mg/l). Conclusion: Based on the results obtained, the combination of Langusei fruit extract and papaya latex can be developed as an ectoparasiticide for fleas of the genus Ctenocephalides. [J Adv Vet Anim Res 2024; 11(3.000): 810-818]

Objectives: The study aimed to detect Neospora caninum by nested PCR (nPCR) in aborted fetuses of cattle, sheep, and goats in Bangladesh. Materials and Methods: The head portion of each aborted fetus (111) was dissected at each sampling site and transferred to the laboratory in an ice box. Data on risk factors associated with N. caninum infection were simultaneously collected. Deoxyribonucleic acid was extracted from brain tissue to perform nPCR targeting the internal transcribed spacer 1 (ITS1) ribosomal DNA (rDNA) gene of N. caninum and sequencing was performed from the representative positive samples. Results: By nPCR, N. caninum was found in 16.0% of aborted fetuses of cattle, followed by sheep (14.81%) and goats (11.78%). The highest prevalence was found in aborted fetuses of animals during the second trimester (27.78%) of pregnancy aged 2 to 4 years (18.75%). Obtained sequences showed they were completely matched with N. caninum ITS1 rDNA gene deposited in GenBank. Univariate analysis demonstrated that pregnancy stages (trimesters), abortion history of the animals, and access to dogs in animal farms were significantly (p ≤ 0.05) correlated with N. caninum infection. Conclusion: This study represents the first investigation into the molecular detection, phylogenetic characterization, and analysis of risk factors associated with N. caninum in livestock in Bangladesh. According to the research findings, N. caninum infection may have a role in abortion cases and the ensuing financial losses in the nation's livestock industry. [J Adv Vet Anim Res 2024; 11(3.000): 618-626]

Objectives: This study aims to determine the effect of a toll-like receptor 7/8 (TLR7/8) ligand on the motility of putative X- and Y-chromosome-bearing sperm in rams. Materials and Methods: Sperm from three fertile rams were incubated with tris-citrate buffer containing 0 to 0.9 μM resiquimod (a TLR7/8 ligand) that affects only the X chromosome sperm. Sperm was then subjected to a swim-up test method. After incubation, sperm in the top and lower layers of the media were transferred into a separate new tube. Sperm motility characteristics, concentration, morphometry, and adenosine triphosphate (ATP) concentration from both layers were identified. The data were analyzed for multiple comparisons using one-way analysis of variance. Results: The study demonstrated that the presence of resiquimod used in the swim-up method facilitated the morphometric separation of smaller sperm, predominantly representing Y chromosome sperm in the top layer. Meanwhile, larger samples, indicating X chromosome sperm, accumulated in the lower layer of the media, thus increasing sperm concentration. Resiquimod also decreased motility in the lower layer, but samples in the top layer were unaffected. This decrease was reinforced by the depletion of the ATP level of sperm at the lower level. Conclusion: The TLR7/8 ligand reduced the motility of the putative X-sperm by decreasing their ATP content, allowing separation from the putative Y-sperm. These results suggested the importance of TLR7/8 as a potential biomarker in sperm selection technology. [J Adv Vet Anim Res 2024; 11(3.000): 648-654]

Research articles about the effects of rumen-protected choline (RPC) supplementation on metabolic response, and reproductive and productive performances in transitional dairy cows were reviewed and presented. Analysis was conducted on 32 research papers that were published. The papers examined treatments without RPC supplementation and RPC supplementation levels varying from 10 to 100 gm/day. The feeding duration of RPC started from 40 to 140 days prepartum and continued until 20 to 140 days postpartum in multiparous dairy cows. Studies indicated that adding herbal choline (Cho) to the diet of transition dairy cows resulted in increased milk production and improved milk quality, leading to enhanced energy balance and reduced oxidative stress. The concentration and yield of IgG in colostrum provide passive immunity to dairy newborns and can be enhanced by dietary Cho supplementation. The inconsistent effects of RPC supplementation on reproduction may be due to several factors such as heat stress, genetics, and management. RPC supplementation improved the transition dairy cows' milk yield and quality, but dosage response was not observed as in the prior publication. Remarkably, the length of RPC supplementation had a positive correlation with an increase in milk yield. Based on this review, 45–50 gm/day dietary RPC supplementation between 3 weeks pre-calving to 8 weeks post-calving is suggested to increase at least 10% milk yield in dairy cows. [J Adv Vet Anim Res 2024; 11(3.000): 754-761]

Objective: The methodology employed in this research was designed to identify and characterize the infectious bursal disease virus (IBDV) at the molecular level, originating from recent outbreaks in Bangladesh. Materials and Methods: The IBDV outbreak farm was investigated, and bursa of Fabricius (BF) specimens were acquired from infected chickens. Initially, viruses in the processed samples were detected in chicken embryo fibroblast (CEF) cells, and the RT-PCR method was used to confirm IBDV. The positive samples were injected through chorioallantoic membrane route into the embryo of a 10-day-old specific pathogen-free (SPF) egg for virus isolation and pathogenicity testing. Finally, we sequenced the VP2 gene to identify phylogenetic relationships and detect mutations. Results: From the 77 collected samples, 42.85% (33/77) were found positive for cytopathic effects in CEF cells, and IBDV was detected in 31.16% (24/77) of the samples by RT-PCR. IBDV was isolated in SPF chicken embryos. In the pathogenicity test, infectious bursal disease was evident in seronegative chickens with visible signs of disease. Sequence analysis shows that the broiler-isolated viruses clustered with genotype A3B2 and backyard chickens with genotype A1B1. The presence of amino acid motifs for virulence markers was revealed in the partially sequenced VP2 gene with a mutation at S254G in four IBDV isolates from broilers. However, amino acids for virulence markers were absent in two isolates from backyard chickens, which shows sequence homology with IBDV classic strains. Conclusion: In this study, we identified and characterized circulating reassorted IBDV from vaccinated broilers, which may be one of the major causes of vaccination failure in broilers. [J Adv Vet Anim Res 2024; 11(3.000): 534-543]