Bajaree Chuttong¹, Supakhom Klaitanoad¹, Kanokwan Klaithin¹, Sampat Ghosh^2,3, Chuleui Jung^2,4, Ming Cheng Wu⁵, Khanchai Danmek^6*

¹Meliponini and Apini Research Laboratory, Department of Entomology and Plant Pathology, Faculty of Agriculture, Chiang Mai University 50200, Thailand

²Agricultural Research Institute, Gyeongkuk National University, Andong GB 36729, South Korea

³Department of Life Science Sardar Patel University, Balaghat, Madhya Pradesh 481331, India

⁴Department of Plant Medicals, Gyeongkuk National University, Andong GB 36729, Republic of Korea

⁵Department of Entomology, College of Agriculture and Natural Resources, National Chung Hsing University 402202, Taiwan

⁶Biotechnology program, School of Agriculture and Natural Resources, University of Phayao 56000, Thailand

*Corresponding author’s email: khandchai.da@up.ac.th

Received: 25 January 2025 / Accepted: 23 May 2025 / Published Online: 31 May 2025

Abstract

This study investigates the impact of Aspergillus flavus, a mycotoxin-producing fungus, on honeybee health in Thailand, where maize pollen is a key nutritional resource for apiculture but carries a significant risk of fungal contamination. Metabolomic analysis revealed that A. flavus produces aflatoxins B1 (AFB₁) at 3,753 ng/mL and B2 (AFB₂) at 327 ng/mL, while A. nomius is not toxin-producing. Co-culturing A. flavus with A. nomius led to a reduction in AFB₁ and AFB₂ to 218 and 26.52 mg/mL, respectively. Newly emerged worker honeybees were fed various diets: 50% sucrose syrup (S, T1), S + semi-synthetic media (T2), S + metabolites of A. flavus (T3), S + metabolites of A. nomius (T4), and S + metabolites of both A. flavus and A. nomius (T5). The results indicated that T3 resulted in the shortest lifespan and the smallest hypopharyngeal gland acini size (9.38 ± 1.02 nm compared to 21.07 ± 1.37 nm in T2). However, the lifespan of T4 and T5 was comparable to T1 and T2, indicating that A. nomius would not have any harmful impact on honeybee lifespan and may reduce any deleterious impacts of A. flavus. Similar inference was possible on the acini size of the hypopharyngeal gland which is an important health parameter of worker honeybees. This study contributes to a deeper understanding of how microbial contamination affects honeybee health and the necessity to reduce the risks associated with toxin exposure under the situation of natural pollen sources declining.

Keywords: Fungal contamination, Honeybee health, Hypopharyngeal glands, Maize pollen, Toxin exposure

Aparicio-Ortiz Mónica¹, Juárez-Monroy Dilery¹, Canto-Canché Blondy², Tzec-Simá Miguel¹, Islas-Flores Ignacio^1*

¹Unidad de Biología Integrativa, Centro de Investigación Científica de Yucatán, A.C., calle 43 No. 130 x 32 y 34, Chuburná de Hidalgo, C.P. 97205, Mérida, Yucatán, México

²Unidad de Biotecnología, Centro de Investigación Científica de Yucatán, A.C., calle 43 No. 130 x 32 y 34, Chuburná de Hidalgo, C.P. 97205, Mérida, Yucatán, México

*Corresponding author’s email: islasign@cicy.mx

Received: 28 November 2024 / Accepted: 23 May 2025 / Published Online: 31 May 2025

Abstract

Highly methyl-esterified pectin (HME-P) is typically found in the primary cell walls of young and embryonic plant cells, where it plays a role in regulating differentiation and maturation processes. In this study, the JIM7 monoclonal antibody, which specifically targets HME-P, was utilized to examine tissue sections of coconut zygotic embryos and solid endosperms from the Yucatan green dwarf cultivar at various developmental stages. The expression of the pectin methyl-esterase gene was also assessed using RT-qPCR. The JIM7 antibody revealed the presence of HME-P surrounding the inner cells of immature embryos at the globular and coleoptile stages. As the embryos progressed to intermediate and mature stages, HME-P was found in various cell types, including those of the plumule. In the immature solid endosperm, HME-P was associated with syncytial cells. At the intermediate stage, it was observed in vesicle-like structures and by the mature stage, it surrounded compact cells in a distinct mosaic pattern, near the corrosion cavity. Analysis of the coconut PME gene expression in solid endosperm and zygotic embryos showed peak expression at the immature stage, followed by a decline at the intermediate and mature stages. These findings were consistent with the immunodetection results obtained with the JIM7 antibody in embryos and endosperms. Overall, this study provides novel insights into the coordinated expression of the PME gene and the dynamics of HME-P during the maturation of coconut zygotic embryos and solid endosperms. These findings suggest a potential role for pectin methyl esterification in zygotic embryogenesis in coconut.

Keywords: Endosperm, Coconut, Embryo, Pectin, JIM7, Methyl-esterification

Yuexia Wang^1,2, Shiming Han^1,2*, Muhammad Shahr Yar Ali³, Shuai Zheng², Jihong Dong¹, Muhammad Sajjad^3*, Yumei Fang²

¹School of Public Administration, China University of Mining and Technology, Xuzhou, Jiangsu, China

²School of Biological Sciences and Technology, Liupanshui Normal University, Liupanshui, P.R. China

³Department of Biosciences, COMSATS University Islamabad, Islamabad 45550, Pakistan

*Corresponding authors’ emails: hanshiliang888@163.com; msajjadpbg@gmail.com

Received: 04 March 2025 / Accepted:20 April 2025 / Published Online: 28 May 2025

Abstract

Actinidia valvata Dunn is a fruit tree that originated in China and is well recognized for its nutrient, health, and medicinal value. With the aim of identifying the nutrients involved, this study conducted a comprehensive transcriptome analysis of the stem, leaf, and fruit of A. valvata using the Illumina HiSeq platform. RNA-Seq analysis produced 39.03 GB of clean data and further identified a total of 50,928 unigenes with an average length of 1,236.40 bp. Functional annotation of these unigenes was carried out by comparing them with several databases. This resulted in the incorporation of annotated information regarding 32,202 unigenes, which constituted 63.23% of the total. Moreover, 11,809 unigenes participated in 129 unique metabolic pathways. Comparative analysis of gene expression across various tissues led to the identification of differentially expressed genes (DEGs). Among these, 26 DEGs were closely associated with sugar metabolism pathways. Moreover, correlation analysis between total sugar content and expression of the DEGs implied in sugar metabolism proved that the identified key genes SUS, INV, SPS, HK, malZ, and GPI have strong functions in sugar metabolism in A. valvata. Our findings reveal that sucrose synthase (SUS) and invertase (INV) are critical drivers of sugar accumulation in fruits, providing targets for breeding sweeter kiwifruit varieties.

Keywords: Kiwi berry, A. valvata, Total phenolic, SUS, INV, SPS, DEGs, Sugar metabolism

Ahmed Raza^1,2,3*, Muhammad Taimoor Shakeel⁴, Sidra Iqbal⁵, Azhar Abbas Khan⁶, Tahira Abbas⁷, Ammarah Hasnain⁸, Mahmoud Ahmed Amer⁹, Abdul Majid², Adnan Ahmad¹⁰, Ummad Ud Din Umar¹¹, Muhammad Umer Iqbal¹², Muhammad Naveed Aslam⁴, Xifeng Wang³, Mingzheng Duan^1*

¹Key Laboratory of Edible Fungi Resources Innovation Utilization and Cultivation, College of Agronomy and Life Sciences, Zhaotong,657000, China

²Plant Virology Lab, Crop Diseases Research Institute, National Agriculture Research Centre, Pakistan Agricultural Research Council, Islamabad, Pakistan

³State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, China

⁴Department of Plant Pathology, Faculty of Agriculture & Environment, The Islamia University of Bahawalpur, Bahawalpur 63100, Pakistan

⁵Department of Plant Breeding and Genetics, University of Agriculture-Faisalabad, Constituent College Depalpur, Okara, 56300, Pakistan

⁶Department of Entomology, Faculty of Agricultural Sciences and Technology, University of Layyah, Pakistan

⁷Department of Horticulture, Faculty of Agricultural Sciences and Technology, University of Layyah, Pakistan

⁸Department of Biotechnology, Faculty of Biological Sciences, Lahore University of Biological and Applied Sciences, Pakistan

⁹Plant Protection Department, College of Food & Agriculture Sciences, King Saud University, P.O. Box 2460, Riyadh 11451, Saudi Arabia

¹⁰GenEd, Pakistan Council of Scientific and Industrial Research, Lahore, 54600, Pakistan

¹¹Department of Plant Pathology, Faculty of Agriculture Science and Technology, Bahauddin Zakariya University, Multan 60800, Pakistan

¹²Department of Plant Pathology, Muhammad Nawaz Sharif University of Agriculture, Multan, Pakistan

*Corresponding author’s email: ahmedraza7128@parc.gov.pk; duanmingzheng@ztu.edu.cn

Received: 13June 2024 / Accepted: 20 April 2025 / Published Online: 28 May 2025

Abstract

Tomato chlorosis virus (ToCV), causing yellowing disease in tomato, is a phloem-limited, whitefly-transmitted crinivirus that mainly affects plants by reducing chlorophyll content. During a survey conducted in 2019, cirinivirus-specific yellowing symptoms were observed in the tomato fields in Multan, Khanewal, and Lodhran districts of Pakistan. To confirm the etiology of the yellowing disease of tomato, a total of 76 symptomatic and 22 asymptomatic samples were collected out of which 72% of the tested plants were found to be positive for ToCV using RT-PCR. The full genomic nucleotide sequences of two selected ToCV-isolates were obtained by next-generation sequencing (NGS). RNA1 and RNA2 of each isolate comprised 8594 and 8242 nucleotides, respectively. The NGS results were verified by sequencing the amplified overlapping fragments of RNA1 and RNA2 using specific primers designed in this study. In BLASTn analysis for ToCV-Pak1 and ToCV-Pak2, RNA1 from both isolates had the highest similarities (99.41-99.46%) with a South Korean isolate; RNA2 had the highest similarities of 99.67-99.64% with a Greek isolate for Pak1 and Turkish isolate for Pak2, respectively. Phylogenetic analysis indicated that RNA1 of the Pakistani isolates clustered together, forming a subclade, and RNA2 from one of the Pakistani isolates (MN869006) clustered with a Brazilian isolate. At the same time, the other one (MN869007) fell close to a Turkish isolate. Statistical analysis indicated a low polymorphic frequency in RNA1 but a higher polymorphic frequency in RNA 2 quantifying the degree of variation in the analyzed isolates which may result in emergence of new strains with altered levels of virulence. One probable recombination event was detected in RNA1. Analysis revealed that RNA1 of Pak1 and Pak2 is recombinant between South Korean isolate (major parent) and Spanish isolates (minor parent) with recombinant breakpoints at 6172 and 6668 nucleotide positions from the 5′ end, respectively. This study provides the first full genomic analysis and the genetic diversity of ToCV-RNA1 infecting the tomato plant in Pakistan. Understanding the trends and rate of variation in the isolates may provide an insight into the development of resistance in tomato plants against tomato yellowing disease.

Keywords: ToCV, Phylogenetic analysis, Recombination, Genetic characterization, Tomato

L.S. Huseynova¹*, S.N. Mammadova², L.M. Suleymanova², G.A. Valiyeva²

¹Department of Natural Sciences, Western Caspian University, Baku, Azerbaijan

²Department of Medical Biology and Genetics, Azerbaijan Medical University, Baku, Azerbaijan

*Corresponding author’s email: royahuseynova2006@gmail.com

Received: 13 January 2025 / Accepted: 15 May 2025 / Published Online: 24 May 2025

Abstract

Mutations in BCKDHA, BCKDHB, DLD and DBT are associated with Maple Syrup Urine Disease (MSUD) in different populations worldwide, therefore screening of these genes is crucial for genetic counseling, diagnosis and treatments of MSUD in patients suffering from MSUD. The present study was therefore carried out by screening MSUD patients from different regions including Baku, Sheki-Zagatala, Guba-Khachmaz, Lankaran-Astara zones to establish the genetic spectrum of MSUD patients of in Azerbaijan population. The cohort consisted of 800 patients clinically diagnosed with MSUD from the year 2015 to 2020. The gene panel sequencing (BCKDHA, BCKDHB, DLD and DBT) was performed by amplification of exonic sequences using the polymerase chain reaction (PCR) method followed by Sanger sequencing of the amplified product. For the confirmation of identified mutation a control panel of 140 healthy individuals was also collected from the population. The sequencing of the exonic sequence of the four genes resulted in identification of four missense variants. One patient carried compound heterozygous variants 1221(A>G) and 972 (C>T) in exon 9 and 10 respectively while another patient had homozygous 508(C>T) variant in BCKDHB, and homozygous 1199(A>G) was found in DBT in another patient. All the three patients carrying the identified mutations were from Guba-Khachmaz zone: one was Azerbaijani Turk, and two were Lezgi ethnic group. The in silico analysis predicted the variants to be pathogenic for protein function. In conclusion, the present study highlights the genetic role of BCKDHB and DBT in onset of MSUD in Azerbaijan especially Guba-Khachmaz zone where BCKDHB variants found to be more common in MSUD patient with followed by DBT. The identified variants were absent from the control group, thus suggesting their role in disease manifestation. Out of the 800 MSUD screened patients excluded cases might be having deep intronic mutation either in BCKDHB and DBT or other gene or might be having novel gene which could only be identified through exome/genome sequencing of the remaining patient.

Keywords: MSUD, Genetic spectrum, Azerbaijani Turk, Guba-Khachmaz zone, Rare mutations, Ethnic specificity

Naeema A. Yahya^1,4*, Mohamed A. Awad², Shafik D. Ibrahim³, Mohamed A. Nagaty²

¹Genetic and Cytology Unit, Department of Genetic Resources, Desert Research Center, Cairo, 11753 Egypt

²Department of Plant Production, Faculty of Environmental Agricultural Sciences, Arish University, North Sinai, Egypt

³Agricultural Genetic Engineering Research Institute (AGERI), Agricultural Research Center, Giza, Egypt

⁴Plant Research Department, Agricultural Research Department, Ministry of Municipality, Doha, P.O. Box 2727, Qatar

*Corresponding author’s email: nabdelhalem@mm.gov.qa

Received: 28 December 2024 / Accepted: 06 May 2025 / Published Online: 24 May 2025

Abstract

This study used 38 morphological characteristics, IRAP and SCoT markers, and DNA barcoding to assess the relationships among seven date palm cultivars commonly grown in North Sinai, Egypt—five classified and two unknowns. Results of morphological characterization analysis within date palm cultivars showed significant variability (P ≤ 0.05) in tree, leaf, fruit, and seed morphological traits. Principal component analysis revealed strong relationships among certain morphological traits and cultivars, forming four major homogeneous variable groups. Cluster analyses revealed that Beid El Gamal exhibited the lowest genetic similarity among the seven cultivars, while the highest similarity index was observed between Amri and Yellow Majhal, Hayani and Red Majhal, and Khedri and Kapoushi. The evaluation was performed using IRAP and SCoT markers; IRAP markers generated 97 amplicons with 36% polymorphism, while SCoT markers produced 89 amplicons with 45% polymorphism. Genetic similarity coefficients ranged from 0.83 to 0.93, with the highest similarity (0.93) observed between Amri and Yellow Majhal cultivars. DNA barcoding using rbcL, matK, and trnH genes successfully differentiated the cultivars into distinct phylogenetic groups. Molecular analyses revealed that Amri and Yellow Majhal cultivars were most closely related, while Beid El Gamal showed the lowest similarity. We concluded that analyzing morphological traits can distinguish date palm cultivars, but it is time-consuming and requires specific traits to identify each cultivar. By integrating morphological and genetic characteristics, we achieved more reliable identification. Molecular markers, such as IRAP and SCoT—particularly using IRAP-2204 or SCoT-09 primers—along with DNA barcoding, allow for quick and accurate differentiation of closely related cultivars, even unknown ones. This approach enhances sustainable date palm production by conserving genetic resources, improving breeding programs, verifying identities, tracking genetic diversity, and protecting local cultivars.

Keywords: Date palm, Morphological identification, DNA barcoding, Genetic diversity, IRAP markers, SCoT markers

Sirinda Sutthachai^1,2, Attachai Trunjaruen³, Ploinapat Mahatthanaphatcharakun^1,2, Worasitikulya Taratima^1,2*

¹Department of Biology, Faculty of Science, Khon Kaen University, Khon Kaen 40002, Thailand

²Salt-tolerant Rice Research Group, Faculty of Science, Khon Kaen University, 40002, Thailand

³Mahidol University, Amnatcharoen Campus, Amnatcharoen 37000, Thailand

*Corresponding author’s email: worasitikulya@gmail.com

Received: 19 December 2024 / Accepted: 05 May 2025 / Published Online: 24 May 2025

Abstract

Upland rice cultivation in highland regions is often challenged by drought, leading to poor seedling growth and low productivity. This study aimed to investigate the physiological, biochemical, and anatomical responses of the Thai rice landrace Nhonkhab under drought and rewatering conditions. After germination, 28-day-old Nhonkhab rice seedlings were deprived of water for 7 days (35 days old) and then rewatered for 5 days (40 days old). Growth, physiological, biochemical and anatomical characteristics of the 35 and 40-day-old seedlings were evaluated. Results showed that drought stress significantly reduced relative water content (RWC) (-36.38%) but was rapidly restored after rewatering (+12.15%) when compared to the control. After drought stress treatment, malondialdehyde (MDA) content and electrolyte leakage (EL) percentage increased (+150% and +59.37%, respectively), indicating oxidative stress and membrane damage. Growth and developmental adaptations, such as elongated roots and curled leaves, facilitated survival during drought, with leaves unfurling post-rewatering. Following rewatering, rice plants exhibited robust recovery, supported by internal structural adaptations such as increasing stomatal density, thickening of the cuticle layer around the midrib and leaf blades, increasing bulliform cells number and size, and modifications in vascular bundles. Principal component analysis (PCA) and hierarchical cluster analysis (HCA) revealed distinct patterns in growth, physiological, and anatomical traits across control and treatment groups. These findings provide valuable insights for rice production planning in drought-prone highlands and for selecting or breeding drought-resistant cultivars with improved resilience and productivity.

Keywords: Electrolyte leakage, Environmental stress, Leaf anatomy, MDA, Upland rice, Water deficiency

Nurah M. Alzamel^*

Department of Biology, College of Science and Humanities, Shaqra University, Shaqra 11961, Saudi Arabia

*Corresponding author’s emails: nurahalzamel@gmail.com; nalzamel@su.edu.sa

Received: 06 October 2024 / Accepted: 06 May 2025 / Published Online: 24 May 2025

Abstract

Irrigation with saline water significantly reduces maize cultivation and productivity worldwide. Therefore, it is essential to investigate sustainable approaches to mitigate diverse effects of irrigation with saline water on various physiological and agronomical characteristics of maize, such as foliar application of selenium and proline. Two experiments, consequently, were carried out in the experimental farm of the College of Sciences and Humanities, Shaqra University, Al-Quwayiyah Governorate, Kingdom of Saudi Arabia during the two successive autumn seasons of 2022 and 2023. Three saltwater irrigation treatments were separately applied using a randomized complete block design with a split-plot arrangement with three replications. In each one of these, three rates of selenium were distributed in the main plots, whereas three rates of proline were applied in the sub-plots. Selenium and proline applications, individually or combined, significantly reduced the effects of saltwater irrigation stress on physiological and agronomic traits of maize. Obviously, optimal results were achieved with selenium at 10 mg L^-1 and proline at 10 mM, effectively mitigating the adverse effects of saltwater irrigation on the physiological and agronomic traits of maize. At a moderate salinity level of 2000 mg L^-1 NaCl, the application of 5, and 10 mM proline improved grain yield per plant by 7.76 and 5.54 %, as well as 17.62 & 7.99%; grain yield per hectare by 7.18 and 7.72 %, along with 19.11 & 8.33 %; and stover yield per hectare by 8.56 and 6.87 %, as well as 16.38 and 9.05 %, in both seasons, respectively under the application of 5, and 10 mg L^-1 selenium, respectively, as compared to severe salinity level of 3000 mg L^-1 NaCl. This in turn showed that exogenous selenium and proline application, individually or combined, were effective strategies in mitigating the harmful effects of salinity on physiological and agronomic traits of maize.

Keywords: Grain yield, Maize, Proline, Saline irrigation stress, Selenium

Asim Sultan¹, Muhammad Kashif Saleemi¹, Muhammad Saqib², Shafia Tehseen Gul^1*

¹Department of Pathology, Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan

²Department of Clinical Medicine and Surgery, Faculty of Veterinary Science, University of Agriculture, Faisalabad, Pakistan

*Corresponding author’s email: drshafia66@yahoo.com; dr.shafia.gul@uaf.edu.pk

Received: 27 January 2025 / Accepted: 06 April 2025 / Published Online: 04 May 2025

Abstract

In the present study, Clostridium perfringens (C. perfringens) was isolated from clinically suspected cases of necrotic enteritis. For this purpose, the intestinal samples collected, after enrichment in thioglycolate broth were inoculated on perfringens-specific media (tryptose sulphite cycloserine agar) in an anaerobic jar having a gas pack. After incubation of 24hrs at 37°C, characteristic black colonies on TSC agar were used for confirmation by gram staining. Molecular confirmation was carried out with species-specific 16 s rRNA primer and DNA was amplified further for different toxin genes i.e. cpa, cpb, etx, iap, cpe, and net B for toxinotyping of C. perfringens. These isolates were further used to induce necrotic enteritis in poultry birds and for this purpose experimental trials were carried out to compare the ameliorative effects of antimicrobial alternatives i.e. Tibetan tea extract, black tea extract. and they were compared with commercial available AGPs. Data was analyzed through analysis of variance (ANOVA) through SAS University Edition. Results of the experimental trial demonstrated that there is a significant improvement in physiological performance and immunity of the birds supplemented with tea extracts and exposed to the NE challenge. The current study finding conclude that the both Tibetan and Black tea extracts can be used separately or in combination for the control of necrotic enteritis in poultry. Tea phenolics have proved their potential as an alternative to antimicrobials and to address antimicrobial resistance in poultry.

Keywords: Necrotic enteritis, Herbal tea extract, Immunity, Physical parameters

SM Azizul Karim Hussaini¹, Farhan Ibne Siddique², Mokammel Hossain Tito⁸, Abdullah Al Mamun¹, Md Arifuzzaman³, Afzal Haq Asif⁴, Shahzad Khan⁵, Muhammad Shahzad Chohan⁵, Samar Sindi⁶, Talha Yusuf ⁷, Md. Siddiqur Rahman¹*

¹Department of Medicine, Faculty of Veterinary Science, Bangladesh Agricultural University, Bangladesh

²Armed Forces Medical College, Dhaka Cantonment, Dhaka, Bangladesh

³Department of Civil & Environmental Engineering, King Faisal University, Saudi Arabia

⁴Department of Pharmacy Practice, College of Clinical Pharmacy, King Faisal University, Saudi Arabia

⁵Department of Biomedical Sciences, College of Clinical Pharmacy, King Faisal University, Saudi Arabia

⁶Department of Biological Sciences, Faculty of Science, King Abdulaziz University, Saudi Arabia

⁷Department of Computer Sciences, FCIT, King Abdulaziz University, Saudi Arabia

⁸Department of Medicine, Gazipur Agricultural University, Bangladesh

*Corresponding author’s email: siddique.medicine@bau.edu.bd

Received: 23 January 2025 / Accepted: 15 April 2025 / Published Online: 04 May 2025

Abstract

Brucellosis is a highly contagious zoonotic disease caused by the bacterium Brucella spp. that distresses mutually animals and humans, especially in underdeveloped countries with poor control programs. In adult cattle, the disease affects mainly the reproductive organs, thus causing major losses in production and reproduction, such as abortion and reduced fertility. This study involves the collection of 460 blood samples from dairy farms, which were analysed for brucellosis infection using the Rose Bengal Test (RBT). Additionally, data on the animals’ history, including placenta (retained), repeat breeding, their age, abortion, and lastly calving, were also recorded. To address the problem of class imbalance between the positive and negative classes, a technique, known as Synthetic Minority Over-sampling Technique (SMOTE) was applied in the research work. A total of five algorithms were used in this paper among them multilayer perceptron (MLP) and weekadeeplearning4j showed the best results for the prediction of brucellosis having 93.59% and 93.94% accuracy, respectively. Besides, risk factors are ranked based on their importance as ordered as retained placenta > repeat breeding > calving > abortion > age, and three association rules are made to understand the correlation of the factors for occurring the disease. By applying this study, early diagnosis of the disease could be possible to mitigate the economic losses.

Keywords: Brucellosis, Zoonotic disease, Dairy cattle, Machine learning, Risk factors, Public health