Category Archives: b_original_articles

Muhmmad Kashif Shahzad Sarwar^1,2, Abdul Ghaffar³, Saghir Ahmad⁴, Shoaib Ur Rehman¹, Ummara Waheed^1*

¹Institute of Plant Breeding and Biotechnology, MNS University of Agriculture, Multan, Pakistan

²Cotton Research Station, Ayub Agricultural Research Institute, Faisalabad, Pakistan

³Departemnt of Agronomy, MNS University of Agriculture, Multan, Pakistan

⁴Cotton Research Institute, Multan, Pakistan

Abstract

Cotton diversity has long been studied using physiological and biochemical traits. This diversity has led to the development of various superior cotton cultivars over the year. At present and in face of climate change, development of high yielding and drought tolerant cotton varieties are necessary to fulfill the demand of ever-growing population of the world. In this study, Gossypium hirsutum L. germplasm (200) was evaluated under two irrigation regimes i.e., well-watered (W1) and limited water (W2) conditions. Various morphological and physiological traits were recorded under both irrigation regimes. A considerable reduction was recorded in W2 conditions in all the recorded traits except for glycine betaine, soluble sugars, and proline contents, highlighting the impact of drought on cotton germplasm. Cotton genotypes that maintained higher yield had positive correlation with biochemical traits. Out of 63 best performing genotype (superior parents based on the recorded data), FH-414, FH-415, FH-416, FH-326, FH-492, FH-Anmol, Gomal-105, Marvi, NIAB-878 and VH-327 were selected for hybridization to make crosses following Line x Tester fashion. F₁ hybrids (25 crosses) and 10 parents were again planted under W1 and W2 conditions. Out of 25 crosses, FH-326 × Marvi (CS5) and NIAB-878 × FH-414 (CS16) performed better under water deficit conditions. Quantitative real-time PCR was also performed using GhHH3 and GhIDD. CS5 and CS16 had higher expression of drought tolerance causing GhHH3 and GhIDD genes. The newly developed cotton crosses will pave the way for the development of high yielding drought tolerant cotton varieties in face of climate change.

Keywords: Biochemical attributes, Yield and yield components, GhHH3, GhIDD, Drought

Usama bin Naeem¹, Muhammad Adil Rasheed^1*, Muhammad Ashraf¹, Muhammad Yasir Zahoor²

¹Department of Pharmacology and Toxicology, Faculty of Biosciences, University of Veterinary and Animal Sciences Lahore, Pakistan

²Insititue of Biochemistry and Biotechnology, Faculty of Biosciences, University of Veterinary and Animal Sciences Lahore, Pakistan

Abstract

One of the most dominant diseases in the world, particularly among women, is breast cancer. Breast cancer has tumor suppressor genes called CHEK2 and TP53. When there is a mutation in CHEK2 and TP53 genes there are more chances of breast cancer. This study aimed to investigate the already prepared and characterized nanoparticles loaded with Chitosan for Cell death, Mitochondrial Membrane and cell cycle arrest estimated through Flow Cytometry and gene expression analysis of CHEK2 and TP53 genes by real-time PCR. The Livak method was used to evaluate the results. The mean (± S.D) comparison between the control and target genes were used to calculate gene expression. Results showed that Ivermectin and Tamoxifen NPs (B+C) represented 34.8% cell death that is better than other combinations with propidium iodide stain while with Acridine orange stain Tamoxifen+Ivermectin (A+B) combination showed the remarkable and maximum of the all cell cycle arrest with value of 69.7% cell arrest at G0/G1 phase, 7.11% of cell arrest at S Phase and 7.05% of G2/M Phase arrest. It was demonstrated that the expression levels of CHEK2 and TP53 genes were significantly increased (P<0.001) in Ivermectin+Tamoxifen NPs (B+C) compared with control groups. It is concluded that Tamoxifen nanoparticles with Ivermectin showed strong anti-proliferative activity against breast cancer cells. The expression levels of nanoparticles containing Tamoxifen were significantly increased compared to the other treatments and control groups (P<0.001). Gene expression change with change in dose concentrations.

Keywords: Breast cancer, Apoptosis, Cell cycle arrest, Pharmacogenomic, Gene expression

Muhammad Khalid Mansoor*¹, Kashif Iqbal¹, Ali Hassan², Muhammad Saqib², Ali Zohaib¹, Sabiqaa Masood³

¹Department of Microbiology, Faculty of Veterinary & Animal Science, The Islamia University of Bahawalpur, Pakistan

²Department of Clinical Medicine & Surgery, University of Agriculture, Faisalabad, Pakistan

³Department of Parasitology, Faculty of Veterinary & Animal Science, The Islamia University of Bahawalpur, Pakistan

*Corresponding author’s email: khalid.mansoor@iub.edu.pk

Received: 13 September 2023 / Accepted: 5 April 2024 / Published Online: 13 December 2024

Abstract

Contagious Ecthyma (CE), also known as scabby mouth disease, is caused by an epitheliotropic parapoxvirus that primarily affects the goat and sheep populations worldwide. This study focused on investigating 12 outbreaks of CE in sheep and goat herds across various regions of Punjab, Pakistan. A total of 35 samples were collected between March 2021 and May 2022, with 34 out of 35 samples testing positive for parapoxvirus through PCR. Subsequently, 24 complete sequences of the major envelope protein B2L gene were successfully obtained. The nucleotide and amino acid sequences of the ORF virus B2L gene were analyzed. The 1206bp amplicons, after Sanger sequencing revealed an open reading frame of 1137bp encoding 378 amino acids. The minimum and maximum nucleotide differences of 0 and 34, respectively, were observed, while the percentage similarity at the nucleotide level and amino acid level ranged from 97.98% to 100% and 97.62% to 100%, respectively, among the ORF strains in this research study. The results of the phylogenetic analysis revealed that all 24 ORF virus isolates from Pakistan belonged to Group-I ORF viruses. The comparative homology of Pakistani ORF virus strains with Indian, Chinese, and Turkish isolates was 99.03%, 98.59%, and 98.15%, respectively. This study contributes to understanding the circulation of Group I ORF viruses in Pakistan and their relationship with strains from neighbouring countries. Furthermore, these findings may offer insights into the genotype of the causative agent responsible for the contagious pustular dermatitis (CPD) outbreak in Punjab, Pakistan.

Keywords: B2L gene, Contagious, Contagious pustular dermatitis, ORF

Gulnaz Afzal1*, Muhammad Irfan Ullah2, Nadeem Ali3, Moeen Afzal1, Riaz Hussain4, Nabil A Alhakamy5, Nisreen Rajeh6, Sarmad Rehan7, Rehana Iqbal8, Muhammad Shahid Iqbal1, Ahrar Khan9*
1Department of Zoology, The Islamia University of Bahawalpur 63100, Pakistan
2Department of Pathobiology, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan
3Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah 21589, Saudi Arabia
4Department of Pathology, Faculty of Veterinary and Animal Sciences, The Islamia University of Bahawalpur 63100, Pakistan
5Pharmaceutics Department, Faculty of Pharmacy, King Abdulaziz University, Jeddah 21589, Saudi Arabia
6Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah 21589, Saudi Arabia
7Department of Anatomy, Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan
8Institute of Pure and Applied Biology, Zoology Division, Bahauddin Zakariya University, Multan, Pakistan
9Faculty of Veterinary Sciences, University of Agriculture, Faisalabad, Pakistan

Abstract
Nanoparticles are used extensively in various industries, such as agriculture, food packaging, medical diagnostics and electronics. However, their increasing usage raises concerns regarding potential health hazards and environmental risks. This study examined the impact of intra-peritoneal injections of magnesium oxide (MgO) nanoparticles on the brain, testis, and muscles of male albino rats. Mature male rats (n=20) after acclimatization were randomly divided into four groups (G0, G1, G2, G3). The rats in the treated groups (G1-G3) were given MgO NPs @ 25 mg/kg, 50 mg/kg and 75 mg/kg respectively for ten consecutive days. G0 rats served as untreated control group. Results indicated that MgO NPs induced clinical alterations in exposed rats. The exposed organs including brain, and testis gained more weight and their stress parameters [reactive oxygen species (ROS) and thiobarbituric acid reactive substances (TBARS)] increased significantly in a dose dependent manner. Antioxidant enzymes including catalase (CAT), peroxidase (POD), reduced glutathione (GSH) and superoxide dismutase (SOD) reduced significantly in studied organs as compared to control ones. The treated rats have shown atrophy of neurons, microgliosis, cytoplasmic vacuolization, and congestion. Changes in the testis include inflammation, sloughing of cells, damaged spermatogonia, necrosis of spermatids, spermatogonia and arrest of spermatogenesis process. Conclusively, it is suggested that persistent application of nanomaterials at environmentally relevant concentrations may induce adverse toxicological effects in targeted and non-targeted exposed animals.

Keywords: Albino rats, MgO NPs, Oxidative stress, Antioxidant enzymes, Histopathology

Yasir Mahmood1, Nabeel Ijaz2, Aliza Maheen1, Ghulam Mustafa1, Duaa Abdullah Bafail3, Muhammad Rafi Qamar4, Muhammad Aitazaz Ahsan5, Nasir Masood6, Nisreen Rajeh7, Mudassar Mohiuddin8*
1Department of Zoology, The Islamia University, Bahawalpur, Pakistan
2Department of Clinical Sciences, Faculty of Veterinary Sciences, Bahauddin Zakariya University, Multan, Pakistan
3Department of Clinical Pharmacology, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia
4Department of Clinical Medicine and Surgery, Faculty of Veterinary & Animal Sciences, The Islamia University, Bahawalpur, Pakistan
5Department of Pathology, Faculty of Veterinary & Animal Sciences, The Islamia University, Bahawalpur, Pakistan
6Department of Biosciences, COMSATS University Islamabad, Islamabad Campus, Pakistan
7Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah, Saudi Arabia
8Department of Microbiology, Faculty of Veterinary & Animal Sciences, The Islamia University, Bahawalpur, Pakistan

Abstract
ZnO Nanoparticles (ZnO NPs) have wide applications in many fields of life ranging from health, food, agriculture, veterinary medicine, biotechnology, public health, textile and cosmetics. However, exposure to these NPs poses risks to public health, non-target living organisms and the environment. Hence, this study assessed toxicological impacts of ZnO NPs on hematopoietic tissues (bone marrow) and different visceral organs like lungs, intestine and muscles of male Wistar albino rats. Twenty male (20) Wistar albino rats were placed in four groups such as T0 (control group), T1 (50 mg/kg/day ZnO NPs), T2 (75 mg/kg/day ZnO NPs), and T3 (100 mg/kg ZnO NPs). Treated rats exhibited different signs of toxicity like depression and anxiety at higher doses of ZnO NPs. The bone marrow and other visceral organs/tissues were removed and analyzed to know the status of oxidative stress and antioxidant biomarkers. Results revealed notable increase (P≤0.05) in contents of oxidative stress biomarkers (ROS and TBARS) and significant decrease (P≤0.05) in antioxidant enzymes (POD, SOD, CAT, and GSH) in bone marrow as well as lungs, intestine, and muscles (gums) tissues. Histopathological examination indicated degeneration of muscle fibers, atrophied cells and presence of inflammatory materials in muscles (gums) of treated rats. Histologically, lungs were edematous, hemorrhagic and showed severe interstitial pneumonia while necrosis of epithelium of villi along with degeneration of villi in intestine of rats were observed at higher doses of nanoparticles. In conclusion, it can be suggested that ZnO-NPs may induce oxidative stress in multiple visceral organs of albino rats at higher concentrations highlighting disruption of physiological mechanisms.

Keywords: Nanoparticles, Zinc oxide, Nanoparticle toxicity, Oxidative stress biomarkers

Hira Ahsan^1,2, Maria Ayub¹, Mehraj Gul³, Amber Qureshi¹, Hani Z. Asfour⁴, Hafiz Muhammad Bilal⁵, Muhammad Azeem¹, Ammara Wahid¹, Nadeem Ali⁶, Rasheeha Naveed⁷, Mudasar Shabir⁸, Nisreen Rajeh⁹, Ayaz Mammadov¹⁰, Abu Baker Siddique^1*

¹Institute of Microbiology, Government College University, Faisalabad, Pakistan

²Academy of Medical Sciences, College of Henan Medicine, Zhengzhou University, Zhengzhou, Henan, 450052, China

³Quality Control Laboratory, National Institute of Health, Islamabad, Pakistan

⁴Department of Medical Microbiology and Parasitology, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia

⁵Department of Rehabilitation and Allied Health sciences, Riphah International University, Lahore Campus, Lahore, Pakistan

⁶Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah-21589, Saudi Arabia

⁷Institute of Microbiology, University of Agriculture, Faisalabad, Pakistan

⁸Institute of Advanced Materials and Flexible Electronics (IAMFE), Nanjing University of Information Science and Technology, Nanjing, China

⁹Department of Clinical Anatomy, Faculty of Medicine, King Abdulaziz University, Jeddah-21589, Saudi Arabia

¹⁰Department of Life Sciences, Western Caspian University, Baku, Azerbaijan

Abstract

Multi-drug resistant (MDR) bacterial infections rapidly increase morbidity, mortality, and treatment options. Therefore, the search for, development of, or discovery of antimicrobial drugs capable of combating MDR bacteria is urgently needed. The potential of nanotechnology to advance nanomedicine for human health is being studied. The purpose of the present research is to investigate the antimicrobial activity of silver oxide nanoparticles against carbapenem-resistant Pseudomonas aeruginosa and methicillin-resistant Staphylococcus aureus. For this purpose, a total of 240 pus and wound samples were collected from the burn patients and further processed for isolation and identification of P. aeruginosa and MRSA according to standard microbiological techniques. Using a species-specific primer for each bacterial strain, polymerase chain reaction was used for molecular detection. Antimicrobial susceptibility was performed according to the Kirby-Bauer Disk Diffusion method. Molecular detection of carbapenemase-producing P. aeruginosa was performed by PCR by using specific primers. The agar well diffusion assay was used to examine the antibacterial properties of silver oxide nanoparticles, and the broth dilution assay was used to estimate the minimum inhibitory concentration and bactericidal concentration respectively. Out of 240 samples, 42 (17%) were identified as P. aeruginosa and 32 were confirmed as S. aureus isolates. From positive isolates of P. aeruginosa, 25 (59%) were recorded MDR P. aeruginosa and from positive isolates of S. aureus, 18 (56.25%) were detected as MRSA. The highly resistant drug against S. aureus was Penicillin G (100%) followed by Gentamicin (84.37%) and Ciprofloxacin (81.25%). The highly resistant drug against P. aeruginosa was Meropenem (100%), Imipenem (100%) followed by piperacillin (71.42%), gentamicin (64.28%), and ciprofloxacin (64.28%). Out of 42 P. aeruginosa isolates, 8(19%) the prevalence of carbapenemase encoding was noted as blaOXA 3(37.5%), blaNDM 2(25%), blaVIM 1(12.5%) blaKPC 1(12.5%) and blaIMP 1(12.5%).  Silver oxide nanoparticles were considered an effective antibacterial agent with 0.0065mg/mL-0.026mg/mL concentrations that highly inhibited the growth of MRSA and 0.39mg/mL-1.56mg/mL concentrations inhibited the growth of P. aeruginosa. The statistical analysis showed that the MIC and MBC for MDR P. aeruginosa were 0.96±0.43 μg/mL and 1.99±0.90 μg/mL, respectively, while for MRSA they were 0.01±0.008 μg/mL and 0.04±0.012 μg/mL, respectively. The MBC values were higher than MIC values for both pathogens. Silver oxide nanoparticles have such effective antibacterial properties that they can be used as an adequate source of antibacterial agents as alternatives to antibiotics.

Keywords: Silver oxide nanoparticles, Methicillin resistant, Gentamicin, Carbapenemase, Antimicrobial activity

Jingyu Feng^1,2, Li Yang¹, Jiguo Wang^1,2, Jing Zhang^1,2, Lizhu Lin^2,3*

¹Department of Oncology, Shenzhen Bao’an Traditional Chinese Medicine Hospital, Shenzhen 518133, China

²The First School of Clinical Medicine, Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, Guangdong, China

³Department of Oncology, The First Affiliated Hospital of Guangzhou University of Traditional Chinese Medicine, Guangzhou 510405, Guangdong, China

Abstract

The objective of this study was to elucidate the mechanism of the Hexuetongbi formula in treating Chemotherapy-induced Peripheral Neuropathy (CIPN) using network pharmacology, pharmacodynamics, and mechanistic approaches in an oxaliplatin-induced peripheral neuropathy rat model. Network pharmacology is crucial for understanding the multi-target effects of the Hexuetongbi formula on CIPN. This approach allows for a comprehensive mapping of the complex interactions between the formula’s constituents and the biological pathways involved in CIPN, revealing potential synergistic effects and enhancing the formula’s pharmacological validation. The Hexue Tongbi formula’s impact was analyzed on rats undergoing peripheral neuropathy, observing changes in the morphology of L4-6 dorsal root ganglion neurons through hematoxylin and eosin (HE) staining. Simultaneously, a network pharmacology approach was employed, utilizing TCMSP and GeneCards databases to identify common targets between CIPN and Hexue Tongbi’s therapeutic entities. These core targets were scrutinized through GO enrichment and KEGG pathway analysis. To validate the findings, mRNA and protein expression levels in the L4-6 dorsal root ganglion were examined using quantitative PCR and Western Blot assays. Significant modifications were observed in the frequency of cold stimulus withdrawal reflexes and the L4-6 dorsal root ganglion neurons, while the mechanical withdrawal reflex threshold displayed a considerable decrease. In rats treated with the Hexuetongbi formula post-oxaliplatin, there was noteworthy mitigation in the cold stimulation paw withdrawal threshold and augmentation in the mechanical stimulation paw withdrawal threshold. Network pharmacology identified 19 active constituents in Hexuetongbi and 35 targets for CIPN, with the PI3K/Akt signaling pathway emerging as a prominent target. There was a significant upregulation in PI3K, Akt1, Akt2, and Bcl-2 compared to controls, suggesting that Hexuetongbi effectively mitigates oxaliplatin-induced peripheral neuropathy through the modulation of the PI3K/Akt and Bcl-2 pathways.

Keywords: Oxaliplatin, Chemotherapy-induced peripheral neuropathy, Traditional Chinese medicine, Dorsal root ganglion, Apoptosis

Xinyou Liu^2,3, Yihong Luo¹, Zhenglin Wang¹, Cong Wang^1,2,3*

¹Department of General Surgery, Zhongshan Hospital, Fudan University, Shanghai, 200032, China

²Department of General Surgery, Zhongshan Hospital (Xiamen), Fudan University, Xiamen 361015, China

³Xiamen Clinical Research Center for Cancer Therapy, Xiamen 361015, China

Abstract

This study addresses a critical knowledge gap in understanding the tumor microenvironment of thyroid cancer by elucidating the mechanism by which tumor cell-derived CD133-positive exosomes promote lymph node metastasis. We employed molecular docking, western blot, and other molecular characterizations to investigate the crucial interaction between CD133 and VEGFR3 and its impact on metastasis. Additionally, experiments assessed the influence of CD133-positive exosomes on lymphatic endothelial cell proliferation and migration.

Our findings demonstrate a significant direct interaction between CD133 and VEGFR3, as suggested by molecular docking. Furthermore, inhibition of CD133 expression resulted in a notable reduction in lymph node metastasis. We also observed that CD133-positive exosomes derived from thyroid cancer cells actively contribute to the migration and proliferation of lymphatic endothelial cells.

These results unveil a novel pathway for lymph node metastasis in thyroid cancer. The identification of CD133 and its interaction with VEGFR3 as key players in this process holds significant promise for the development of targeted therapeutic strategies.  By focusing on these targets, researchers can potentially improve the prognosis of patients with thyroid cancer.

Keywords: Thyroid cancer cells, Exomes, Lymph node metastasis, CD133, VEGFR3

Edith L. Kadege^1*, Pavithravani B. Venkataramana¹, Teshale Assefa², Joseph C. Ndunguru³, Jean Claude Rubyogo⁴, Ernest R. Mbega¹

¹School of Life Sciences and Bioengineering, The Nelson Mandela African Institution of Science and Technology, Arusha 447, Tanzania

²Crops for Nutrition and Health, International Center for Tropical Agriculture (CIAT), 2704 Arusha, Tanzania

³Department of Research and Innovation, Tanzania Agricultural Research Institute (TARI), Dodoma 1571, Tanzania
⁴Crops for Nutrition and Health, International Center for Tropical Agriculture (CIAT), Nairobi 823-00621, Kenya

Abstract

Tanzania is the top bean producer in Africa and seventh globally, exporting half of its beans to neighboring nations. This study determines common bean varietal performance in northern Tanzania. Disease infection and yield components of 22 genotypes were evaluated in on-station and on-farm trials. The study used a completely randomized factorial trial design with three replications to explore the individual and combined effects of genotype and environment on disease infection and grain yield in two on-station and six on-farm environments. Data were collected on number of emerging plants; canopy height, canopy width, plant vigor, disease infection levels, plant stands at harvest, number of pods per plant, number of grains per pod, 100-grain weight and grain yield and analyzed using R software. The combined analysis of variance revealed significant differences among genotypes, environment, and genotype by environment interactions. Bean canopy height, canopy width, plant vigor and grain yield were high at on-station trials, compared with on-farm trials. Advanced breeding lines showed 56% higher grain yield than commercial checks across study locations. Additive main effects and multiplicative interaction (AMMI) analysis revealed that, genotype was the dominant factor affecting common bean grain yields at 50.3%, whereas the environmental impacts were 25.7%. NUA 48 and NUA 64 were ideal genotypes showing anthracnose resistance and delivering higher grain yield. VTT 923-23-10 and Sweet Violet varieties were stable across the mega-environment. Therefore, NUA 48, NUA 64, VTT 923-23-10 and Sweet Violet are proposed for further evaluation within Tanzanian bean agroecosystem to identify farmers’ preferred varieties.

Keywords: Disease resistance, Yield components, Breeding lines, Bean, Tanzania

Sara Bano¹, Muhammad Imtiaz Rashid², Amna Akhtar¹, Farhan Hafeez¹, Rashid Nazir¹, Faridullah¹, Muhammad Irshad¹, Gabrijel Ondrasek^3*, Akhtar Iqbal^1*

¹Department of Environmental Sciences, COMSATS University Islamabad (CUI), Abbottabad Campus, Tobe Camp, University Road, Abbottabad, Pakistan.

²Center of Excellence in Environmental Studies, King Abdulaziz University, Jeddah 21589, Saudi Arabia.

³Department of Soil Amelioration, Faculty of Agriculture, University of Zagreb, HR-10000 Zagreb, Croatia.

Abstract

To enhance soil fertility and subsequent crop yields, digestate, byproduct of anaerobic digestion, can serve as a supplement or potential alternative to chemical fertilizers when sensibly utilized. This study assessed the impact of two types of digestates on two distinct soils for the perspective of agriculture (affecting pH, EC, organic carbon and mineral nitrogen) and environment (mineral nitrogen leaching under two rainfall patterns). Both soils mainly differed in silt and sand contents: 32% silt and 47% sand for soil-1 and 42% silt and 39% sand for soil-2. Two sets of controlled experiments served the purpose, in which first set involved a soil incubation experiment, applying two digestates to two soil types at 28°C for 60 days. The second set comprised reconstituted soil columns to collect soil solutions at depths of 2.5 cm and 7.5 cm after rainfall application under two patterns for 45 days. Results indicated that application of both digestates in test soils initially increased pH at day 15, followed by a decrease at days 30 and 60. Both digestates significantly elevated soil electrical conductivity compared to control treatments in both soils. Organic carbon content displayed variable impacts, with a slight decrease for solid digestate (12%) and higher decrease for liquid-amended soil (43%) for soil-1. While a significant decrease was observed for soil-2 throughout the incubation period for both amendments (34% and 36% for solid and liquid amended soils respectively). Rapid nitrification occurred with the application of both digestates in both soils, albeit at different rates. Soil-2 exhibited 1.2 to 2 folds higher net nitrification rate (depending upon digestate type and days of incubation) compared to soil-1. Liquid digestate induced more mineral nitrogen compared to solid digestate in both soils. Interestingly, rainfall frequency, digestate type, and soil type influenced the leaching of ammonium and nitrates, with nitrates recording higher levels in both soils, at both depths, and under both rainfall patterns.

 Keywords: Slurry, Nitrates, Crop productivity, Nitrification rate, Soil nutrition