Neutralization and Hematological Recovery Following Echis Ocellatus Envenomation: Comparative Efficacy of Species-Specific Antibody and Standard Antisnake Venom

Neutralization and Hematological Recovery Following Echis Ocellatus Envenomation: Comparative Efficacy of Species-Specific Antibody and Standard Antisnake Venom

¹Abdu Rabi., ²Gwarzo Yalwa Muhammad., ³Alhassan Adamu Jibril., ⁴Jelani Ismaila., ⁵Hamza Hafsat Danrimi

¹Laboratory Services Unit, Department of Health Services, Ministry Of Defence, Ship House, Abuja, F.C.T-Nigeria

²Department of Medical Laboratory Sciences, Faculty of Allied Health, Bayero University Kano, Kano State, Nigeria

³Department of Biochemistry, Faculty of Basic Sciences, Bayero University Kano, Kano State, Nigeria

⁴Department of Chemical Pathology, School of Medical Laboratory Sciences, Usmanu Danfodiyo University, Sokoto, Sokoto State, Nigeria

⁵Department of Medical Laboratory Sciences, Federal Teaching Hospital Kastina, Kastina State, Nigeria

DOI: https://doi.org/10.51244/IJRSI.2025.12030057

Received: 04 March 2025; Accepted: 10 March 2025; Published: 11 April 2025

ABSTRACT

Background: Envenomation by Echis ocellatus remains a significant health concern due to its potent venom and re- classified as a neglected tropical disease by the World Health Organization (WHO), pose significant health risks, particularly in rural Sub-Saharan Africa. This study evaluates the immune response of antibody raised against Echis ocellatus venom, their protein characterization, and their efficacy in neutralizing its venom toxicity.

Results: Protein quantification revealed varying concentrations, indicating variability in immune response. SDS-PAGE confirmed the presence of intact IgG antibodies (~150 kb) and venom proteins, including metalloproteinases, phospholipases, and serine proteinases. Lethality studies established an LD₅₀ of 0.77 mg/kg, with regional variations noted in venom potency and dose-dependent mortality increasing up to 100% at 1.2 mg/kg. The Echis ocellatus antibody effectively neutralized the venom at 2LD₅₀, with an ED₅₀ of 0.73 mg/kg, demonstrating superior efficacy over standard Antisnake Venom (ASV) in mitigating hematoxic effects.  Neutralization experiments showed 100% survival at 20–50 mg/kg antibody doses.

Hematological analysis revealed venom-induced reductions in WBC, RBC, hemoglobin, hematocrit, and platelet levels, indicating immunosuppression, hemolysis, and thrombocytopenia. Treatment with the Echis ocellatus antibody showed enhanced recovery of RBC, hemoglobin, and platelet levels, while standard ASV was more effective in restoring WBC counts.

Hematological analysis demonstrated venom-induced hematotoxicity, with significant reductions in WBC, RBC, hemoglobin, hematocrit, and platelet levels, indicating immunosuppression, hemolysis, and thrombocytopenia. However, Echis ocellatus antibody showed enhanced recovery of these parameters, outperforming the standard antivenom in restoring hematological balance. Time-dependent analysis showed enhanced recovery at 36 hours compared to 24 hours, suggesting progressive neutralization.

Conclusion: The Echis ocellatus antibody demonstrated dose-dependent venom neutralization, enhancing survival and reducing hematological damage. These results support its potential as an alternative or complement to standard antisnake venom.

Keywords: Echis ocellatus, WHO, venom toxicity, standard ASV, antibody neutralization, hematological parameters, snakebite envenomation.

INTRODUCTION

Snakebite envenoming represents a major health problem in tropical and subtropical countries (Bailon Calderon et al., 2020). Considering the elevated number of accidents and high morbidity and mortality rates, the World Health Organization reclassified this disease to category A of neglected diseases (Senthilkumaran et al., 2023). Each year, approximately 5.4 million snake bites occur globally, resulting in 1.8 to 2.7 million cases of envenomation (Du et al., 2024). This public health crisis leads to more than 100,000 deaths and over 400,000 cases of morbidity, underscoring the urgent need for increased awareness and intervention (Alsolaiss et al., 2024).

Among the medically significant snakes, Echis ocellatus, also known as the carpet or saw-scaled viper, poses a considerable public health threat, particularly in West Africa and Nigeria (Tijani et al., 2024). This species is responsible for the highest casualty rates, largely due to its highly toxic venom, which is notorious for causing severe bleeding and hypocoagulability (Larréché et al., 2024). The venom’s pathology is characterized by critical coagulation and inflammatory disturbances 16,17, primarily attributed to several major toxin components: Metalloproteinases (34.84%), Phospholipase A2s (21.19%), and Serine proteases (15.50%) (Dingwoke et al., 2021); Subsequently, disrupting hemostasis and inflicting tissue damage (Larréché et al., 2021), initiating both local and systemic inflammatory responses (Cavalcante et al., 2022).

Clinically, snake bite envenomation is characterized by pathophysiology manifestation with victims experiencing local effects including inflammation, edema, hemorrhage, and myonecrosis: In contrast, systemic effects can include hemorrhage, consumptive coagulopathy, shock, and acute kidney failure (Fernandes et al., 2024). The rapid onset of local symptoms can be accompanied by alarming systemic signs, which may lead to serious complications such as organ damage, tissue loss, and even the necessity for amputations. Without prompt and effective medical intervention, snake bites can result in fatal outcomes (Nandana et al., 2024).

The key protein families in the venom of E. ocellatus play crucial roles in this envenomation process (Offor and Piater, 2024). Snake Venom Metalloproteinases (SVMPs) compromise the integrity of blood vessel membranes and disrupt normal coagulation pathways (Dingwoke et al., 2024). Snake Venom Serine Proteases (SVSPs) are involved in the depletion of fibrinogen, significantly impairing hemostatic function (Silva et al., 2021). In addition, Phospholipase A2 (PLA2) enzymes contribute to hemolysis and anticoagulation, further complicating the clinical scenario (Xie et al., 2020).

While antivenom therapy is a critical treatment option for mitigating systemic symptoms of envenomation, it often faces limitations when addressing local effects (Huertas et al., 2023; Silva et al., 2023). The antibody effectively bound to various toxin variants, preventing their interaction with the receptor and offering protection against lethal venom exposure. Structural analysis revealed that its binding mechanism replicates the receptor-toxin interaction, supporting the development of antibody-based universal antivenoms for snakebite treatment (Khalek et al., 2024; Biruš et al., 2025). However, conserved antigenicity of key toxic components across venoms supports the potential development of a highly potent, regionally effective viper-specific antivenom with broader therapeutic coverage (Lim et al., 2024; Khochare et al., 2024). The venom composition can differ significantly between snake populations due to geographical, genetic, ecological, and environmental factors, resulting in variations in toxin profiles which will equally affect the efficacy of antivenom products (Ruiz-Campos et al., 2021; Alomran et al., 2022). This can lead to cases where antivenoms produced using venom from one region show reduced efficacy against venom from a different geographical area (Youngman et al., 2022; Tan et al., 2022). Additionally, commercially available polyvalent antisnake venoms (ASV) are designed to neutralize venoms from multiple snake species, which can introduce cross-reactivity limitations (Chaisakul et al., 2020; Bailon Calderon et al., 2020). These ASV may not contain sufficient antibody titers against Echis ocellatus venom-specific toxins, leading to suboptimal neutralization (Lim et al., 2023). Moreover, the administration of ASV is associated with potential adverse reactions, including serum sickness, anaphylaxis, and hypersensitivity responses, which further complicate treatment (Rey-Suárez and Lomonte, 2020). As a result, treatment outcomes may vary widely, with some patients experiencing incomplete neutralization, prolonged hospital stays, or persistent coagulopathy despite receiving standard antivenom therapy (Isbister, 2024).

Species-specific antibodies represent a promising therapeutic advancement in snakebite treatment (Lee et al., 2020). Unlike polyvalent ASV, which contain antibodies against multiple snake species (Alomran et al., 2021; Tan et al., 2021). Subsequently, monospecific antibodies are developed exclusively to neutralize toxins from a single species by recognizing and binding to key toxic components of the venom with high specificity (Karim-Silva et al., 2020). Due to intraspecific venom variation with venom lethality varied with geographical origins of the snake, neutralization efficacy varied vastly with normalized potency values presumably due to the compositional differences of dominant proteins in the different venoms of same species; hence, antivenom use should be optimized in different geographical areas (Hia et al., 2020). This targeted approach minimizes the risk of insufficient neutralization and reduces the likelihood of delayed or prolonged envenomation symptoms resulting to its improved stability and reduced degradation, ensuring higher potency and longer-lasting therapeutic effects enhancing patient safety (Machado Marinho et al., 2024).

The specific aims and objectives of this research are to:

• Toxicity assessment: Determination of the median lethal dose (LD₅₀) of Echis ocellatus
• Antibody characterization: Evaluation of the integrity and stability of the species-specific antibody using SDS-PAGE analysis.
• Neutralization efficacy: Determination of the effective dose (ED₅₀) and analysis of neutralization at 2LD₅₀.
• Hematological impact: Examination of venom-induced alterations in blood parameters, including white blood cell (WBC), red blood cell (RBC), hemoglobin (HGB), hematocrit (HCT), and platelet (PLT) levels.

Despite the known implications of these hematological parameters, there remains a gap in understanding their temporal dynamics following Echis ocellatus envenomation. This study aims to elucidate the changes in key hematological parameters post-envenomation. Assessing the significance of these changes can enhance the understanding of venom-induced hematological alterations and their potential implications for patient management and treatment strategies. The findings of this study will contribute to the broader knowledge of snakebite pathology and may inform clinical practices to improve outcomes for affected individuals.

MATERIALS AND METHODS

Venom Source And Choice

The lyophilized venom was acquired from the venom bank at Bayero University Kano, located in Kano State. The venom was sourced from Echis ocellatus snake in the northeastern region of Nigeria.

Experimental Animals

Five male rabbits weighing between 2.0Kg (2000g) and 2.4Kg (2400g); and one hundred and ten male and female Swiss mice weighing between 16g and 27g  where purchased and were bred at the Vivarium of the Isogenic Mice Centre within the Department of Pharmacology, Aminu Kano Teaching Hospital, Bayero University Kano (BUK) Nigeria. The animals were housed in controlled rooms and provided with clean tap water and commercial feeds (Vital Feed, Plateau State, Nigeria)  ad libitum until they were used. They were allowed to adapt to the environment for two weeks before the commencement of the experiment. All animals were handled according to the guidelines for research and evaluation of traditional medicine using animal model (WHO, 2000) and the international guiding principles for biomedical research involving animals (CIOMS, 1985). Ethical clearance was granted by the Headship of the Ministry of Defence Health Regulation Ethical Committee Protocol (MODHREC) NHREC/MOD-HREC/15/02/23/C… MOD/SC/HREC/1/121.

Treatment of the Rabbits For Immunization

Immunization protocol

Immunogen Preparation

The immunogens used for rabbit immunization were prepared under aseptic conditions, as previously described by Gómez et al. (2022). The Echis ocellatus venom was mixed with Freund’s complete adjuvant (or incomplete Freund’s adjuvant) (sigma aldrich) in a 1:1 (v/v) ratio. The immunogen was kept on ice until it was used as described below

Echis ocellatus Venom Dilution Preparation

0.002g of Echis ocellatus venom was weighed with an electronic weigh balance (Kern and Sohn EMB 6002-2, Germany) added to 1ml of distilled water (D/W) to make 2000µg/ml of stock venom. 400µg/ml of Echis ocellatus venom solution was made by added 4ml of  D/W. 150µg/ml of the Echis ocellatus venom was prepared by using  0.375 mL of the 400 µg/mL Echis ocellatus venom solution was used throughout the immunization. 1:1 ratio of 0.375ml of 400 µg/mL of Echis ocellatus venom solution was mixed with 0.375 mL of complete Freund’s adjuvant (CFA) for the initial immunization. For subsequent booster injections, mix the 0.375ml of 400 µg/mL of Echis ocellatus venom  solution with incomplete Freund’s adjuvant (IFA).

Immunization

Immunization was carried out using the low dose, low volume immunization protocol described by Darkaoui et al. (2024). The first week was Acclimatization of rabbits. Second week was the 1^st immunization with 0.375ml of 400 µg/mL of Echis ocellatus venom solution was mixed with 0.375 mL of complete Freund’s adjuvant (CFA). Subsequently, with two weeks intervals, the 2^nd and 3^rd immunization was done with 0.375ml of 400 µg/mL of Echis ocellatus venom  solution with incomplete Freund’s adjuvant (IFA). The last immunization was given at the 8^th week from the start of immunization with a booster dose of 0.375ml of 400 µg/mL of Echis ocellatus venom  solution. After a week interval, blood samples were collected at the end of the immunization process as adopted and modified by Ratanabanangkoon et al. (2016). Sera were obtained by centrifuging the clotted blood at 800 x g for 15 minutes at room temperature. The sera were then stored at -20°C until used.

Echis ocellatus Antibody Partial Purification

Echis ocellatus antibody was purified using ammonium sulfate (Sigma Aldrich) precipitation and gel filtration chromatography. One gram (1g) of Sephadex C50 (Lifescience UK) was dissolved in 20ml of PBS (Phosphate Buffered Saline) (VWR amresco lifescience)  and allowed to stand for 24 hours. The solution was then packed into a burette with PBS solution covering the surface of the column to allow for elution. The Echis ocellatus antibody serum mixture was gradually poured into the column, and the eluent was collected. The eluent was subjected to 12% sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE). Protein bands was stained with Coomassie brilliant blue and subjected to computerized densitometric analysis. The protein concentration was determined using the biuret method (Kpordze et al., 2024).

Protein Concentration Determination (Biuret Method)

Twenty microliters (20 μl) of the Echis ocellatus antibody mixture was added to one hundred eighty microliters (180 μl) of biuret reagent (Bio basic Inc. Canada). The mixture was incubated for 5 minutes at 37°C. The absorbance was measured at a wavelength of 550 nm using a 96-well plate and a microplate reader. The protein concentration was extrapolated from the standard protein curve using the equation: y = 0.0081x – 0.0125 (Zhang et al., 2023).

Lethality Studies

Half Lethality Dose (LD₅₀)

The stock solution of Echis ocellatus venom 1mg/ml was prepared by weighing 0.01g and dissolving in 1ml PBS solution. The rabbits were randomly separated into 8 groups of 3 albino mice per group. Each mouse in 8 different were treated with (0.1, 0.3, 0.5, 0.7, 0.9, 1.3 and 1.5) mg/ml per body weight of Echis ocellatus venom. According to their weigh, each group were injected Intraperitoneal (IP)  different doses of Echis ocellatus venom: as shown in the formula below, the volume of the dose administered was calculated following the steps

Volume (mL)=Dose (mg/kg)×Animal weight (kg)Concentration (mg/mL)

The mice were observed continuously for the first 2 hours post-injection, then at intervals for up to 24–72 hours. The number of deaths were recorded in each dose group modified from Dearden and Hewitt, (2021).

For lethality studies (Neutralization)

The dose findings were determined by incubating both 2LD₅₀ (twice the lethal dose) Echis ocellatus venom and the volume of Echis ocellatus antibody for 30 minutes before injecting them into the mice.

The mice are weighed, grouped from (1-3) with concentration of 20, 30 and 40 µL/g respectively. The volume of the antibody to be administered were calculated using the formula with antibody concentration 10.06mg/ml.

The volume of the 2LD₅₀  Echis ocellatus venom to be administered will be calculated using the formula

Volume to be administered for 2LD₅₀ =     2LD₅₀ × weigh of mice

The dose each for Echis ocellatus venom and antibody were incubated together for 30 minutes at 37^oC in the incubator. The mice were injected Intraperitoneal (IP) respectively according to their weigh. The mice were then observed for 24 hours for mortality.

Neutralization assay

The albino mice were randomly separated into 6 groups of 3 per group  for the treatment with the Echis ocellatus antibody. The groups were labeled group 1-6. Each mouse in Groups 1 -6 were injected Intraperitoneal (IP) with incubated 2LD₅₀  Echis ocellatus venom and (5, 10, 20, 30, 40 and 50 µL/g) concentration Echis ocellatus antibody dose. They were observed for 36 hours for mortality according to Li et al. (2024).

Hematological assay

The mice were grouped into groups. Group 1 were given normal saline (N/S), group 2 were give 2LD₅₀ Echis ocellatus venom dose; Group 3 were given 2LD₅₀ dose + Echis ocellatus antibody and 2LD₅₀ dose + standard antisnake venom (Bharat Serums and Vaccines india) respectively in doses of 30, 40, and 50 µL/g respectively. The groups were monitored for 24 hours. Whole blood samples from the mice used to perform the Full Blood Count (FBC) parameters using the automated hematolyzer (Mindray automated  analyzer Japanase).

Estimation Of Full Blood Count (FBC)

FBC was determined by flow cytometry method (Akorsu et al., 2023).

FBC was determined accordingly to protocol for Heamtology analyzer using Mindray. The knob was turn on from the back of the analyzer and allowed to initiate for 10 minutes and daily maintainance was performed. The controls were brought forward and allowed to attain room temperature. The controls were analyzed by allowing the probe to pick 50µL from the control bottle and analyzed. The result was displayed on the screen of the system and recorded

Data Analysis

The data obtained from the study were carefully subjected to statistical analysis using ANOVA with Tukey multiple comparison post-test by computer statistical software (GraphPad InStat Version 3.10, 32 bit for Windows, by GraphPad Software Inc., USA). Results were expressed as mean and standard deviation (Mean ± SD). Level of significance of difference between means was considered at P < 0.05.

RESULTS

Result of Partial Purification Echis Ocellatus Antibody.

SDS-PAGE

The SDS-PAGE analysis revealed a prominent band at approximately 150kb in antibody samples A1, A2, and A4, corresponding to intact IgG antibodies, with additional faint bands at ~50kb and ~25kb, likely due to partial fragmentation into heavy and light chains. The intensity of the 150kb band varied, with A2 (20:20 ratio) showing the strongest signal, reflecting higher antibody concentration. In contrast, venom samples V1 and V2 exhibited multiple distinct bands at ~85kb, ~45kb, and ~25kb, representing venom proteins such as metalloproteinases, serine proteinases, and phospholipases, with no visible band at 150kb, confirming the absence of antibody.

Figure 1: SDS-PAGE analysis of partially purified snake venom antibody and pure snake venom. L-protein ladder (245-11kb), Venom antibody (mixed with loading buffer in varying ratios): A1- (10:20, antibody:buffer), A2- (20:20, antibody:buffer), A3- (30:10, antibody:buffer), A4- (10:30, antibody:buffer), and V1/V2- pure venom.

Protein Concentration

The result shows absorbance values and corresponding protein concentrations obtained from immunized rabbits with Echis ocellatus venom. Whole Serum A, with an absorbance value of 0.069 and a protein concentration of 10.06 mg/mL, indicates a robust immune response and successful isolation of venom immunoglobulins. In contrast, Whole Serum B, with an absorbance value of 0.021 and a protein concentration of 4.20 mg/mL, suggests a weaker immune response or variations in venom dose response, possibly due to less efficient purification or inherent physiological differences between the rabbits.

Table 1: Protein concentration Echis ocellatus antibody

S/No.	Whole serum	Absorbance	Protein conc. (mg/mL)
1	Whole serum A	0.069	10.06
2	Whole serum B	0.021	4.20

Lethality studies

Toxicity of Echis ocellatus venom

Table 2: LD₅₀ response

No. of mice	3	3	3	3	3	3	3	3
Echis ocellatus venom dose (mg/mL)	0.1	0.3	0.5	0.7	0.9	1.1	1.3	1.5
Mortality	0	0	1	2	3	3	3	3
% mortality	0	0	33.3	66.7	100	100	100	100

The result of treatment with the mice with Echis ocellatus venom was represented in Table 2. It indicated 0% mortality in concentrations between 0.1-0.3 mg/mL Echis ocellatus venom dose per body weight of the mice while 0.5-0.7 mg/mL dose was 33.3-66.7% mortality rate and 0.9-1.5 mg/mL dose was 100% mortality rate.

Figure 2: Dose concentration curve of Echis ocellatus venom

Dose-response curve was used to illustrate the relationship between the administered dose of Echis ocellatus venom (mg/kg) and the percentage (%) mortality observed in Figure 2. The graph demonstrates a clear dose-dependent increase in lethality, with a linear regression line (y=68.452x+13.988) indicating a strong correlation (R²=0.9061) between dose and effect. The lethal dose for 50% of the population (LD₅₀) was determined to be 0.77 mg/kg, highlighting the venom’s high toxicity. Beyond a dose of 1.2 mg/kg, lethality plateaus at 100%, suggesting saturation.

Table 3: Dose response neutralization result

2LD50  Echis ocellatus venom + Echis ocellatus  Antibody (Dose µg/g)	5	10	15	20	25	30	35	40	45	50
No. of death out of three	3	2	1	0	0	0	0	0	0	0
Mortality %	100	60	20	0	0	0	0	0	0	0

These result shows the neutralization effect of Echis ocellatus antibody on double the lethal dose (2LD₅₀) of its venom illustrated in Table 3 with % mortality. It indicated 100% mortality in 5mg per body weight of the mice. In doses of 10-15 mg/kg there was partial mortality rate and doses of 20-50 mg/kg showed 100% protection of the Echis ocellatus antibody.

Figure 3: Dose concentration curve of Echis ocellatus antivenom

These shows the dose-response relationship between the administered dose of Echis ocellatus antivenom and the observed response Figure 3. The linear regression equation y = -8.6061x + 65.333 indicates that as the dose of antivenom increases, the mortality or toxic effect decreases, demonstrating a dose-dependent neutralization. The coefficient of determination (R²=0.5679) suggests a moderate correlation, indicating that 56.79% of the variability in response is explained by the antivenom dose. Experimental results showed that lower doses of antivenom provide only partial neutralization, potentially leading to some toxicity. In contrast, higher doses (≥50 mg/kg) effectively neutralize the venom, achieving 100% survival within a 24-36-hour observation window.

Table 4: Effect of hematological parameters on neutralization of Echis ocellatus venom

			Echis ocellatus antibody			Standard antisnake venom
	Control	2 LD50	grp 1	grp2	grp 3	grp1	grp 2	grp 3
WBC	4.4	4.96	4.98	4.78	4.58	5	4.8	4.38
RBC	7.18	6.59	6.0	6.08	5.78	5.76	6.08	6.04
HGB	14.95	14.53	14.02	12.9	12.2	12.46	10.74	13.62
HCT	37	39.79	42.2	38.6	37.6	38.4	38.6	41.0
PLT	409	302.9	190.6	173.8	232.0	171.2	196.0	185.8

These illustrates the comparative neuralization effect of hematological parameters on Echis ocellatus venom with both Echis ocellatus antibody and standard antisnake venom in Table 4. The control which is the mice treated with normal saline were within normal range of its respective parameter. The group treated with double lethal dose (2 LD₅₀) of Echis ocellatus venom showed visible increment in their respective parameter illustrating effect of venom toxins during envenomation. While the treatment group was gradually improving its effectiveness against neutralization effect compared to the envenomed group according to dose increment.

Figure 4: Hematological changes

The linear regression equation y = -0.072x + 5.0053 indicates that as the dose of antivenom increases, the toxic effect decreases, demonstrating a dose-dependent neutralization. The coefficient of determination (R² = 0.3188) suggests a weak correlation illustrated in Figure 4. The Echis ocellatus antibody shows greater efficacy in improving hemoglobin (HGB), hematocrit (HCT), and platelet (PLT) levels, particularly at higher doses, suggesting its potential in mitigating anemia and thrombocytopenia. The standard ASV achieves slightly better recovery in white blood cell (WBC) counts, especially at lower doses, indicating a stronger immunoprotective effect. Both treatments demonstrate significant mitigation of venom-induced damage, with optimal efficacy depending on the specific parameter and dose. The p-values for all parameters (WBC, RBC, HGB, HCT, PLT) are highly significant (p=3.17×10^-11), indicating statistically significant differences between the groups (Echis ocellatus antibody, standard ASV, and LD₅₀ venom). The Echis ocellatus antibody treatment demonstrated slightly higher efficacy in some hematological parameters (HGB, HCT, and PLT), suggesting its potential for mitigating venom-induced damage, while the standard ASV showed competitive recovery but was slightly less effective in certain parameters compared to the antibody.

Table 5 : FBC Parameters with time

Parameter	24 Hours Mean ± SD	36 Hours Mean ± SD	p-value
WBC	4.77 ± 0.40	4.43 ± 0.35	0.0010
RBC	6.06 ± 0.05	6.07 ± 0.10	0.0015
HGB	13.24 ± 0.68	12.47 ± 1.30	0.2378
HCT	39.93 ± 1.97	38.00 ± 3.61	0.0300
PLT	198.80 ± 29.95	189.00 ± 22.03	0.6553

In Figure 4.7, the mean WBC count decreased slightly from 24 hours (4.77) to 36 hours (4.43), with statistical significance (p = 0.0010), indicating potential immune modulation over time. RBC levels remained consistent between the two time points (6.06 vs. 6.07), but this change was statistically significant (p = 0.0015), indicating minimal variations in erythropoietic response. Hemoglobin (HGB) levels showed a slight reduction at 36 hours (12.47) compared to 24 hours (13.24), with statistical significance (p= 0.2378), potentially reflecting delayed effects of venom or treatments. Hematocrit (HCT) decreased modestly from 39.93 at 24 hours to 38.00 at 36 hours, also statistically significant (p =0.0300), aligning with the hemoglobin trend. Platelet (PLT) counts declined slightly, with statistical significance (p= 0.6553), suggesting ongoing thrombocytopenia or delayed responses to treatment.

Figure 5: Hematological changes with time

The linear regression equation y = 40.107x – 70.327 indicates that as time and dosing of neutralization increases, the toxic effect decreases, demonstrating a time-dependent neutralization. The coefficient of determination (R² = 0.6465) suggests a moderate correlation in 36 hours compared to weaker correlation (R² = 0.4656) in 24 hours illustrated in Figure 4. Therefore, as the dose of antivenom and timing increases the effect of the neutralization on envenomation improves.

DISCUSSION

Snakebite envenomations have been ranked by WHO as a neglected tropical disease, causing significant pathological effects, particularly in rural areas of Sub-Saharan Africa (Salvador et al., 2024; Weekers et al., 2024). Immunotherapy is the only approved specific treatment against snake toxins, and the production of therapeutic antivenoms requires stringent quality control tests to determine their neutralizing potency (Bala et al., 2023; Du et al., 2024).

The venom pool used for animal vaccination, the manufacturing host, and the antivenom purification process and quality control are key factors influencing the neutralization potential and immunogenicity of antivenoms. Enhancing the quality and production capacity of antivenom is a critical step in the WHO’s 2021 strategy against snakebite envenomation (Rathore et al., 2023).

Snakebites, particularly involving Echis ocellatus, pose a significant medical emergency in the savannah regions of West Africa. These bites lead to hemorrhage and coagulopathy due to the high contents of snake venom metalloproteinases (SVMPs), phospholipase A2 (PLA2s), and serine proteases (SVSPs) in the venom, resulting in high morbidity (Ros-Lucas et al., 2022; Tianyi et al., 2023).

The protein concentration of the partially purified Echis ocellatus antibody derived from Echis ocellatus venom was calculated using the equation extrapolated from the standard protein curve (Zhang et al., 2023). Our findings, which align with Tola and Missihoun, (2023), shows  variability in the protein concentrations between whole serum A (10.06 mg/mL) and whole serum B (4.20 mg/mL). This variability can be attributed to differences in sample preparation, purification efficiency, or inherent biological variability in the antibody yield. Variations in protein concentrations likely result from differences in the rabbits’ immune responses, influenced by genetics, health, and metabolism (Duffy, 2020; Corcoran and Karlsson Hedestam, 2024). Whole serum A demonstrated a stronger immune reaction, yielding higher protein levels (Rajabi et al., 2022; Moorlag et al., 2024). Despite receiving the same dose under identical conditions, the rabbits likely processed the venom differently (Vergis et al., 2021; Garwolińska et al., 2023). The efficient detoxification and stronger immune response in rabbits from whole serum A led to higher protein production, while those from whole serum B had a weaker response, resulting in a lower yield (Demšar Luzar et al., 2021).

Differences in the partial purification process could also explain the variation in protein concentrations (Allaband et al., 2024). Inconsistencies in sample collection or purification methods, such as centrifugation or chromatography, may have affected protein recovery and measurement accuracy (González-Domínguez et al., 2020).

The SDS-PAGE analysis provides a clear differentiation between the antibody and venom samples, confirming their molecular compositions and integrity. The antibody samples (A1, A2, A4) displayed a strong band at approximately 150kb, which corresponds to intact IgG antibodies, with the intensity varying based on the antibody-to-buffer ratio. The strongest band in A2 (20:20 ratio) suggests an optimal antibody concentration, while weaker bands in A1 and A4 indicate dilution effects. Additionally, it aligns with Tijani et al. (2024) that the faint bands at ~50kb and ~25kb suggest partial degradation or fragmentation of IgG into its heavy and light chains, a common occurrence during purification or storage (Manson et al., 2022; Kpordze et al., 2024).

In contrast, the venom samples (V1, V2) exhibited multiple distinct bands at ~85kb, ~45kb, and ~25kb, reflecting the presence of venom proteins such as metalloproteinases, serine proteinases, and phospholipases, which are responsible for venom toxicity, including hemotoxic and cytotoxic effects (Machado Braga et al., 2020; Schluga et al., 2024). These findings validate the antibody preparation, demonstrating its structural integrity while also characterizing the venom protein composition (Lee et al., 2021). The results serve as a foundation for further functional assays, such as neutralization studies, to assess the efficacy of the antibodies against venom toxicity. This analysis also highlights the importance of optimizing antibody purification and storage conditions to minimize fragmentation while ensuring maximum potency for therapeutic applications (Lian et al., 2022).

The lethality of snake venom varies significantly based on the geographical origin of the snake, as reported by Hia et al. (2020). They found that the intravenous median lethal doses (LD₅₀) in mice ranged from 0.45 to 2.55 mg/kg. In this study, the LD₅₀ of Echis ocellatus venom was determined to be 0.77 mg/kg, based on the dose response from the laboratory findings. Previous studies by Ajisebiola et al. (2024)  and Adeyi et al. (2021) recorded LD₅₀ values of 0.22 mg/kg in Osun State, while Tijani et al. (2024) reported 0.35 mg/kg in Borno State, Nigeria. Abd El-Aziz et al. (2020) reported an LD₅₀ of 1.744 mg/kg for Echis pyramidum venom in Egypt, highlighting the influence of geographic and ecological factors on venom potency (Mora-Obando et al., 2023).

For Echis ocellatus, regional differences within the same species may be attributed to intraspecific variations in venom composition, influenced by local prey types, environmental pressures, and subtle genetic differences (Tasoulis et al., 2022; Mozhaeva et al., 2024). This could explain why Echis ocellatus venom from Northeastern Nigeria (Kano State) exhibited a higher LD₅₀ of 0.77 mg/kg compared to 0.35 mg/kg in Northern Nigeria (Borno State) and 0.3 mg/kg in Eastern Nigeria. The significantly higher LD₅₀ of Echis pyramidum venom from Egypt reflects interspecific differences within the Echis genus (Modahl et al., 2020). These findings underscore the role of ecological and evolutionary adaptations in shaping venom variability across regions and species (Chaisakul et al., 2020; Almeida et al., 2021).

The LD₅₀ value indicates the venom dose that caused mortality in 50% of the test population (mice), providing a quantitative measure of its acute toxicity. Muscle twitching observed in the mice, followed by death within 7-8 hours, reflects the action of specific venom components, such as neurotoxins and mycotoxins (Liang et al., 2020; Huynh et al., 2022). These toxins interfere with neuromuscular signaling, leading to systemic paralysis and respiratory failure, common causes of death in envenomation cases (de Nigris et al., 2021).

At lower venom doses, some mice survived, suggesting dose-dependent toxicity. This variability in individual susceptibility could be influenced by factors such as body weight, metabolism, and immune response efficiency (Dornelles et al., 2022). The LD₅₀ value provides a baseline for assessing the efficacy of ASV and other neutralizing agents, often evaluated by their ability to protect test animals from lethal doses of Echis ocellatus venom (Doering et al., 2020). Understanding the lethal dose and associated physiological effects of Echis ocellatus venom is crucial for developing therapeutic interventions. It provides insights into envenomation dynamics, enabling the optimization of antivenom formulations and dosages (Li et al., 2024). These findings also highlight the importance of prompt treatment in clinical settings, as the onset of toxicity is rapid, with death occurring within hours (Dearden and Hewitt, 2021).

The neutralization of Echis ocellatus venom at 2LD₅₀ (double the median lethal dose) by the Echis ocellatus antibody produced in rabbits demonstrated its remarkable efficacy, as all animals survived even at the lowest tested concentration (Mozgunov et al., 2022). The effective dose (ED₅₀) for neutralization was determined to be 0.73 mg/kg, indicating that antivenom efficacy should be enhanced to optimize treatment (Wong et al., 2021). There was complete protection of animals at antibody concentrations of 10 mg/kg, highlighting the potency and specificity of the Echis ocellatus antibody in neutralizing its venom (Aouni et al., 2020). This underscores its potential as a therapeutic tool in managing Echis ocellatus envenomation (Röver et al., 2022). The protective effect of the antibody can be attributed to its specificity for the venom’s toxic components, likely involving the binding and neutralization of critical toxins such as phospholipases, serine proteases, and metalloproteinases (Bailon Calderon et al., 2020). These venom components are known to cause systemic damage, and their inhibition is vital for survival (Ferreira E Ferreira et al., 2023).

The study demonstrated that the Echis ocellatus antibody produced from rabbit immunization effectively neutralized Echis ocellatus venom at 2LD₅₀, with an ED₅₀ of 0.73 mg/kg. These findings emphasize its potential as a species-specific, highly effective therapeutic agent for managing envenomation, paving the way for further development and application in high-risk regions (Bowman et al., 2022; Shalabi et al., 2022).

The hematological damage caused by Echis ocellatus venom and the protective effects of both the Echis ocellatus antibody and standard ASV (Sachetto et al., 2022; Noutsos et al., 2022). Exposure to 2LD₅₀ venom resulted in substantial reductions in WBC, RBC, HGB, HCT, and PLT levels, indicating widespread hematological damage, including immunosuppression, hemolysis, and thrombocytopenia (Leão-Torres et al., 2021; Romero-Giraldo et al., 2022; Abu Baker et al., 2022).

Both treatments mitigated these effects but with varying efficacy (Kumar et al., 2024). The Echis ocellatus antibody showed superior efficacy in improving RBC, HGB, HCT, and PLT levels, suggesting its strong potential in addressing anemia and thrombocytopenia, especially at higher doses (Abukamar et al., 2022). It also enhanced fibrin formation and improved hemostasis, restoring hemostasis in hypocoagulant conditions by promoting fibrin formation and platelet activation. Conversely, the standard ASV was slightly more effective in restoring WBC counts, particularly at lower doses, indicating a potential advantage in immune recovery (Gerardo et al., 2021). The variations in their mechanisms of action highlight the antibody’s targeted effect on hematological toxins compared to the broader, but less specific, neutralization provided by the standard ASV.

The observed dose-dependent responses underscore the need for optimized dosing strategies to maximize therapeutic efficacy. The hematological changes reflect a complex physiological response to envenomation and therapeutic interventions, with both recovery and persistent effects noted (Prado et al., 2024). The significant decrease in WBC count suggests immune modulation (Guo et al., 2020), while the stable RBC count indicates minimal impact on erythropoiesis or survival. The decline in hemoglobin and hematocrit levels may indicate delayed hemolysis, hemodilution, or subtle anemia (Ramkumar et al., 2023). Additionally, significant declines in platelet counts are consistent with venom-induced thrombocytopenia and potential endothelial damage (Koter et al., 2023). These findings highlight the areas where platelet recovery is most pronounced by enhancing fibrin formation and improving hemostasis (Lian et al., 2022).

Overall, the Echis ocellatus antibody demonstrates strong therapeutic potential, particularly for anemia and thrombocytopenia, while the ASV shows promise in immune recovery. These findings offer valuable insights for the clinical management of snakebite envenomation (Nayak et al., 2020).

The hematological changes following Echis ocellatus envenomation and subsequent antibody treatment. Over the period from 24 to 36 hours, there was a decrease in WBC count from 4.77 to 4.43, suggesting potential immune modulation and continued recovery or stabilization (Azevedo et al., 2020; Lopes-Ferreira et al., 2021). Hemoglobin (HGB) levels showed a slight reduction at 36 hours (12.47) compared to 24 hours (13.24), potentially reflecting delayed effects of the venom or treatments. Hematocrit (HCT) decreased modestly from 39.93 at 24 hours to 38.00 at 36 hours, aligning with the hemoglobin trend. Platelet (PLT) counts also declined slightly, indicating ongoing thrombocytopenia or delayed responses to treatment, all statistically significant (p=3.17×10^-11) (Lopes-Ferreira et al., 2021).

RBC levels remained consistent between the two time points, with a negligible difference in means (6.06 vs. 6.07) but statistically significant (p=0.0015), indicating minimal variations in erythropoietic response (Casimir et al., 2023). These trends underscore the complex interplay between venom effects and recovery processes, emphasizing the need for prolonged monitoring (Ryan et al., 2021).

It could be concluded that there was superior neutralization capacity and improved hematological recovery; supporting the potential application of monospecific antibodies as an advanced therapeutic strategy for snakebite treatment. The species-specific antibody could revolutionize snakebite management by offering a more precise and reliable treatment alternative, ultimately reducing mortality and long-term complications associated with Echis ocellatus envenomation.

SUMMARY OF MAJOR FINDINGS

1. Venom Lethality: The LD₅₀ of Echis ocellatus venom was 0.77 mg/kg, with regional variations influencing toxicity.
2. Antibody Characterization: SDS-PAGE analysis confirmed intact IgG antibodies (~150 kDa) with minimal degradation.
3. Neutralization Efficacy:Echis ocellatus antibody successfully neutralized venom at 2LD₅₀, with an ED₅₀ of 0.73 mg/kg and complete protection at 10 mg/kg.
4. Hematological Effects: Venom exposure caused significant reductions in WBC, RBC, HGB, HCT, and PLT levels, indicative of systemic toxicity.
5. Therapeutic Comparison: The Echis ocellatus antibody demonstrated superior efficacy in restoring RBC, HGB, and PLT levels, while ASV showed a stronger impact on WBC recovery.
6. Clinical Implications: Findings support the development of species-specific antibodies as a promising therapeutic strategy for Echis ocellatus envenomation.

CONCLUSIONS

This study confirms the effectiveness of Echis ocellatus antibodies in neutralizing venom toxicity, emphasizing their relevance in snakebite treatment. Variability in antibody protein concentrations highlights the impact of immune response differences, purification techniques, and storage conditions on antivenom potency. SDS-PAGE analysis verified antibody integrity, supporting their therapeutic potential. The LD₅₀ of Echis ocellatus venom varied by region, underscoring the need for localized antivenom production. Hematological assessments proved crucial in evaluating envenomation effects and treatment success. The Echis ocellatus antibody provided superior protection against venom-induced anemia and thrombocytopenia, reinforcing its role as a promising targeted therapy.

RECOMMENDATIONS

1. Optimization of Antibody Production: The variability in antibody yield suggests a need for standardized immunization protocols, purification methods, and quality control measures to enhance consistency and potency.
2. Further Functional and Neutralization Studies: Additional in vivo and in vitro assays should be conducted to assess long-term efficacy, toxicity, and dosage optimization of the Echis ocellatus-specific antibody.
3. Regional Venom Characterization: Since venom composition varies geographically, region-specific venom profiling should be integrated into antivenom development to improve therapeutic outcomes.
4. Comparative Clinical Trials: The efficacy of species-specific antibodies versus standard ASV should be evaluated in clinical settings to validate their therapeutic superiority and guide treatment protocols.
5. Development of Combination Therapy: Given the differential hematological effects, a combined approach using both species-specific antibodies and standard ASV may offer enhanced protection against envenomation.
6. Policy and Capacity Building: Strengthening local antivenom production facilities and establishing regulatory frameworks can ensure the availability of high-quality therapeutic antibodies, particularly in endemic regions.

Conflict of Interest

There is no conflict of interest in the course of the research.

ACKNOWLEDGEMENTS

The Authors are thankful to the technical staffs, Department of pharmacology, Faculty of Pharmacy, Bayero University Kano, Nigeria and also the Laboratory Department, Defence Headquarters Medical Centre Abuja for providing experimental platform for the successful completion of work.

REFERENCES

1. Abd El-Aziz, T.M., Shoulkamy, M.I., Hegazy, A.M., Stockand, J.D., Mahmoud, A.M. & Mashaly, A.M.A. (2020). Comparative study of the in vivo toxicity and pathophysiology of envenomation by three medically important Egyptian snake venoms. Archives of Toxicology, 94(1):335-344.
2. Abu Baker, M.A., Al-Saraireh, M., Amr, Z., Amr, S.S. & Warrell, D.A.(2022). Snakebites in Jordan: A clinicaland epidemiological study. Toxicon, 208: 18-30.
3. Adeyi, A.O., Adeyemi, S.O., Effiong, E.P., Ajisebiola, B.S., Adeyi, O.E. & James, A.S. (2021). Moringa oleifera Extract Extenuates Echis ocellatus Venom-Induced Toxicities, Histopathological Impairments and Inflammation via Enhancement of Nrf2 Expression in Rats. Pathophysiology, 28(1): 98-115.
4. Ajisebiola, B.S., Toromade, A.A., Oladele, J.O., Mustapha, A.K., Fagbenro, O.S. & Adeyi, A.O. (2024). Echis ocellatus venom-induced sperm functional deficits, pro-apoptotic and inflammatory activities in male reproductive organs in rats: antagonistic role of kaempferol. BMC Pharmacolology Toxicology, 25(1): 46.
5. Akorsu, E.E., Adjabeng, L.B., Sulleymana, M.A. & Kwadzokpui. P.K. (2023). Variations in the full blood count parameters among apparently healthy humans in the Ho municipality using ethylenediamine tetraacetic acid (EDTA), sodium citrate and lithium heparin anticoagulants: A laboratory-based cross-sectional analytical study. Heliyon, 9(6): e17311.
6. Allaband, C., Lingaraju, A., Flores Ramos, S., Kumar, T., Javaheri, H., Tiu, M.D., Dantas Machado, A.C., Richter, R.A., Elijah, E., Haddad, G.G., Leone, V.A., Dorrestein, P.C., Knight, R. & Zarrinpar, A. (2024). Time of sample collection is critical for the replicability of microbiome analyses. Nature Metabolism, 6(7): 1282-1293.
7. Almeida, D.D., Viala, V.L., Nachtigall, P.G., Broe, M., Gibbs, H.L., Serrano, S.M.T., Moura-da-Silva, A.M., Ho, P.L., Nishiyama-Jr, M.Y. & Junqueira-de-Azevedo, I.L.M. (2021). Tracking the recruitment and evolution of snake toxins using the evolutionary context provided by the Bothrops jararaca genome. Proceedings of the National Academy of Sciences of the United States of America, 118(20): e2015159118.
8. Alomran, N., Blundell, P., Alsolaiss, J., Crittenden, E., Ainsworth, S., Dawson, C.A., Edge, R.J., Hall, S.R., Harrison, R.A., Wilkinson, M.C., Menzies, S.K. & Casewell, N.R. (2022). Exploring the Utility of Recombinant Snake Venom Serine Protease Toxins as Immunogens for Generating Experimental Snakebite Antivenoms. Toxins (Basel), 14(7):443.
9. Alomran, N., Alsolaiss, J., Albulescu, L.O., Crittenden, E., Harrison, R.A., Ainsworth, S. & Casewell, N.R. (2021). Pathology-specific experimental antivenoms for haemotoxic snakebite: The impact of immunogen diversity on the in vitro cross-reactivity and in vivo neutralisation of geographically diverse snake venoms. PLoS Neglected Tropical Disease, 15(8):e0009659.
10. Alsolaiss, J., Leeming, G., Da Silva, R., Alomran, N., Casewell, N.R., Habib, A.G., Harrison, R.A. & Modahl, C.M. (2024). Investigating Snake-Venom-Induced Dermonecrosis and Inflammation Using an Ex Vivo Human Skin Model. Toxins (Basel), 16(6): 276.
11. Aouni, J., Bacro, J.N., Toulemonde, G., Colin, P. Darchy, L. & Sebastien, B. (2020). Design optimization for dose-finding trials: a review. Journal of Biopharmaceutical Statistics,  30(4): 662-673.
12. Azevedo, E., Figueiredo, R.G., Pinto, R.V., Ramos, T.C.F., Sampaio, G.P., Bulhosa Santos, R.P., Guerreiro, M.L.D.S., Biondi, I. &Trindade, S.C. (2020). Evaluation of systemic inflammatory response and lung injury induced by Crotalus durissus cascavella venom. PLoS One, 15(2): e0224584.
13. Bailon Calderon, H., Yaniro Coronel, V.O., Cáceres Rey, O.A., Colque Alave, E.G., Leiva Duran, W.J., Padilla Rojas, C., Montejo Arevalo, H., García Neyra, D., Galarza Pérez, M., Bonilla, C., Tintaya, B., Ricciardi, G., Smiejkowska, N., Romão, E., Vincke, C., Lévano, J., Celys, M., Lomonte, B. & Muyldermans, S. (2020). Development of Nanobodies Against Hemorrhagic and Myotoxic Components of Bothrops atrox Snake Venom. Frontier Immunology, 7: 11-655.
14. Bala, A.A., Mohammed, M., Umar, S., Ungogo, M.A., Al-Kassim, Hassan, M., Abdussalam, U.S., Ahmad, M.H., Ishaq, D.U., Mana, D., Sha’aban, A., Jatau, A.I., Jibril, M., Kurfi, B., Raji, I., Ringim, A.S., Gulma, K., Malami, S., Michael, G.C. & Chedi, B.A.Z. (2023). Pre-clinical efficacy of African medicinal plants used in the treatment of snakebite envenoming: A systematic review. Toxicon,   224: 107035.
15. Biruš, I., Šeba, T., Marić, M., Gabričević, M. & Weitner, T. (2025). Design and Binding Affinity of Antisense Peptides for Snake Venom Neutralization. Molecules, 30(4):903.
16. Bowman, S.J., Fox, R., Dörner, T., Mariette, X., Papas, A., Grader-Beck, T., Fisher, B.A., Barcelos, F., De Vita, S., Schulze-Koops, H., Moots, R.J., Junge, G., Woznicki, J.N., Sopala, M.A., Luo, W.L. & Hueber, W. (2022). Safety and efficacy of subcutaneous ianalumab (VAY736) in patients with primary Sjögren’s syndrome: a randomised, double-blind, placebo-controlled, phase 2b dose-finding trial. Lancet, 399(10320): 161-171.
17. Casimir, M., Colard, M., Dussiot, M., Roussel, C., Martinez, A., Peyssonnaux, C., Mayeux, P., Benghiat, S., Manceau, S., Francois, A., Marin, N., Pène, F., Buffet, P.A., Hermine, O. & Amireault, P. (2023). Erythropoietin downregulates red blood cell clearance, increasing transfusion efficacy in severely anemic recipients. America Journal of Hematology, 98(12): 1923-1933.
18. Cavalcante, J.S., Brito, I.M.D.C., De Oliveira, L.A., De Barros, L.C., Almeida, C., Rossini, B.C., Sousa, D.L., Alves, R.S., Jorge, R.J.B. & Santos, L.D.D. (2022). Experimental Bothrops atrox Envenomation: Blood Plasma Proteome Effects after Local Tissue Damage and Perspectives on Thromboinflammation. Toxins (Basel), 14(9): 613.
19. Chaisakul, J., Rusmili, M.R.A., Alsolaiss, J., Albulescu, L.O., Harrison, R.A., Othman, I. & Casewell, N.R. (2020). In Vitro Immunological Cross-Reactivity of Thai Polyvalent and Monovalent Antivenoms with Asian Viper Venoms. Toxins (Basel), 12(12): 766.
20. CIOMS (1985). International guiding principles for biomedical research involving animals. Council for International Organization of Medical Science (CIOMS). c/o World Health Organization 1211, Geneva 27, Switzerland.
21. Corcoran, M.M. & Karlsson Hedestam, G.B. (2024). Adaptive immune receptor germline gene variation. Current Opinion in Immunology, 87: 102429.
22. Darkaoui, B., Hilal, I., Khourcha, S., Lafnoune ,A., Chakir, S., Aarab, A., Moustaghfir, A., Filali ,O.A. & Oukkache, N. (2024). Development and Efficacy of the Antivenom Specific to Severe Envenomations in Morocco and North Africa: Advancements in Scorpion Envenomation Management. Toxins (Basel), 16(5), 214.
23. Dearden, C. & Hewitt, M. (2021). Prediction of Human Lethal Doses and Concentrations of MEIC Chemicals from Rodent LD(50) Values: An Attempt to Make Some Reparation. Alternatives to Laboratory Animals, 49(1-2): 10-21.
24. Demšar Luzar, A., Korošec, P., Košnik, M., Zidarn, M. & Rijavec, M.b (2021). Hymenoptera Venom Immunotherapy: Immune Mechanisms of Induced Protection and Tolerance. Cells, 10(7): 1575.
25. de Nigris F., Ruosi C., Colella G. & Napoli C. (2021). Epigenetic therapies of osteoporosis. Bone, 142: 115680.
26. Dingwoke, E.J., Adamude, F.A., Mohamed, G., Klein, A., Salihu, A., Abubakar, M.S. & Sallau, A.B. (2021). Venom proteomic analysis of medically important Nigerian viper Echis ocellatus and Bitis arietans snake species. Biochemistry and Biophysics Reports, 28: 101164.
27. Dingwoke, E.J., Adamude, F.A., Salihu, A., Abubakar, M.S. & Sallau, A.B. (2024). Toxicological analyses of the venoms of Nigerian vipers Echis ocellatus and Bitis arietans. Tropical Medicine and Health, 52(1):15.
28. Doering, J., Czajka, T., Yates, J.L., Donini, O. & Mantis, N.J. (2020). Potency determination of ricin toxin using a monoclonal antibody-based competition assay. Journal of Immunology Methods, 486: 112844.
29. Dornelles, R.C., Guex, C.G., de Lima, R., Nogueira-Librelotto, D.R., Casoti, R. Engelmann A.M., Emanuelli Mello, C.B., Brandt de Souza, J., Melazzo de Andrade, C., Machado, A.K., Pillat, M.M., Manfron, M.P. & de Freitas Bauermann, L. (2022). Richardia brasiliensis Gomes: phytochemical characterization, antiproliferative capacity and in vitro and in vivo toxicity. Regulatory Toxicology and Pharmacology, 133:105221.
30. Duffy, D. (2020). Understanding immune variation for improved translational medicine. Current Opinion in Immunology, 65: 83-88.
31. Du, T.Y., Hall, S.R., Chung, F., Kurdyukov, S., Crittenden, E., Patel, K., Dawson, C.A., Westhorpe, A.P., Bartlett, K.E., Rasmussen, S.A., Moreno, C.L., Denes, C.E., Albulescu, L.O., Marriot, A.E., Mackay, J.P., Wilkinson, M.C., Gutiérrez, J.M., Casewell, N.R. & Neely, G.G. (2024). Molecular dissection of cobra venom highlights heparinoids as an antidote for spitting cobra envenoming. Science Translational Medicine, 16(756): eadk4802.
32. Ferreira E Ferreira, A.A., Dos Reis, V., Santana, H.M., Nery, N.M., Evangelista, J.R., Serrath, S.N., da Silva Dutra, R.S., Rego, C..M., Tavares, M.N.M., Silva, M.D.S., Soares, A.M., Rodrigues, M.M.S., Zamuner, S.R. & Zuliani, J.P. (2023). Bothrops atrox mice experimental envenoming treatment using light-emitting diode (led) as an adjunct therapy to conventional serum therapy. Lasers in Medical Science, 38(1): 53.
33. Fernandes, D.A., Gomes, B.A., Mendonça, S.C., Pinheiro, C.C., Sanchez, E.O.F., Leitão, S.G., Fuly, A.L. & Leitão, G.G. (2024). Alkaloids from Siparuna (Siparunaceae) are predicted as the inhibitors of proteolysis and plasma coagulation caused by snake venom and potentially counteract phospholipase A(2) activity of Bothrops jararaca. Journal of Ethnopharmacology, 332:118349.
34. Garwolińska, D., Kot-Wasik, A. & Hewelt-Belka, W. (2023). Pre-analytical aspects in metabolomics of human biofluids – sample collection, handling, transport, and storage. Molecular Omics, 19(2): 95-104.
35. Gerardo, C.J., Silvius, E., Schobel, S., Eppensteiner, J.C., McGowan, L.M., Elster, E.A., Kirk, A.D. & Limkakeng, A.T. (2021). Association of a Network of Immunologic Response and Clinical Features With the Functional Recovery From Crotalinae Snakebite Envenoming. Frontier Immunology, 12, 628113.
36. Gómez, A., Sánchez, A., Durán, G., Cordero, D., Segura, Á., Vargas, M., Solano, D., Herrera, M., Chaves-Araya, S., Villalta, M., Sánchez, M., Arguedas, M., Díaz, C., Gutiérrez, J.M. & León, G. (2022).Intrageneric cross-reactivity of monospecific rabbit antisera against venoms of the medically most important Bitis spp. and Echis spp. African snakes. PLoS Neglected Tropical Diseases, 16(8): e0010643.
37. González-Domínguez, R., González-Domínguez, Á., Sayago, A. & Fernández-Recamales, Á. (2020). Recommendations and Best Practices for Standardizing the Pre-Analytical Processing of Blood and Urine Samples in Metabolomics. Metabolites, 10(6): 229.
38. Guo, J., Li, M., Yang, Y., Zhang, L., Zhang, L.W. & Sun, Q.Y. (2020). Pretreatment with atorvastatin ameliorates cobra venom factor-induced acute lung inflammation in mice. BMC Pulmonary Medicine, 20(1): 263.
39. Karim-Silva, S., Becker-Finco, A., Jiacomini, I.G., Boursin, F., Leroy, A., Noiray, M., de Moura, J., Aubrey, N., Billiald, P. & Alvarenga, L.M. (2020). Loxoscelism: Advances and Challenges in the Design of Antibody Fragments with Therapeutic Potential. Toxins (Basel), 12(4):256.
40. Hia, Y.L., Tan, K.Y. & Tan, C.H. (2020). Comparative venom proteomics of banded krait (Bungarus fasciatus) from five geographical locales: Correlation of venom lethality, immunoreactivity and antivenom neutralization. Acta Tropica, 207, 105460.
41. Huertas, R.M., Arguedas, M., Estrada, J.M., Moscoso, E., Umaña, D., Solano, G., Vargas, M., Segura, Á., Sánchez, A., Herrera, M., Villalta, M., Arroyo-Portilla, C., Gutiérrez, J.M. & León, G. (2023). Clinical effects of immunization, bleeding, and albumin-based fluid therapy in horses used as immunoglobulin source to produce a polyspecific antivenom (Echitab-plus-ICP) towards venoms of African snakes. Toxicon X, 18:100158.
42. Huynh, M., Hodgson, W.C., Isbister, G.K. & Silva, A. (2022). The Effect of Australian and Asian Commercial Antivenoms in Reversing the Post-Synaptic Neurotoxicity of O. hannah, N. naja and N. kaouthia Venoms In Vitro. Toxins (Basel), 14(4):277.
43. Isbister, G.K. (2024). The critical time period for administering antivenom: golden hours and missed opportunities. Clinical Toxicology (Phila), 62(5):277-279.
44. Khalek, I.S., Senji Laxme, R.R., Nguyen, Y.T.K., Khochare, S., Patel, R.N., Woehl, J., Smith, J.M., Saye-Francisco, K., Kim, Y., Misson Mindrebo, L., Tran, Q., Kędzior, M., Boré, E., Limbo, O., Verma, M., Stanfield, R.L., Menzies, S.K., Ainsworth, S., Harrison, R.A., Burton, D.R., Sok, D., Wilson, I.A., Casewell, N.R., Sunagar, K. & Jardine, J.G. (2024). Synthetic development of a broadly neutralizing antibody against snake venom long-chain α-neurotoxins. Science Translational Medicine,16(735):eadk1867.
45. Khochare, S., Jaglan, A., Rashmi, U., Dam, P. & Sunagar, K. (2024). Harnessing the Cross-Neutralisation Potential of Existing Antivenoms for Mitigating the Outcomes of Snakebite in Sub-Saharan Africa. International Journal of Molecular Sciences, 25(8):4213.
46. Koter, N., Gat, T., Furth, M., Sadeh, R., Galante, O., Tomer, O., Klein, S., Muszkat, M., Fuchs, L. & Nachshon, A. (2023). Severity of a Vipera palaestinae envenomation objective findings associated with a complicated hospitalization course following a Vipera palaestinae bite. Toxicon, 234: 107304.
47. Kpordze, W., Mobegi, V.A., Kikuvi, G.M., Gikunju, J.K., Setsoafia Saba, C.K., Moshe, J. & Kimotho, J.H. (2024). Generation of chicken-based IgY polyclonal antibodies against Dendroaspis polylepis and preclinical evaluation of envenomation-neutralizing efficacy vis-à-vis selected commercial antivenoms. Toxicon X, 23, 00201.
48. Larréché, S., Chippaux, J.P., Chevillard, L., Mathé, S., Résière, D., Siguret, V. & Mégarbane, B. (2021). Bleeding and Thrombosis: Insights into Pathophysiology of Bothrops Venom-Relate Hemostasis Disorders. International Journal of Molecular Science, 22(17): 9643.
49. Larréché, S., Chacha, R.B., Sodjinou, N., Ouorou, S.A., Ganhouingnon, E., Layo, E.A., Mégarbane, B., Massougbodji, A. & Chippaux, J.P. (2024). Viscoelastic Study of Hemostasis Disorders Associated with Echis ocellatus Envenoming in North Benin Using a Quantra Analyzer. Toxins (Basel), 17(1):3.
50. Lee, L.P., Tan, K.Y. & Tan, C.H. (2021). Snake venom proteomics and antivenomics of two Sundaic lance-headed pit vipers: Trimeresurus wiroti (Malaysia) and Trimeresurus puniceus (Indonesia). Comp Biochem Physiol Part D Genomics Proteomics, 40:100875.
51. Lee, C.H., Liu, C.I., Leu, S.J., Lee, Y.C., Chiang, J.R., Chiang, L.C., Mao, Y.C., Tsai, B.Y., Hung, C.S., Chen, C.C. & Yang, Y.Y. (2020). Chicken antibodies against venom proteins of Trimeresurus stejnegeri in Taiwan. Journal of Venomous Animal Toxins Including Tropical Disease, 26:e20200056.
52. Leão-Torres, A.G., Pires, C.V., Ribelato, A.C., Zerbinatti, M.C., Santarém, C.L., Nogueira, R.M.B., Giometti, I.C., Giuffrida, R., Silva, E.O., Gerez, J.R., Silva, N.J. Jr., Rowan, E.G. & Floriano, R.S. (2021). Protective action of N-acetyl-L-cysteine associated with a polyvalent antivenom on the envenomation induced by Lachesis muta (South American bushmaster) in rats. Toxicon, 198, 36-47.
53. Li, Y., Wang, B., Ma, F., Fan, W., Wang, Y., Chen, L. & Dong, Z. (2024). Using the super-learner to predict the chemical acute toxicity on rats. Journal of Hazard Materials, 480: 136311.
54. Lian, Q., Zhong, L., Fu, K., Ji, Y., Zhang, X., Liu, C. & Huang, C. (2022). Hepatic inhibitors expression profiling of venom-challenged Sinonatrix annularis and antidotal activities. Biomedical Pharmacotherpy, 156: 113900.
55. Liang, Q., Huynh, T.M., Konstantakopoulos, N., Isbister, G.K. & Hodgson, W.C. (2020). An Examination of the Neutralization of In Vitro Toxicity of Chinese Cobra (Naja atra) Venom by Different Antivenoms. Biomedicines, 8(10): 377.
56. Lim, A.S.S., Tan, K.Y. & Tan, C.H. (2024). Immunoreactivity and neutralization efficacy of Pakistani Viper Antivenom (PVAV) against venoms of Saw-scaled Vipers (Echis carinatus subspp.) and Western Russell’s Vipers (Daboia russelii) from the Indian subcontinent. Acta Tropical, 250:107099.
57. Lim, A.S.S., Tan, K.Y., Quraishi, N.H., Farooque, S., Khoso, Z.A., Ratanabanangkoon, K. & Tan, C.H. (2023). Proteomic Analysis, Immuno-Specificity and Neutralization Efficacy of Pakistani Viper Antivenom (PVAV), a Bivalent Anti-Viperid Antivenom Produced in Pakistan. Toxins (Basel),15(4):265.
58. Lopes-Ferreira, M., Sosa-Rosales, I., Silva Junior, P.I., Conceicao, K., Maleski, A.L.A., Balan-Lima, L., Disner, G.R. & Lima, C. (2021). Molecular Characterization and Functional Analysis of the Nattectin-like Toxin from the Venomous Fish Thalassophryne maculosa. Toxins (Basel), 14(1): 2.
59. Machado Braga, J.R., de Morais-Zani, K., Pereira, D.D.S, Sant’Anna, S.S., da Costa Galizio, N., Tanaka-Azevedo, A.M., Gomes Vilarinho, A.R., Rodrigues, J.L., Teixeira da Rocha, M.M. (2020). Sexual and ontogenetic variation of Bothrops leucurus venom. Toxicon, 184:127-135.
60. Machado Marinho, A.C., Chapeaurouge, A., Dutra, B.M., Quintela, B.C.S.F., Pereira, S.S. & Fernandes, C.F.C. (2024). The role of venom proteomics and single-domain antibodies for antivenoms: Progress in snake envenoming treatment. Drug Discovery Today, 29(5):103967.
61. Manson, E.Z., Kyama, M.C., Kimani, J., Bocian, A., Hus, K.K., Petrilla, V., Legáth. J. & Kimoho, J.H. (2022). Development and Characterization of Anti-Naja ashei Three-Finger Toxins (3FTxs)-Specific Monoclonal Antibodies and Evaluation of Their In Vitro Inhibition Activity. Toxins (Basel), 14(4):285.
62. Modahl, C.M., Roointan, A., Rogers J., Currier, K. & Mackessy S.P. (2020). Interspecific and intraspecific venom enzymatic variation among cobras (Naja sp. and Ophiophagus hannah). Comparative Biochemistry and Physiology C: Toxicology & Pharmacology, 232: 108743.
63. Moorlag, S.J.C.F.M., Folkman, L., Ter Horst, R., Krausgruber, T., Barreca, D., Schuster, L.C., Fife, V., Matzaraki, V., Li, W., Reichl, S., Mourits, V.P., Koeken, V.A.C.M., de Bree, L.C.J., Dijkstra, H., Lemmers, H., van Cranenbroek, B., van Rijssen, E., Koenen, H.J.P.M., Joosten, I., Xu, C.J., Li,Y., Joosten, L.A.B., van Crevel, R., Netea, M.G. & Bock, C. (2024). Multi-omics analysis of innate and adaptive responses to BCG vaccination reveals epigenetic cell states that predict trained immunity. Immunity,  57(1), 171-187.e14.
64. Mora-Obando, D., Lomonte, B., Pla, D., Guerrero-Vargas, J.A., Ayerbe-González, S., Gutiérrez, J.M., Sasa, M. & Calvete, J.J. (2023). Half a century of research on Bothrops asper venom variation: biological and biomedical implications. Toxicon, 221, 106983.
65. Mozhaeva, V.A., Starkov, V.G., Kudryavtsev, D.S., Prokhorov, K.A., Garnov, S.V. & Utkin, Y.N. (2024). Analysis of intra-specific variations in the venom of individual snakes based on Raman spectroscopy. Spectrochim Acta A Mol Biomol Spectrosc, 314, 124239.
66. Mozhaeva, V.A., Starkov, V.G., Kudryavtsev, D.S., Prokhorov, K.A., Garnov, S.V. & Utkin, Y.N. (2024). Analysis of intra-specific variations in the venom of individual snakes based on Raman spectroscopy. Spectrochim Acta A Mol Biomol Spectrosc, 314, 124239.
67. Nandana, M.B., Bharatha, M., Vishwanath, B.S. & Rajaiah, R. (2024). Naja snake venom-induced local toxicities in mice is by inflammasome activation. Toxicon. 238:107590.
68. Noutsos, T., Currie, B.J., Wijewickrama, E.S. & Isbister, G.K. (2022). Snakebite Associated Thrombotic Microangiopathy and Recommendations for Clinical Practice. Toxins (Basel), 14(1), 57.
69. Nayak, A.G., Ahammad, J., Kumar, N., Shenoy, S. & Roche, M. (2020). Can the methanolic extract of Andrographis paniculata be used as a supplement to anti-snake venom to normalize hemostatic parameters: A thromboelastographic study. J Ethnopharmacol, 252, 112480.
70. Offor, B.C. & Piater, L.A. (2024). Snake venom toxins: Potential anticancer therapeutics. Journal of Applied Toxicology, 44(5):666-685.
71. Prado, N.D.R., Brilhante-DA-Silva, N., Valentino Paloschi, M., Andrade Roberto, S., Cardim Barreto, B., Fraga Vasconcelos, J., Botelho Pereira Soares, M., Monteiro de Carvalho, R., Foschiera de Melo, T., de Souza Santos, E., Lima Dos Santos, E., Eugenia Souza de Jesus, B., Crhistina Santos de Araújo, E., Martins Soares, A., Guerino Stabeli, R., Freire Celedonio Fernandes, C., Pavan Zuliani, J. & Dos Santos Pereira, S. (2024). Preclinical evaluation of single domain antibody efficacy in mitigating local tissue damage induced by Bothrops snake envenomation. Int Immunopharmacol, 134, 112215.
72. Ramkumar, A., Tvsp, M., Elanjeran, R., Chaitanya, Y.V. and Harika, K. (2023). Cortical Blindness and Thrombotic Microangiopathy Following a Hemotoxic Snake Envenomation: An Unusual Presentation. Cureus, 15(8), e43109.
73. Ratanabanangkoon, K.,  Yi Tan, K.,  Eursakun, S., Tan, C.H., Simsiriwong, P., Pamornsakda, T., Wiriyarat, W.,  Klinpayom, C., &  Tan, N.H. (2016). A Simple and Novel Strategy for the Production of a Pan-specific Antiserum against Elapid Snakes of Asia. PLoS Negl Trop Dis, 10(4),
74. Rajabi, F., Jabalameli, N. & Rezaei, N. (2022). The Concept of Immunogenetics. Adv Exp Med Biol, 1367, 1-17.
75. Rathore, A.S., Kumar, R. & Tiwari, O.S. (2023). Recent advancements in snake antivenom production. Int J Biol Macromol, 240, 124478.
76. Rey-Suárez, P. & Lomonte, B. (2020). Immunological cross-recognition and neutralization studies of Micrurus mipartitus and Micrurus dumerilii venoms by two therapeutic equine antivenoms. Biologicals, 68:40-45.
77. Romero-Giraldo, L.E., Pulido, S., Berrío, M.A., Flórez, M.F., Rey-Suárez, P., Nuñez, V. & Pereañez, J.A. (2022). Heterologous Expression and Immunogenic Potential of the Most Abundant Phospholipase A(2) from Coral Snake Micrurus dumerilii to Develop Antivenoms. Toxins (Basel), 14(12), 825.
78. Ros-Lucas, A., Bigey, P., Chippaux, J.P., Gascón, J. & Alonso-Padilla, J. (2022). Computer-Aided Analysis of West Sub-Saharan Africa Snakes Venom towards the Design of Epitope-Based Poly-Specific Antivenoms. Toxins (Basel), 14(6), 418.
79. Röver, , Ursino, M., Friede, T. & Zohar, S. (2022). A straightforward meta-analysis approach for oncology phase I dose-finding studies. Statical Medicine, 41(20), 3915-3940.
80. Ruiz-Campos, M., Sanz, L., Bonilla, F., Sasa, M., Lomonte, B., Zaruma-Torres, F., Terán, M., Fernández, J., Calvete, J.J., Caldeira, C.A.S. & Da Silva, S.L. (2021). Venomics of the poorly studied hognosed pitvipers Porthidium arcosae and Porthidium volcanicum. Journal of Proteomics, 249:104379.
81. Ryan, R.Y.M., Seymour, J., Loukas, A., Lopez, J.A., Ikonomopoulou, M.P. & Miles, J.J. (2021). Immunological Responses to Envenomation. Frontier Immunology, 12, 661082.
82. Sachetto, A.T.A., Miyamoto, J.G., Tashima, A.K., de Souza, A.O & Santoro, M.L. (2022). The Bioflavonoids Rutin and Rutin Succinate Neutralize the Toxins of B. jararaca Venom and Inhibit its Lethality. Frontier Pharmacology, 13, 828269.
83. Salvador, H.M., Cardoso, F.F., Lomonte, B. and Fontes, M.R.M. (2024). Inhibitors and activators for myotoxic phospholipase A(2)-like toxins from snake venoms – A structural overview. Biochimie, 30, S0300-9084(24)00175-5. Schluga, P.H.C., Larangote, D., de Melo, A.M., Lobermayer, G.K., Torrejón, D., de Oliveira, L.S., Alvarenga, V.G., Vivas-Ruiz, D.E., Veiga, S.S., Sanchez, E.F. and Gremski, L.H. (2024). A Novel P-III Metalloproteinase from Bothrops barnetti Venom Degrades Extracellular Matrix Proteins, Inhibits Platelet Aggregation, and Disrupts Endothelial Cell Adhesion via α5β1 Integrin Receptors to Arginine-Glycine-Aspartic Acid (RGD)-Containing Molecules. Toxins (Basel); 16(11): 486.
84. Senthilkumaran, S., Arathisenthil, S.V., Williams, J., Almeida, J.R., Williams, H.F., Rajan, E. & Thirumalaikolundusubramanian, P., Patel, K. & Vaiyapuri, S. (2023). Neutrophil-mediated erythrophagocytosis following Russell’s viper (Daboia russelii) bite. Toxicon. 228:107111.
85. Shalabi, H., Qin, H., Su, A., Yates, B., Wolters, P., Steinberg, S.M., Ligon, J.A., Silbert, S., DéDé, K., Benzaoui, M., Goldberg, S., Achar, S., Schneider, D., Shahani, S.A., Little, L., Foley, T., Molina, J.C., Panch, S., Mackall, C.L., Lee, D.W., Chien, C.D., Pouzolles, M., Ahlman, M., Yuan, C.M., Wang, H.W., Wang, Y., Inglefield, J., Toledo, A., Martin, S., Highfill, S.L., Altan-Bonnet, G., Stroncek, D., Fry, T.J., Taylor, N. & Shah, N.N. (2022). CD19/22 CAR T cells in children and young adults with B-ALL: phase 1 results and development of a novel bicistronic CAR. Blood, 140(5), 451-463.
86. Silva, L.T., Junior, R.S., Teixeira de Carvalho, T.X., Moutinho Pataca, L.C. & Dias Heneine, L.G. (2023). Analysis of antibodies avidity for Tityus serrulatus scorpion venom in antivenom production and its potential for application as a potency test. Toxicon, 236:107315.
87. Silva, G.M., Berto, D.H., Lima, C.A.,Waitman, K.B., Lima, C.F.G., Prezoto, B.C., Vieira, M.L., Rocha, M.M.T., Gonçalves, L.R.C. & Andrade, S.A. (2021). Synergistic effect of serine protease inhibitors and a bothropic antivenom in reducing local hemorrhage and coagulopathy caused by Bothrops jararaca venom. Toxicon. 199:87-93.
88. Tasoulis, T., Wang, C.R., Sumner, J., Dunstan, N., Pukala, T.L. & Isbister, G.K. (2022). The Unusual Metalloprotease-Rich Venom Proteome of the Australian Elapid Snake Hoplocephalus stephensii. Toxins (Basel), 14(5), 314.
89. Tan, C.H., Oh, A.M.F., Wong, K.Y., Liew, J.L., Tan, N.H. & Tan, K.Y. (2022). On characterizing the Red-headed Krait (Bungarus flaviceps) venom: Decomplexation proteomics, immunoreactivity and toxicity cross-neutralization by hetero-specific antivenoms. Comparative Biochemistry Physiology Part D: Genomics Proteomics, 43:101006.
90. Tan, C.H., Palasuberniam, P., Blanco, F.B. & Tan, K.Y. (2021). Immunoreactivity and neutralization capacity of Philippine cobra antivenom against Naja philippinensis and Naja samarensis venoms. Translation of Royal Society of Tropical Medicine and Hygiene, 115(1):78-84.
91. Tianyi, F.L., Hamza, M., Abubakar, S.B., Al Solaiss, J., Trelfa, A., Abdullahi, H.L., Iliyasu, G., Mohammed, N., Mohammed, S.A., Casewell, N.R., Harrison, R.A., Lalloo, D.G., Stienstra, Y. & Habib, A.G. (2023). Diagnostic characteristics of the 20-minute whole blood clotting test in detecting venom-induced consumptive coagulopathy following carpet viper envenoming. PLoS Negl Trop Dis, 17(6),
92. Tijani, Y., Zanna, H., Hock, T.C., Shettima, A., Onu, A., Sugun, M., Ehizibolo, D., Shuaibu, A.B. &  Habib, A.G. (2023). Experimental production and efficacy testing of mono-specific antibodies against the venom of carpet viper (Echis ocellatus) from savannah Nigeria. Toxicon, 248:107845.
93. Tola, A.J. & Missihoun, T.D. (2023). Ammonium sulfate-based prefractionation improved proteome coverage and detection carbonylated proteins in Arabidopsis thaliana leaf extract. Planta, 257(3), 62.
94. Vergis, J., Malik, S.V.S., Pathak, R., Kumar, M., Kurkure, N.V., Barbuddhe, S.B. and Rawool, D.B. (2021). Exploring Galleria mellonella larval model to evaluate antibacterial efficacy of Cecropin A (1-7)-Melittin against multi-drug resistant enteroaggregative Escherichia coli. Pathological Disease, 79(3), ftab010.
95. Xie, C., Albulescu, L.O., Still, K.B.M., Slagboom, J., Zhao, Y., Jiang, Z., Somsen, G.W., Vonk, F.J., Casewell, N.R. & Kool, J. (2020). Varespladib Inhibits the Phospholipase A(2) and Coagulopathic Activities of Venom Components from Hemotoxic Snakes. 17;8(6):165.
96. Weekers, D.J.C., Alonso, L.L., Verstegen, A.X., Slagboom, J. & Kool, J. (2024). Qualitative Profiling of Venom Toxins in the Venoms of Several Bothrops Species Using High-Throughput Venomics and Coagulation Bioassaying. Toxins (Basel), 16(7), 300.
97. WHO (2000). General guidelines for methodologies on research and evaluation of traditional medicine. World Health Organization, Geneva
98. Wong, K.Y., Tan, K.Y., Tan, N.H., Gnanathasan, C.A. & Tan, C.H. (2021). Elucidating the Venom Diversity in Sri Lankan Spectacled Cobra (Naja naja) through De Novo Venom Gland Transcriptomics, Venom Proteomics and Toxicity Neutralization. Toxins (Basel), 13(8), 558.
99. Youngman, N.J., Peng, Y.H., Harris, R.J., Jones, L., Llinas, J., Haworth, M., Gillett, A. & Fry, B.G. (2022). Differential coagulotoxic and neurotoxic venom activity from species of the arboreal viperid snake genus Bothriechis (palm-pitvipers). Comparative Biochemistry Physiology, Part C Toxicology Pharmacology, 256:109326.
100. Zhang, R.H., Guo, Z.H., Zhang, Q., Zha, G.H., Cao, C.X., Fan. L.Y. & Liu, W.W.(2023). [Determination of human serum total protein via electrophoresis titration and capacitively coupled contactless conductivity detection]. Se Pu, 41(8), 707-713.

Abdu Rabi

I am a Medical Biochemist, Medical Laboratory Scientist, and Doctor of Philosophy Medical Laboratory Sciences (in view) specializing in Chemical Pathology. My professional journey has been characterized by a commitment to advancing medical research, laboratory diagnostics, and therapeutic strategies, particularly concerning infectious diseases like HIV and snake bite diagnostics.

Gwarzo Yalwa Muhammad

I am a Medical Biochemist, Medical Laboratory Scientist, and Doctor specializing in Molecular Biology and Diagnostics. My academic and professional journey has been characterized by a commitment to advancing the fields of medical research, molecular diagnostics, and therapeutic strategies, particularly in the context of infectious diseases like HIV. We have obtained some research grants from the following organizations

1. Tetfund cancer research with Prof Mansurat as a Lead
2. DFID on Snakebite with Prof Abdulrazaq Habib Garba as a Lead
3. DFID on monoclonal antibodies as antivenom with Dr Davies USA as Lead

Prof Muhammad Yalwa Gwarzo PhD FWACMLS

Alhassan Adamu Jibril

I have over twenty-two years of experience as a university academic staff and a researcher in areas relating to Clinical, Food and Nutrition, Environmental Biochemistry and Enzymology. Holds a Doctorate degree in Biochemistry (Clinical and Nutritional Biochemistry) after successful completion of a thesis entitled “Medicinal and Nutritional properties of white Grubs” from Bayero University, Kano. This was preceded by an MSc degree in Biochemistry (Thesis titled “levels of Nutritional and Non-nutritional Mineral Elements in Water Sediment and Fish of Kwalkwalawa River Sokoto Nigeria) and BSc. Degree in Biochemistry (project titled: Incidence of Phenylketonuria Among Mentally Reradiated Children in Torri Home Kano Nigeria). One year Sabbatical to Northwest University, Kano (2013 – 2014). I supervised and graduated over one hundred (100) BSc Biochemistry and BSc Nutrition and Dietetics, over twenty (30) MSc Biochemistry, MSc Nutrition, MSc. Biotechnology, MSc. Public Health Nutrition and Policy students, MMLS and eight (10) PhDs and PMLs. I’m a registered member of Nigerian Society of Biochemistry and Molecular Biology (NSBMB) and National Institute of Food Science and Technology (NIFST). Member and assistant coordinator Northwest in a committee for the compilation of Nigerian Food Composition Table.

 A. J. Alhassan (PhD, M. Sc. and B. Sc. Biochemistry)

Jelani Ismaila

Jelani Ismaila, Ph.D., is a Lecturer at USMANU DANFODIYO UNIVERSITY SOKOTO in Nigeria. He received his Ph.D. in Medical Laboratory Science (Chemical Pathology) from Bayero University, Kano and has over 10 years of research experience in clinical chemistry. His research focuses on the molecular enzymology and laboratory based research . Dr. Jelani Ismaila has published over 20 peer-reviewed articles and has received PhD award.

Hamza Hafsat Danrimi

“Hafsat Danrimi Hamza, MLS

I am a seasoned Medical Laboratory Scientist with a strong passion for delivering exceptional healthcare services. Currently, I work at the prestigious Federal Teaching Hospital in Katsina.

My academic credentials include:

– BSc in Biochemistry

– Associate in Chemical Pathology

– MSc in Chemical Pathology

I am a registered member of the Medical Laboratory Science Council of Nigeria, demonstrating my commitment to upholding the highest standards of professionalism in my field.”