Botanical Anthelmintics for Sustainable Sheep Production: In Vivo Efficacy of Carica papaya Seeds and Moringa oleifera Leaves Against Haemonchus contortus and Moniezia spp. in Ghana

Botanical Anthelmintics for Sustainable Sheep Production: In Vivo Efficacy of Carica papaya Seeds and Moringa oleifera Leaves Against Haemonchus contortus and Moniezia spp. in Ghana | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Botanical Anthelmintics for Sustainable Sheep Production: In Vivo Efficacy of Carica papaya Seeds and Moringa oleifera Leaves Against Haemonchus contortus and Moniezia spp. in Ghana PRINCE KYERE DWAAH, Nana Yaa Awua-Boateng, Sylvia Afriyie Squire, and 4 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-8403819/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Gastrointestinal helminths, particularly Haemonchus contortus (H. contortus) and Moniezia spp., continue to constrain sheep productivity in sub-Saharan Africa. Increasing reports of reduced efficacy of conventional anthelmintics underline the need to evaluate sustainable alternative control options. This study assessed the in vivo anthelmintic efficacy of aqueous extracts of Carica papaya seeds (CPS) and Moringa oleifera leaves (MOL) in naturally infected sheep under field conditions in Ghana. Thirty-three sheep were allocated to four treatment groups: CPS, MOL, conventional dewormer, and distilled water. Treatments were administered orally on Day 0, and faecal egg counts (FEC) were determined on Days 0, 5, 10, and 30 using the McMaster technique. Anthelmintic efficacy was evaluated using the faecal egg count reduction test (FECRT), and body weight changes were monitored over the study period. By Day 30, MOL achieved a mean FECRT of 92.1% against Moniezia spp. and 60.9% against H. contortus , while CPS achieved reductions of 82.4% and 51.3%, respectively. The conventional dewormer showed lower efficacy (≤ 50%) for both parasites. Sheep receiving MOL exhibited the greatest mean body weight gain (4.7 kg), although differences among treatments were not statistically significant. Egg counts increased markedly in the distilled water group. The findings indicate that MOL and CPS exhibit moderate to high anthelmintic activity under field conditions and may offer complementary options for helminth control in sheep production systems where resistance to conventional anthelmintics is of concern, which may contribute to integrated parasite management. Further studies with larger sample sizes and standardized extract characterization are warranted. Sheep Helminths Anthelmintic resistance Ethnoveterinary Medicine Faecal Egg Count Reduction Anthelmintic Efficacy Figures Figure 1 Figure 2 Figure 3 Figure 4 INTRODUCTION Sheep production constitutes an important component of smallholder livestock systems across Africa, contributing to household income, food security, and livelihood resilience. In West Africa, sheep are commonly managed under extensive and semi-intensive systems, where productivity is closely linked to environmental conditions and animal health management practices (Food and Agriculture Organization {FAO}, 2021 ; Thornton & Herrero, 2021). Despite their adaptability, productivity in these systems is frequently constrained by parasitic diseases, with gastrointestinal helminths remaining among the most persistent challenges to sustainable sheep production. Among the helminths affecting small ruminants, Haemonchus contortus ( H. contortus ) and Moniezia spp. are widely distributed in tropical and subtropical regions. H. contortus is regarded as one of the most pathogenic nematodes of sheep due to its hematophagous nature, high fecundity, and capacity for rapid population expansion under favourable climatic conditions, leading to anaemia, reduced growth, and increased mortality (Kaminsky et al. 2022 ; Papadopoulos et al. 2021 ). Although infections with Moniezia spp. are often considered of lower clinical importance, heavy or persistent burdens, particularly in young animals, may compromise feed efficiency and growth performance (Torres-Acosta & Hoste, 2023 ). Control of gastrointestinal helminths has historically depended on the regular use of broad-spectrum anthelmintic drugs. While this approach initially resulted in substantial improvements in livestock productivity, increasing reports of reduced drug efficacy have emerged globally (Vineer et al. 2020 ). Anthelmintic resistance (AR) is now widely recognized as a major constraint to parasite control in small ruminants, particularly in low- and middle-income countries where treatment is often frequent, diagnostics are limited, and dosing practices may be imprecise (Levecke et al. 2022 ; Coles et al. 1992 ). Under such conditions, reliance on a narrow range of conventional anthelmintics has raised concerns about the long-term sustainability of conventional parasite control programmes. In response to this challenge, attention has increasingly turned toward integrated parasite management approaches that combine strategic drug use with complementary control options. Ethnoveterinary medicine, which encompasses the use of locally available plant-based remedies, has gained renewed interest as a potential component of such strategies (Eguale 2021 ). Numerous medicinal plants have been reported to possess antiparasitic properties; however, their efficacy varies widely depending on plant species, preparation method, parasite species, and evaluation protocol (Belga et al. 2024 ; Jato et al. 2022 ). Moreover, many studies rely on in vitro assays or solvent-based extracts, limiting their applicability under field conditions typical of smallholder systems. Moringa oleifera ( M. oleifera ) and Carica papaya ( C. papaya ) are widely distributed in Sub-Saharan regions and commonly used for nutritional and medicinal purposes. Experimental studies suggest that extracts from M. oleifera leaves and C. papaya seeds contain bioactive compounds capable of disrupting helminth development and survival (Páez-León et al. 2022 ; Cabral et al. 2019 ). Nevertheless, in vivo evidence derived from naturally infected sheep, particularly using aqueous preparations representative of practical on-farm use in West Africa, remains limited. The present study therefore, assessed the anthelmintic activity of aqueous extracts of M. oleifera leaves and C. papaya seeds in sheep naturally infected with H. contortus and Moniezia spp. under field conditions in Ghana. By comparing their effects with a commonly used synthetic anthelmintic and an untreated control, the study seeks to generate applied evidence relevant to sustainable parasite control in tropical small ruminant production systems. METHODOLOGY Study Area The study was conducted at the Disease Investigation Farm (DIF) in Techiman, Bono East Region, Ghana, an area known for its high prevalence of gastrointestinal parasitic infections in small ruminants, to investigate conventional dewormers by the Veterinary Directorate Service (VSD) of Ghana (VSD, 2019; MoFA, 2014). The site lies within the transitional ecological zone (7.59529° N, -1.93331° W), characterized by Guinea savannah and semi-deciduous vegetation. Techiman experiences a tropical savannah climate with annual rainfall of 1,200-1,500 mm and temperatures ranging from 22°C to 34°C during the study period (September-October 2024). Plant Materials and Preparation of Extracts Papaya seeds and moringa leaves were selected based on prior ethnoveterinary evidence and unpublished work by the research team. Mature papaya fruits and fresh moringa leaves were harvested from organically maintained plants at DIF. Plant materials were shade-dried at ambient temperature (25–30°C) for 10–14 days, ground, and sieved (1 mm mesh). Fifty grams of each powdered sample were macerated in 500 mL of distilled water (1:10 w/v) for 12 hours, filtered through sterile muslin, and stored at 4°C for not more than 72 hours, following established protocols (Belga et al. 2024 ; Kedir, 2024 ; Jato et al. 2022 ; Castangna et al. 2021; Bessell et. al. 2021). Determination of Safe Dosage A pilot dose-ranging trial was performed using 1–10 mL/kg BW of each extract, observing for toxicity or behavioural changes over 72 hours. No adverse effects were recorded; therefore, 10 mL/kg BW was selected as the therapeutic dose for the main trial. Experimental Design A randomized controlled design was used involving 33 naturally infected sheep. Sample size determination followed the statistical framework of (Levecke et al. 2022 ; Coles et al. 1992 ). Animals were randomly allocated into four treatment groups as shown in Table 1 Table 1 Experimental groups, treatments, and administered dosages in naturally infected sheep. Group Treatments Dosage 1 Distilled water (negative control) 10 mL/kg BW 2 Papaya seed extract 10 mL/kg BW 3 Moringa leaf extract 10 mL/kg BW 4 Albendazole (positive control) 10 mg/kg BW Footnote : BW = body weight. All treatments were administered orally on Day 0. Faecal samples were collected on Days 0, 5, 10, and 30 for FEC analysis. Faecal Collection and Parasitological Examination Faecal samples were obtained directly from the rectum using standard operating procedures (Denwood et. al 2023 ). Quantitative FECs were performed using the McMaster technique (Mohammedsalih et. al, 2025 ; Charles Sturt University, 2024 ; Playford & Besier, 2024 ; Coles et. al, 1992 ). Three grams of faeces were homogenized in 60 mL of saturated NaCl solution, sieved, and examined microscopically (×10 objective) for helminth eggs within the grid area. Laboratory personnel were blinded to treatment allocation to minimize bias. The Faecal Egg Count Reduction Test (FECRT) was adopted from (Denwood et. al 2024; Coles et. al. 1992 ): FECRT (%) = \(\:\frac{\left(Mean\:FEC\:before\:treatment\right)-\left(Mean\:FEC\:after\:treatment\right)}{\left(Mean\:FEC\:before\:treatment\right)}\) ×100, using pre-treatment (Day 0) and post-treatment (Day 30) values for Haemonchus contortus and Moniezia spp. Statistical Analysis Descriptive statistics (mean ± SD) were computed using Microsoft Excel 2019. Data normality was assessed with the Shapiro-Wilk test. One-way ANOVA was used to compare mean FECs among groups, with Tukey’s Honest Significant Difference (HSD) post hoc test for pairwise comparisons (p < 0.05). Two-way ANOVA evaluated treatment × time interaction effects, and paired t-tests compared pre- and post-treatment values within groups. Post hoc power analysis (α = 0.05) confirmed sample adequacy (> 0.8). RESULTS Helminth Load before Treatment At baseline, 60.6% (20/33) of sheep were co-infected with H. contortus and Moniezia spp., while 30.3% (10/33) were infected with H. contortus alone and 9.1% (3/33) with Moniezia spp. alone (Table 2 ). Parasite burdens ranged from moderate to high according to Veterinary Services Directorate classification criteria. Rams showed a higher mean FEC than females and younger animals, although these differences were not subjected to statistical testing due to sample size constraints. There were no significant differences in baseline FECs among treatment groups for either H. contortus (p = 0.72) or Moniezia spp. (p = 0.68), indicating comparability across groups before treatment. Table 2 Pre-treatment distribution of H. contortus and Moniezia spp. infections by sex and age category (n = 33 sheep). Sex and Age No. Sheep Type of infection H. contortus only Moniezia spp . only Both helminths Ram 4 - - 4 Ewe 21 8 2 11 Young Ram 4 2 - 2 Young Ewe 4 - 1 3 n (%) 33 (100) 10 (30.3) 3 (9.1) 20 (60.6) Footnote : Both M. expansa and M. benedeni were detected and are reported collectively as Moniezia spp. FECRT of Aqueous Moringa oleifera Leaves and Carica papaya Seeds against H. contortus Reductions in H. contortus FEC differed among treatment groups over time (Fig. 1). From Fig. 2 , by Day 30 post-treatment, sheep receiving aqueous M. oleifera leaf extract showed a mean FECRT of 60.9% (95% CI: 52.2–69.5), while those treated with aqueous C. papaya seed extract achieved a reduction of 51.3% (95% CI: 43.1–59.4). Conventional dewormer treatment resulted in a mean FECRT of 43.9% (95% CI: 35.4–52.1). In contrast, mean FEC in the untreated control group increased over the same period. One-way ANOVA indicated significant differences in mean FECs between treated groups and the untreated control at Day 30 (p < 0.05). However, when evaluated using two-way ANOVA, neither the main effects of treatment (p = 0.219) nor time (p = 0.117) nor their interaction (p = 0.995) were statistically significant. FECRT of aqueous oleifera leaves and aqueous C. papaya seeds against Moniezia spp. A progressive reduction in Moniezia spp. FEC was observed in sheep treated with both plant extracts (Fig. 3). From Fig. 4 , by Day 10, aqueous M. oleifera leaf extract achieved a FECRT of 52.6% ± 6.7 (95% CI: 40.2–65.1), which increased to 92.1% ± 4.8 (95% CI: 83.6–100.0) by Day 30. Sheep treated with aqueous C. papaya seed extract showed mean FECRTs of 35.3% ± 5.5 (95% CI: 24.1–46.3) at Day 10 and 82.4% ± 6.2 (95% CI: 70.1–94.6) at Day 30. Conventional dewormer treatment resulted in a mean FECRT of 50.0% ± 7.3 (95% CI: 33.5–66.5) by Day 30. In contrast, the untreated control group exhibited a mean increase in egg counts of 111.8% ± 9.2% over the study period. Effect of Treatment Type and Time on Anthelmintic Efficacy Two-way ANOVA was used to assess the effects of treatment, time, and their interaction on FECRT% (Table 3 ). No statistically significant main effects of treatment or time were detected, and no considerable treatment × time interaction was observed (p > 0.05 for all comparisons). These results indicate substantial within-group variability and suggest that treatment-associated changes over time were not consistent across all animals. Exploratory post hoc comparisons suggested within-group reductions in egg counts over time for sheep receiving M. oleifera . Still, these findings should be interpreted cautiously, given the absence of significant overall interaction effects. Table 3 Two-way ANOVA assessing the effects of treatment, time, and their interaction on faecal egg count reduction (%). Source of Variation Sum of Squares df F-Statistic p-Value Treatment Type 2276.78 2 1.8 0.219 Time 3468.11 2 2.75 0.117 Treatment × Time (Interaction) 115.22 4 0.046 0.995 Residual (Error) 5677.5 9 Interpretation Lack of significant interaction (p = 0.995) implies consistent treatment effect over time. Weight Gain Change Mean body weight increased over time in all treated groups (Table 4 ). Sheep treated with aqueous M. oleifera leaf extract exhibited the greatest numerical increase in mean body weight over the 30 days (mean gain: 4.7 kg), followed by those treated with aqueous C. papaya seed extract and Conventional dewormer. Sheep in the untreated control group showed minimal change in body weight over the same period. One-way ANOVA did not detect statistically significant differences in body weight among treatment groups at any sampling point (p > 0.05). Table 4 Mean body weight (kg, mean ± SD) of sheep by treatment group at each sampling time. Treatment Days 0 5 10 30 P-value Conventional Dewormer 24.13 ± 2.5 25.06 ± 2.5 26.13 ± 2.4 27.63 ± 2.4 0.770 Aqueous moringa leaves 24.5 ± 2.9 25.63 ± 3.0 27.06 ± 3.1 29.19 ± 3.1 0.720 Aqueous papaya seeds 26 ± 2.9 27.06 ± 2.8 28.06 ± 2.8 29.06 ± 2.9 0.886 Distilled Water 26 ± 2.6 25.44 ± 2.5 26.22 ± 2.7 26.39 ± 2.8 0.995 Footnote : No significant differences in body weight were detected among treatment groups (one-way ANOVA; p > 0.05). Post Hoc Analysis Although the overall treatment × time interaction was not significant, exploratory pairwise comparisons indicated within-group changes over time, as shown in Table 5 . These findings should be interpreted cautiously, given the absence of statistically significant main effects. Table 5 Tukey’s HSD pairwise comparisons of faecal egg count reduction (%) across sampling days. Group 1 Group 2 Mean Diff p-Value 95% CI (LP) Significant C.D-Day 0 C.D-Day10 30 0.939 -69.36 to 129.36 No C.D-Day 0 C.D-Day5 17 0.998 -82.36 to 116.36 No C.D-Day10 ACPS-Day 10 -20.0 0.994 -119.36 to 79.36 No ACPS-Day 0 AMOL- Day 10 57.5 0.43 -41.86 to 156.86 No ACPS-Day 5 AMOL-Day 5 21 0.992 -78.36 to 120.36 No Footnote : i. CD is Conventional Dewormer, ACPS is Aqueous Carica Papaya seeds, and AMOL is Aqueous Moringa oleifera leaves ii. Pairwise comparisons are exploratory and should be interpreted in the context of the non-significant overall ANOVA. DISCUSSION This study assessed the in vivo efficacy of two ethnoveterinary plant extracts against gastrointestinal helminths in sheep under natural grazing conditions. Both aqueous M. oleifera leaf and C. papaya seed extracts were associated with reductions in FEC, while the conventional dewormer demonstrated comparatively lower efficacy by Day 30. These findings are consistent with increasing reports of reduced benzimidazole effectiveness in small ruminant production systems in sub-Saharan Africa, where frequent use and limited drug rotation are common (Papadopoulos et al., 2021 ; Vineer et al., 2020 ). According to World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines, full anthelmintic efficacy requires a ≥ 95% reduction in FEC with a lower confidence limit above 90% (Levecke et al., 2022 ; Coles et al., 1992 ). In this present study, M. oleifera approached this threshold against Moniezia spp. but achieved only moderate reductions against H. contortus , while C. papaya exhibited a similar pattern with greater activity against cestodes than nematodes. These results indicate parasite-specific responses and suggest that the tested botanicals are unlikely to fully replace conventional anthelmintics but may contribute to integrated parasite control strategies (Torres-Acosta and Hoste, 2023 ; Eguale, 2021 ). The low FEC reductions observed for conventional dewormers are consistent with suspected reduced efficacy under field conditions characterized by repeated drug exposure, suboptimal dosing, and limited resistance surveillance (Kaminsky et al., 2022 ; Wondimu and Bayu, 2022 ). Although formal resistance confirmation was not undertaken, the magnitude of reduction observed aligns with earlier reports of compromised albendazole performance in small ruminants in sub-Saharan Africa (Eguale et al., 2009 ; Afful et al., 2010 ). Both plant extracts demonstrated progressive reductions in egg counts beyond Day 14 post-treatment. This delayed response contrasts with the rapid parasite clearance typically observed following administration of synthetic anthelmintics and may reflect cumulative or indirect modes of action, such as reduced worm fecundity or gradual disruption of parasite physiology (Belga et al., 2024 ; Hoste et al., 2006 ). Consequently, evaluation windows designed for synthetic compounds may underestimate the biological activity of botanical treatments, although confirmation in larger and more controlled studies is required (Levecke et al., 2022 ). The observed antiparasitic effects may be attributed to phytochemical constituents previously reported in M. oleifera leaves, including condensed tannins, flavonoids, alkaloids, and saponins, and in C. papaya seeds, such as benzyl isothiocyanate and proteolytic enzymes (Elghandour et al., 2023 ; Jato et al., 2022 ; Páez-León et al., 2022 ). These compounds have been associated with impaired larval development, altered metabolism, and reduced egg viability in gastrointestinal helminths. However, the absence of extract standardization and phytochemical profiling in the present study limits mechanistic interpretation and reproducibility across ecological settings. Sheep treated with M. oleifera showed the greatest numerical increase in body weight, although differences among treatment groups were not statistically significant. This trend may reflect partial parasite control combined with the nutritional properties of M. oleifera leaves, which are known to be rich in protein, minerals, and antioxidants (Pareek et al., 2023 ). Similar improvements in animal performance have been reported in studies involving tannin-containing forages and nutraceutical plant supplements in small ruminants (Bessell et al., 2018 ; Hoste et al., 2006 ). Several limitations warrant consideration. The modest sample size limited statistical power, particularly for detecting treatment × time interactions, while the single-site, single-season design restricts broader extrapolation. In addition, extract preparation was not chemically standardized, and long-term safety was not assessed. Despite these constraints, the study provides field-based evidence that locally available botanical resources may have a role in parasite management where conventional anthelmintic efficacy is reduced. Future research should prioritize extract standardization, dose optimisation, multi-site evaluation, and economic feasibility to better define the practical applicability of plant-based anthelmintics in small ruminant production systems (Eguale, 2021 ; Torres-Acosta and Hoste, 2023 ). CONCLUSION This study evaluated the anthelmintic activity of aqueous C. papaya seed and M. oleifera leaf extracts in sheep naturally infected with gastrointestinal helminths under field conditions in Ghana. Both plant extracts were associated with reductions in faecal egg counts of H. contortus and Moniezia spp., with M. oleifera showing higher and more consistent reductions by Day 30. The conventional dewormer exhibited low FECRT%, consistent with reduced efficacy under the prevailing management conditions. Although statistical significance across treatment groups was limited, the observed biological trends suggest that these botanicals may offer complementary value within integrated parasite management programmes, particularly in settings where conventional anthelmintic efficacy is compromised. Given the small sample size, single-site design, and lack of extract standardization, the findings should be interpreted cautiously. Further research involving larger populations, standardized preparations, and extended monitoring is required before recommendations for routine field use can be made. Declarations Ethical Approval Permission was sought from the Veterinary Service Directorate. All procedures were approved by the AAMUSTED Institutional Ethics Review Committee (Approval No. AAMUSTED/IERC/2024/031) and conducted according to the Three R’s principles (Replacement, Reduction, Refinement). Acknowledgements The authors express their sincere gratitude to the Regional Veterinary Officer and the Disease Investigation Farm/Regional Veterinary Laboratory (DIF/RVL) staff at Techiman for providing vital research and laboratory facilities. Special appreciation goes to veterinary professionals in Techiman, whose participation in the study provided valuable insights into the practical challenges of anthelmintic resistance. We also acknowledge the Akenten Appiah-Menka University of Skills Training and Entrepreneurial Development (AAMUSTED) Ethics Committee for their ethical oversight of the study. Consent for publication: Not applicable. Data Availability and Accessibility: The dataset supporting the findings of this study is publicly available and can be accessed at the following repository (Dwaah et. al 2025). Funding Statement: This research was conducted without any external funding. The authors declare that no financial support influenced the study outcomes. Conflict of Interest Statement: The authors declare no conflicts of interest. AUTHORS' CONTRIBUTIONS P.K.D.: conceptualized the study, coordinated fieldwork and laboratory analyses, collected data, and drafted the manuscript. N.Y.A.-B.: contributed to study design, ethical approval procedures, and critical revision of the manuscript. I.B. & D.D.Y. : contributed to interpretation of results and manuscript revision. H.D.-F. : contributed to study design and manuscript preparation. S.A.S. & H.A.L. : performed statistical analysis, contributed to data interpretation, and revised the manuscript. 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Vineer, H.R., Morgan, E.R., Hertzberg, H., Bartley, D.J., Bosco, A., Charlier, J., Chartier, C., Claerebout, E., de Waal, T., Hendrickx, G., Hinney, B., Höglund, J., Ježek, J., Kašný, M., Keane, O.M., Martínez-Valladares, M., Mateus, T.L., McIntyre, J., Mickiewicz, M., Muñoz, A.M., Phythian, C.J., Ploeger, H.W., Rataj, A.V., Skuce, P.J., Simin, S., Sotiraki, S., Spinu, M., Stuen, S., Thamsborg, S.M., Vadlejch, J., Varady, M., von Samson-Himmelstjerna, G., Rinaldi, L. (2020). Increasing importance of anthelmintic resistance in European livestock. Parasite 27, 69. Wondimu, A., Bayu, Y. (2022). Anthelmintic drug resistance of gastrointestinal nematodes in goats. Journal of Parasitology Research 2022, 4025902. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. 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2","display":"","copyAsset":false,"role":"figure","size":70382,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-8403819/v1/265dab28d61b5e494a72d718.png"},{"id":100572453,"identity":"9c6915ab-ef59-4b1e-b7ca-a85cd0660353","added_by":"auto","created_at":"2026-01-19 09:54:38","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":67840,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-8403819/v1/afefff671cb2846f9aa8e67e.png"},{"id":100595944,"identity":"e698bd43-846a-4c9a-81d7-59ae553fd87d","added_by":"auto","created_at":"2026-01-19 13:49:48","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":66046,"visible":true,"origin":"","legend":"\u003cp\u003eSee image above for figure legend\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-8403819/v1/75dd9afbd614ac76ba3dbee6.png"},{"id":104402664,"identity":"4236ac29-2e0c-4106-ab0d-0c11fab9102d","added_by":"auto","created_at":"2026-03-11 12:16:02","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1332213,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-8403819/v1/8a0518f7-d105-4b20-895b-a2091f1d3472.pdf"}],"financialInterests":"","formattedTitle":"Botanical Anthelmintics for Sustainable Sheep Production: In Vivo Efficacy of Carica papaya Seeds and Moringa oleifera Leaves Against Haemonchus contortus and Moniezia spp. in Ghana","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eSheep production constitutes an important component of smallholder livestock systems across Africa, contributing to household income, food security, and livelihood resilience. In West Africa, sheep are commonly managed under extensive and semi-intensive systems, where productivity is closely linked to environmental conditions and animal health management practices (Food and Agriculture Organization {FAO}, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Thornton \u0026amp; Herrero, 2021). Despite their adaptability, productivity in these systems is frequently constrained by parasitic diseases, with gastrointestinal helminths remaining among the most persistent challenges to sustainable sheep production.\u003c/p\u003e \u003cp\u003eAmong the helminths affecting small ruminants, \u003cem\u003eHaemonchus contortus\u003c/em\u003e (\u003cem\u003eH. contortus\u003c/em\u003e) and \u003cem\u003eMoniezia\u003c/em\u003e spp. are widely distributed in tropical and subtropical regions. \u003cem\u003eH. contortus\u003c/em\u003e is regarded as one of the most pathogenic nematodes of sheep due to its hematophagous nature, high fecundity, and capacity for rapid population expansion under favourable climatic conditions, leading to anaemia, reduced growth, and increased mortality (Kaminsky et al. \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Papadopoulos et al. \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Although infections with \u003cem\u003eMoniezia\u003c/em\u003e spp. are often considered of lower clinical importance, heavy or persistent burdens, particularly in young animals, may compromise feed efficiency and growth performance (Torres-Acosta \u0026amp; Hoste, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eControl of gastrointestinal helminths has historically depended on the regular use of broad-spectrum anthelmintic drugs. While this approach initially resulted in substantial improvements in livestock productivity, increasing reports of reduced drug efficacy have emerged globally (Vineer et al. \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Anthelmintic resistance (AR) is now widely recognized as a major constraint to parasite control in small ruminants, particularly in low- and middle-income countries where treatment is often frequent, diagnostics are limited, and dosing practices may be imprecise (Levecke et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Coles et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Under such conditions, reliance on a narrow range of conventional anthelmintics has raised concerns about the long-term sustainability of conventional parasite control programmes.\u003c/p\u003e \u003cp\u003eIn response to this challenge, attention has increasingly turned toward integrated parasite management approaches that combine strategic drug use with complementary control options. Ethnoveterinary medicine, which encompasses the use of locally available plant-based remedies, has gained renewed interest as a potential component of such strategies (Eguale \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Numerous medicinal plants have been reported to possess antiparasitic properties; however, their efficacy varies widely depending on plant species, preparation method, parasite species, and evaluation protocol (Belga et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Jato et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Moreover, many studies rely on in vitro assays or solvent-based extracts, limiting their applicability under field conditions typical of smallholder systems.\u003c/p\u003e \u003cp\u003e \u003cem\u003eMoringa oleifera\u003c/em\u003e (\u003cem\u003eM. oleifera\u003c/em\u003e) and \u003cem\u003eCarica papaya\u003c/em\u003e (\u003cem\u003eC. papaya\u003c/em\u003e) are widely distributed in Sub-Saharan regions and commonly used for nutritional and medicinal purposes. Experimental studies suggest that extracts from \u003cem\u003eM. oleifera\u003c/em\u003e leaves and \u003cem\u003eC. papaya\u003c/em\u003e seeds contain bioactive compounds capable of disrupting helminth development and survival (P\u0026aacute;ez-Le\u0026oacute;n et al. \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Cabral et al. \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). Nevertheless, in vivo evidence derived from naturally infected sheep, particularly using aqueous preparations representative of practical on-farm use in West Africa, remains limited.\u003c/p\u003e \u003cp\u003eThe present study therefore, assessed the anthelmintic activity of aqueous extracts of \u003cem\u003eM. oleifera\u003c/em\u003e leaves and \u003cem\u003eC. papaya\u003c/em\u003e seeds in sheep naturally infected with \u003cem\u003eH. contortus\u003c/em\u003e and \u003cem\u003eMoniezia\u003c/em\u003e spp. under field conditions in Ghana. By comparing their effects with a commonly used synthetic anthelmintic and an untreated control, the study seeks to generate applied evidence relevant to sustainable parasite control in tropical small ruminant production systems.\u003c/p\u003e"},{"header":"METHODOLOGY","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStudy Area\u003c/h2\u003e \u003cp\u003eThe study was conducted at the Disease Investigation Farm (DIF) in Techiman, Bono East Region, Ghana, an area known for its high prevalence of gastrointestinal parasitic infections in small ruminants, to investigate conventional dewormers by the Veterinary Directorate Service (VSD) of Ghana (VSD, 2019; MoFA, 2014). The site lies within the transitional ecological zone (7.59529\u0026deg; N, -1.93331\u0026deg; W), characterized by Guinea savannah and semi-deciduous vegetation. Techiman experiences a tropical savannah climate with annual rainfall of 1,200-1,500 mm and temperatures ranging from 22\u0026deg;C to 34\u0026deg;C during the study period (September-October 2024).\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003ePlant Materials and Preparation of Extracts\u003c/h3\u003e\n\u003cp\u003ePapaya seeds and moringa leaves were selected based on prior ethnoveterinary evidence and unpublished work by the research team. Mature papaya fruits and fresh moringa leaves were harvested from organically maintained plants at DIF.\u003c/p\u003e \u003cp\u003ePlant materials were shade-dried at ambient temperature (25\u0026ndash;30\u0026deg;C) for 10\u0026ndash;14 days, ground, and sieved (1 mm mesh). Fifty grams of each powdered sample were macerated in 500 mL of distilled water (1:10 w/v) for 12 hours, filtered through sterile muslin, and stored at 4\u0026deg;C for not more than 72 hours, following established protocols (Belga et al. \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Kedir, \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Jato et al. \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Castangna et al. 2021; Bessell et. al. 2021).\u003c/p\u003e\n\u003ch3\u003eDetermination of Safe Dosage\u003c/h3\u003e\n\u003cp\u003eA pilot dose-ranging trial was performed using 1\u0026ndash;10 mL/kg BW of each extract, observing for toxicity or behavioural changes over 72 hours. No adverse effects were recorded; therefore, 10 mL/kg BW was selected as the therapeutic dose for the main trial.\u003c/p\u003e\n\u003ch3\u003eExperimental Design\u003c/h3\u003e\n\u003cp\u003eA randomized controlled design was used involving 33 naturally infected sheep. Sample size determination followed the statistical framework of (Levecke et al. \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Coles et al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Animals were randomly allocated into four treatment groups as shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eExperimental groups, treatments, and administered dosages in naturally infected sheep.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"3\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eTreatments\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDosage\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eDistilled water (negative control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10 mL/kg BW\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003ePapaya seed extract\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10 mL/kg BW\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eMoringa leaf extract\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10 mL/kg BW\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAlbendazole (positive control)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10 mg/kg BW\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"3\"\u003e\u003cb\u003eFootnote\u003c/b\u003e:\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003col\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eBW\u0026thinsp;=\u0026thinsp;body weight.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003cspan\u003e \u003cli\u003e \u003cp\u003eAll treatments were administered orally on Day 0. Faecal samples were collected on Days 0, 5, 10, and 30 for FEC analysis.\u003c/p\u003e \u003c/li\u003e \u003c/span\u003e \u003c/ol\u003e \u003c/p\u003e\n\u003ch3\u003eFaecal Collection and Parasitological Examination\u003c/h3\u003e\n\u003cp\u003eFaecal samples were obtained directly from the rectum using standard operating procedures (Denwood et. al \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Quantitative FECs were performed using the McMaster technique (Mohammedsalih et. al, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e2025\u003c/span\u003e; Charles Sturt University, \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Playford \u0026amp; Besier, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Coles et. al, \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). Three grams of faeces were homogenized in 60 mL of saturated NaCl solution, sieved, and examined microscopically (\u0026times;10 objective) for helminth eggs within the grid area. Laboratory personnel were blinded to treatment allocation to minimize bias.\u003c/p\u003e \u003cp\u003eThe Faecal Egg Count Reduction Test (FECRT) was adopted from (Denwood et. al 2024; Coles et. al. \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1992\u003c/span\u003e): FECRT (%) = \u003cspan class=\"InlineEquation\"\u003e\u003cspan class=\"mathinline\"\u003e\\(\\:\\frac{\\left(Mean\\:FEC\\:before\\:treatment\\right)-\\left(Mean\\:FEC\\:after\\:treatment\\right)}{\\left(Mean\\:FEC\\:before\\:treatment\\right)}\\)\u003c/span\u003e\u003c/span\u003e \u0026times;100, using pre-treatment (Day 0) and post-treatment (Day 30) values for \u003cem\u003eHaemonchus contortus\u003c/em\u003e and \u003cem\u003eMoniezia spp.\u003c/em\u003e\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eStatistical Analysis\u003c/h2\u003e \u003cp\u003eDescriptive statistics (mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) were computed using Microsoft Excel 2019. Data normality was assessed with the Shapiro-Wilk test. One-way ANOVA was used to compare mean FECs among groups, with Tukey\u0026rsquo;s Honest Significant Difference (HSD) post hoc test for pairwise comparisons (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). Two-way ANOVA evaluated treatment \u0026times; time interaction effects, and paired t-tests compared pre- and post-treatment values within groups. Post hoc power analysis (α\u0026thinsp;=\u0026thinsp;0.05) confirmed sample adequacy (\u0026gt;\u0026thinsp;0.8).\u003c/p\u003e \u003c/div\u003e"},{"header":"RESULTS","content":"\u003cdiv id=\"Sec10\" class=\"Section2\"\u003e \u003ch2\u003eHelminth Load before Treatment\u003c/h2\u003e \u003cp\u003eAt baseline, 60.6% (20/33) of sheep were co-infected with \u003cem\u003eH. contortus\u003c/em\u003e and \u003cem\u003eMoniezia\u003c/em\u003e spp., while 30.3% (10/33) were infected with \u003cem\u003eH. contortus\u003c/em\u003e alone and 9.1% (3/33) with \u003cem\u003eMoniezia\u003c/em\u003e spp. alone (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). Parasite burdens ranged from moderate to high according to Veterinary Services Directorate classification criteria. Rams showed a higher mean FEC than females and younger animals, although these differences were not subjected to statistical testing due to sample size constraints. There were no significant differences in baseline FECs among treatment groups for either \u003cem\u003eH. contortus\u003c/em\u003e (p\u0026thinsp;=\u0026thinsp;0.72) or \u003cem\u003eMoniezia\u003c/em\u003e spp. (p\u0026thinsp;=\u0026thinsp;0.68), indicating comparability across groups before treatment.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePre-treatment distribution of \u003cem\u003eH. contortus\u003c/em\u003e and \u003cem\u003eMoniezia\u003c/em\u003e spp. infections by sex and age category (n\u0026thinsp;=\u0026thinsp;33 sheep).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSex and Age\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eNo. Sheep\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003eType of infection\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cb\u003eH. contortus\u003c/b\u003e\u003c/p\u003e \u003cp\u003e\u003cb\u003eonly\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cb\u003eMoniezia\u003c/b\u003e \u003cb\u003espp\u003c/b\u003e.\u003c/p\u003e \u003cp\u003e\u003cb\u003eonly\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cb\u003eBoth helminths\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRam\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eEwe\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYoung Ram\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYoung Ewe\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003en (%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e33 (100)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10 (30.3)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3 (9.1)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20 (60.6)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"5\"\u003e\u003cb\u003eFootnote\u003c/b\u003e: Both \u003cem\u003eM. expansa\u003c/em\u003e and \u003cem\u003eM. benedeni\u003c/em\u003e were detected and are reported collectively as \u003cem\u003eMoniezia\u003c/em\u003e spp.\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eFECRT of Aqueous\u003c/b\u003e \u003cb\u003eMoringa oleifera\u003c/b\u003e \u003cb\u003eLeaves and\u003c/b\u003e \u003cb\u003eCarica papaya\u003c/b\u003e \u003cb\u003eSeeds against\u003c/b\u003e \u003cb\u003eH. contortus\u003c/b\u003e\u003c/p\u003e \u003cp\u003eReductions in \u003cem\u003eH. contortus\u003c/em\u003e FEC differed among treatment groups over time (Fig.\u0026nbsp;1). From \u003cem\u003eFig.\u0026nbsp;2\u003c/em\u003e, by Day 30 post-treatment, sheep receiving aqueous \u003cem\u003eM. oleifera\u003c/em\u003e leaf extract showed a mean FECRT of 60.9% (95% CI: 52.2\u0026ndash;69.5), while those treated with aqueous \u003cem\u003eC. papaya\u003c/em\u003e seed extract achieved a reduction of 51.3% (95% CI: 43.1\u0026ndash;59.4). Conventional dewormer treatment resulted in a mean FECRT of 43.9% (95% CI: 35.4\u0026ndash;52.1). In contrast, mean FEC in the untreated control group increased over the same period.\u003c/p\u003e \u003cp\u003eOne-way ANOVA indicated significant differences in mean FECs between treated groups and the untreated control at Day 30 (p\u0026thinsp;\u0026lt;\u0026thinsp;0.05). However, when evaluated using two-way ANOVA, neither the main effects of treatment (p\u0026thinsp;=\u0026thinsp;0.219) nor time (p\u0026thinsp;=\u0026thinsp;0.117) nor their interaction (p\u0026thinsp;=\u0026thinsp;0.995) were statistically significant.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cb\u003eFECRT of aqueous\u003c/b\u003e \u003cb\u003eoleifera\u003c/b\u003e \u003cb\u003eleaves and aqueous C. papaya seeds against\u003c/b\u003e \u003cb\u003eMoniezia\u003c/b\u003e \u003cb\u003espp.\u003c/b\u003e\u003c/p\u003e \u003cp\u003eA progressive reduction in \u003cem\u003eMoniezia\u003c/em\u003e spp. FEC was observed in sheep treated with both plant extracts (Fig.\u0026nbsp;3). From \u003cem\u003eFig.\u0026nbsp;4\u003c/em\u003e, by Day 10, aqueous \u003cem\u003eM. oleifera\u003c/em\u003e leaf extract achieved a FECRT of 52.6% \u0026plusmn; 6.7 (95% CI: 40.2\u0026ndash;65.1), which increased to 92.1% \u0026plusmn; 4.8 (95% CI: 83.6\u0026ndash;100.0) by Day 30. Sheep treated with aqueous \u003cem\u003eC. papaya\u003c/em\u003e seed extract showed mean FECRTs of 35.3% \u0026plusmn; 5.5 (95% CI: 24.1\u0026ndash;46.3) at Day 10 and 82.4% \u0026plusmn; 6.2 (95% CI: 70.1\u0026ndash;94.6) at Day 30.\u003c/p\u003e \u003cp\u003eConventional dewormer treatment resulted in a mean FECRT of 50.0% \u0026plusmn; 7.3 (95% CI: 33.5\u0026ndash;66.5) by Day 30. In contrast, the untreated control group exhibited a mean increase in egg counts of 111.8% \u0026plusmn; 9.2% over the study period.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eEffect of Treatment Type and Time on Anthelmintic Efficacy\u003c/h2\u003e \u003cp\u003eTwo-way ANOVA was used to assess the effects of treatment, time, and their interaction on FECRT% (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). No statistically significant main effects of treatment or time were detected, and no considerable treatment \u0026times; time interaction was observed (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05 for all comparisons). These results indicate substantial within-group variability and suggest that treatment-associated changes over time were not consistent across all animals.\u003c/p\u003e \u003cp\u003eExploratory post hoc comparisons suggested within-group reductions in egg counts over time for sheep receiving \u003cem\u003eM. oleifera\u003c/em\u003e. Still, these findings should be interpreted cautiously, given the absence of significant overall interaction effects.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTwo-way ANOVA assessing the effects of treatment, time, and their interaction on faecal egg count reduction (%).\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"5\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSource of Variation\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSum of Squares\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003edf\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eF-Statistic\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003ep-Value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment Type\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2276.78\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.219\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTime\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3468.11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2.75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.117\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment \u0026times; Time (Interaction)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e115.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.046\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e0.995\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eResidual (Error)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5677.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cstrong\u003eInterpretation\u003c/strong\u003e \u003cp\u003e \u003cem\u003eLack of significant interaction (p\u0026thinsp;=\u0026thinsp;0.995) implies consistent treatment effect over time.\u003c/em\u003e \u003c/p\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003eWeight Gain Change\u003c/h2\u003e \u003cp\u003eMean body weight increased over time in all treated groups (Table\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e). Sheep treated with aqueous \u003cem\u003eM. oleifera\u003c/em\u003e leaf extract exhibited the greatest numerical increase in mean body weight over the 30 days (mean gain: 4.7 kg), followed by those treated with aqueous \u003cem\u003eC. papaya\u003c/em\u003e seed extract and Conventional dewormer. Sheep in the untreated control group showed minimal change in body weight over the same period. One-way ANOVA did not detect statistically significant differences in body weight among treatment groups at any sampling point (p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMean body weight (kg, mean\u0026thinsp;\u0026plusmn;\u0026thinsp;SD) of sheep by treatment group at each sampling time.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\"\u0026plusmn;\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c2\" namest=\"c1\"\u003e \u003cp\u003eTreatment\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c5\" namest=\"c3\"\u003e \u003cp\u003eDays\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eP-value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eConventional Dewormer\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e24.13\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e25.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e26.13\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e27.63\u0026thinsp;\u0026plusmn;\u0026thinsp;2.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.770\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAqueous moringa leaves\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e24.5\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e25.63\u0026thinsp;\u0026plusmn;\u0026thinsp;3.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e27.06\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e29.19\u0026thinsp;\u0026plusmn;\u0026thinsp;3.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.720\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAqueous papaya seeds\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e26\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e27.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e28.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e29.06\u0026thinsp;\u0026plusmn;\u0026thinsp;2.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.886\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eDistilled Water\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c2\"\u003e \u003cp\u003e26\u0026thinsp;\u0026plusmn;\u0026thinsp;2.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c3\"\u003e \u003cp\u003e25.44\u0026thinsp;\u0026plusmn;\u0026thinsp;2.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c4\"\u003e \u003cp\u003e26.22\u0026thinsp;\u0026plusmn;\u0026thinsp;2.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\"\u0026plusmn;\" colname=\"c5\"\u003e \u003cp\u003e26.39\u0026thinsp;\u0026plusmn;\u0026thinsp;2.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0.995\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003cb\u003eFootnote\u003c/b\u003e: No significant differences in body weight were detected among treatment groups (one-way ANOVA; p\u0026thinsp;\u0026gt;\u0026thinsp;0.05).\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003ePost Hoc Analysis\u003c/h2\u003e \u003cp\u003eAlthough the overall treatment \u0026times; time interaction was not significant, exploratory pairwise comparisons indicated within-group changes over time, as shown in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. These findings should be interpreted cautiously, given the absence of statistically significant main effects.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eTukey\u0026rsquo;s HSD pairwise comparisons of faecal egg count reduction (%) across sampling days.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGroup 1\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eGroup 2\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMean Diff\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003ep-Value\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e95% CI (LP)\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eSignificant\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D-Day 0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC.D-Day10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.939\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-69.36 to 129.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D-Day 0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eC.D-Day5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.998\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-82.36 to 116.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eC.D-Day10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eACPS-Day 10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e-20.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.994\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-119.36 to 79.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eACPS-Day 0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAMOL- Day 10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-41.86 to 156.86\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eACPS-Day 5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAMOL-Day 5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e21\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.992\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e-78.36 to 120.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003ctfoot\u003e \u003ctr\u003e\u003ctd colspan=\"6\"\u003e\u003cb\u003eFootnote\u003c/b\u003e: i. CD is Conventional Dewormer, ACPS is Aqueous \u003cem\u003eCarica Papaya\u003c/em\u003e seeds, and AMOL is Aqueous \u003cem\u003eMoringa oleifera\u003c/em\u003e leaves\u003c/td\u003e\u003c/tr\u003e \u003c/tfoot\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eii. Pairwise comparisons are exploratory and should be interpreted in the context of the non-significant overall ANOVA.\u003c/p\u003e \u003c/div\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eThis study assessed the in vivo efficacy of two ethnoveterinary plant extracts against gastrointestinal helminths in sheep under natural grazing conditions. Both aqueous \u003cem\u003eM. oleifera\u003c/em\u003e leaf and \u003cem\u003eC. papaya\u003c/em\u003e seed extracts were associated with reductions in FEC, while the conventional dewormer demonstrated comparatively lower efficacy by Day 30. These findings are consistent with increasing reports of reduced benzimidazole effectiveness in small ruminant production systems in sub-Saharan Africa, where frequent use and limited drug rotation are common (Papadopoulos et al., \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Vineer et al., \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e2020\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAccording to World Association for the Advancement of Veterinary Parasitology (WAAVP) guidelines, full anthelmintic efficacy requires a\u0026thinsp;\u0026ge;\u0026thinsp;95% reduction in FEC with a lower confidence limit above 90% (Levecke et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Coles et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). In this present study, \u003cem\u003eM. oleifera\u003c/em\u003e approached this threshold against \u003cem\u003eMoniezia\u003c/em\u003e spp. but achieved only moderate reductions against \u003cem\u003eH. contortus\u003c/em\u003e, while \u003cem\u003eC. papaya\u003c/em\u003e exhibited a similar pattern with greater activity against cestodes than nematodes. These results indicate parasite-specific responses and suggest that the tested botanicals are unlikely to fully replace conventional anthelmintics but may contribute to integrated parasite control strategies (Torres-Acosta and Hoste, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Eguale, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe low FEC reductions observed for conventional dewormers are consistent with suspected reduced efficacy under field conditions characterized by repeated drug exposure, suboptimal dosing, and limited resistance surveillance (Kaminsky et al., \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; Wondimu and Bayu, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Although formal resistance confirmation was not undertaken, the magnitude of reduction observed aligns with earlier reports of compromised albendazole performance in small ruminants in sub-Saharan Africa (Eguale et al., \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Afful et al., \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eBoth plant extracts demonstrated progressive reductions in egg counts beyond Day 14 post-treatment. This delayed response contrasts with the rapid parasite clearance typically observed following administration of synthetic anthelmintics and may reflect cumulative or indirect modes of action, such as reduced worm fecundity or gradual disruption of parasite physiology (Belga et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2024\u003c/span\u003e; Hoste et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Consequently, evaluation windows designed for synthetic compounds may underestimate the biological activity of botanical treatments, although confirmation in larger and more controlled studies is required (Levecke et al., \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe observed antiparasitic effects may be attributed to phytochemical constituents previously reported in \u003cem\u003eM. oleifera\u003c/em\u003e leaves, including condensed tannins, flavonoids, alkaloids, and saponins, and in \u003cem\u003eC. papaya\u003c/em\u003e seeds, such as benzyl isothiocyanate and proteolytic enzymes (Elghandour et al., \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e2023\u003c/span\u003e; Jato et al., \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e2022\u003c/span\u003e; P\u0026aacute;ez-Le\u0026oacute;n et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). These compounds have been associated with impaired larval development, altered metabolism, and reduced egg viability in gastrointestinal helminths. However, the absence of extract standardization and phytochemical profiling in the present study limits mechanistic interpretation and reproducibility across ecological settings.\u003c/p\u003e \u003cp\u003eSheep treated with \u003cem\u003eM. oleifera\u003c/em\u003e showed the greatest numerical increase in body weight, although differences among treatment groups were not statistically significant. This trend may reflect partial parasite control combined with the nutritional properties of \u003cem\u003eM. oleifera\u003c/em\u003e leaves, which are known to be rich in protein, minerals, and antioxidants (Pareek et al., \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2023\u003c/span\u003e). Similar improvements in animal performance have been reported in studies involving tannin-containing forages and nutraceutical plant supplements in small ruminants (Bessell et al., \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Hoste et al., \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e2006\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eSeveral limitations warrant consideration. The modest sample size limited statistical power, particularly for detecting treatment \u0026times; time interactions, while the single-site, single-season design restricts broader extrapolation. In addition, extract preparation was not chemically standardized, and long-term safety was not assessed. Despite these constraints, the study provides field-based evidence that locally available botanical resources may have a role in parasite management where conventional anthelmintic efficacy is reduced. Future research should prioritize extract standardization, dose optimisation, multi-site evaluation, and economic feasibility to better define the practical applicability of plant-based anthelmintics in small ruminant production systems (Eguale, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Torres-Acosta and Hoste, \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e2023\u003c/span\u003e).\u003c/p\u003e"},{"header":"CONCLUSION","content":"\u003cp\u003eThis study evaluated the anthelmintic activity of aqueous \u003cem\u003eC. papaya\u003c/em\u003e seed and \u003cem\u003eM. oleifera\u003c/em\u003e leaf extracts in sheep naturally infected with gastrointestinal helminths under field conditions in Ghana. Both plant extracts were associated with reductions in faecal egg counts of \u003cem\u003eH. contortus\u003c/em\u003e and \u003cem\u003eMoniezia\u003c/em\u003e spp., with \u003cem\u003eM. oleifera\u003c/em\u003e showing higher and more consistent reductions by Day 30. The conventional dewormer exhibited low FECRT%, consistent with reduced efficacy under the prevailing management conditions.\u003c/p\u003e \u003cp\u003eAlthough statistical significance across treatment groups was limited, the observed biological trends suggest that these botanicals may offer complementary value within integrated parasite management programmes, particularly in settings where conventional anthelmintic efficacy is compromised. Given the small sample size, single-site design, and lack of extract standardization, the findings should be interpreted cautiously. Further research involving larger populations, standardized preparations, and extended monitoring is required before recommendations for routine field use can be made.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eEthical Approval\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003ePermission was sought from the Veterinary Service Directorate. All procedures were approved by the AAMUSTED Institutional Ethics Review Committee (Approval No. AAMUSTED/IERC/2024/031) and conducted according to the Three R’s principles (Replacement, Reduction, Refinement).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors express their sincere gratitude to the Regional Veterinary Officer and the Disease Investigation Farm/Regional Veterinary Laboratory (DIF/RVL) staff at Techiman for providing vital research and laboratory facilities. Special appreciation goes to veterinary professionals in Techiman, whose participation in the study provided valuable insights into the practical challenges of anthelmintic resistance. We also acknowledge the Akenten Appiah-Menka University of Skills Training and Entrepreneurial Development (AAMUSTED) Ethics Committee for their ethical oversight of the study.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConsent for publication:\u0026nbsp;\u003c/strong\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability and Accessibility:\u0026nbsp;\u003c/strong\u003eThe dataset supporting the findings of this study is publicly available and can be accessed at the following repository (Dwaah et. al 2025).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eFunding Statement:\u0026nbsp;\u003c/strong\u003eThis research was conducted without any external funding. The authors declare that no financial support influenced the study outcomes.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConflict of Interest Statement:\u0026nbsp;\u003c/strong\u003eThe authors declare no conflicts of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAUTHORS' CONTRIBUTIONS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eP.K.D.:\u0026nbsp;\u003c/strong\u003econceptualized the study, coordinated fieldwork and laboratory analyses, collected data, and drafted the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eN.Y.A.-B.:\u003c/strong\u003e contributed to study design, ethical approval procedures, and critical revision of the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eI.B. \u0026amp; D.D.Y.\u003c/strong\u003e: contributed to interpretation of results and manuscript revision.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eH.D.-F.\u003c/strong\u003e: contributed to study design and manuscript preparation.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eS.A.S. \u0026amp; H.A.L.\u003c/strong\u003e: performed statistical analysis, contributed to data interpretation, and revised the manuscript.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eAll authors reviewed and approved the final version of the manuscript.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\u003cli\u003e\u003cspan\u003eAfful, E.D.D., Turkson, J.D., Biney, A.K. 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Accra, Ghana: Ministry of Food and Agriculture.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eVineer, H.R., Morgan, E.R., Hertzberg, H., Bartley, D.J., Bosco, A., Charlier, J., Chartier, C., Claerebout, E., de Waal, T., Hendrickx, G., Hinney, B., H\u0026ouml;glund, J., Ježek, J., Kašn\u0026yacute;, M., Keane, O.M., Mart\u0026iacute;nez-Valladares, M., Mateus, T.L., McIntyre, J., Mickiewicz, M., Mu\u0026ntilde;oz, A.M., Phythian, C.J., Ploeger, H.W., Rataj, A.V., Skuce, P.J., Simin, S., Sotiraki, S., Spinu, M., Stuen, S., Thamsborg, S.M., Vadlejch, J., Varady, M., von Samson-Himmelstjerna, G., Rinaldi, L. (2020). Increasing importance of anthelmintic resistance in European livestock. \u003cem\u003eParasite\u003c/em\u003e 27, 69.\u003c/span\u003e\u003c/li\u003e \u003cli\u003e\u003cspan\u003eWondimu, A., Bayu, Y. (2022). Anthelmintic drug resistance of gastrointestinal nematodes in goats. \u003cem\u003eJournal of Parasitology Research\u003c/em\u003e 2022, 4025902.\u003c/span\u003e\u003c/li\u003e\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Sheep, Helminths, Anthelmintic resistance, Ethnoveterinary Medicine, Faecal Egg Count Reduction, Anthelmintic Efficacy","lastPublishedDoi":"10.21203/rs.3.rs-8403819/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-8403819/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eGastrointestinal helminths, particularly \u003cem\u003eHaemonchus contortus (H. contortus)\u003c/em\u003e and \u003cem\u003eMoniezia\u003c/em\u003e spp., continue to constrain sheep productivity in sub-Saharan Africa. Increasing reports of reduced efficacy of conventional anthelmintics underline the need to evaluate sustainable alternative control options. This study assessed the in vivo anthelmintic efficacy of aqueous extracts of \u003cem\u003eCarica papaya\u003c/em\u003e seeds (CPS) and \u003cem\u003eMoringa oleifera\u003c/em\u003e leaves (MOL) in naturally infected sheep under field conditions in Ghana. Thirty-three sheep were allocated to four treatment groups: CPS, MOL, conventional dewormer, and distilled water. Treatments were administered orally on Day 0, and faecal egg counts (FEC) were determined on Days 0, 5, 10, and 30 using the McMaster technique. Anthelmintic efficacy was evaluated using the faecal egg count reduction test (FECRT), and body weight changes were monitored over the study period. By Day 30, MOL achieved a mean FECRT of 92.1% against \u003cem\u003eMoniezia\u003c/em\u003e spp. and 60.9% against \u003cem\u003eH. contortus\u003c/em\u003e, while CPS achieved reductions of 82.4% and 51.3%, respectively. The conventional dewormer showed lower efficacy (\u0026le;\u0026thinsp;50%) for both parasites. Sheep receiving MOL exhibited the greatest mean body weight gain (4.7 kg), although differences among treatments were not statistically significant. Egg counts increased markedly in the distilled water group. The findings indicate that MOL and CPS exhibit moderate to high anthelmintic activity under field conditions and may offer complementary options for helminth control in sheep production systems where resistance to conventional anthelmintics is of concern, which may contribute to integrated parasite management. Further studies with larger sample sizes and standardized extract characterization are warranted.\u003c/p\u003e","manuscriptTitle":"Botanical Anthelmintics for Sustainable Sheep Production: In Vivo Efficacy of Carica papaya Seeds and Moringa oleifera Leaves Against Haemonchus contortus and Moniezia spp. in Ghana","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2026-01-19 09:54:34","doi":"10.21203/rs.3.rs-8403819/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"84970b66-3dd5-4ca8-9e59-22f81cfc0647","owner":[],"postedDate":"January 19th, 2026","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2026-03-05T17:05:58+00:00","versionOfRecord":[],"versionCreatedAt":"2026-01-19 09:54:34","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-8403819","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-8403819","identity":"rs-8403819","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}