The effects of age, mature oocyte number, and cycle number on cumulative live birth rates after planned oocyte cryopreservation.

PURPOSE: To examine the effects of age, mature oocyte number, and cycle number on cumulative live birth rates after planned oocyte cryopreservation (OC), with the goal of developing a patient counselling tool. METHODS: We performed a retrospective cohort study of all patients with ≥ 1 autologous oocyte thaw at our university-affiliated fertility center before 12/31/2023. Patients were included if they (1) had a live birth or ongoing pregnancy > 12 weeks from OC, or (2) used all oocytes and euploid/untested embryos from OC. Primary outcome was cumulative live birth / ongoing pregnancy rate (CLBR). RESULTS: 527 patients with 1 OC cycle, 149 patients with 2 OC cycles, and 55 patients with ≥ 3 OC cycles were included. Overall CLBR was 43%. CLBR was > 70% among patients who thawed ≥ 20 mature oocytes that were cryopreserved at age < 38 years. Multiple logistic regression showed that age at first OC and total number of mature oocytes thawed independently predicted CLBR, but number of OC cycles did not. CONCLUSION: Patients must be counselled that younger age at OC and more mature oocytes improve CLBR. However, additional OC cycles do not independently improve CLBR. Our results can help patients decide whether to pursue additional OC cycles to obtain more oocytes.

Utilization of green lentil wastewater as egg replacer in green lentil flour based muffins.

Successful pretreatments for green lentil wastewater (GLW) were developed to substitute egg. Water to lentil ratio and microwave pretreatment were found to affect foam and emulsion quality, while the addition of salt had no effect on foam and emulsion quality of GLW. The GLW obtained at optimum preconditions was used in the determination of best formulation for muffin quality. Oven type, green lentil flour ratio, GLW ratio leading to the maximum moisture content, volume index, total phenolic content, percent area of air cells, and minimum ΔE values with a constraint of control muffin's hardness were determined. Conventional oven baking with the formulation of 5.71% green lentil flour and 18.15% GLW produced comparable product with wheat flour and egg formulation. This study proved that discarded GLW can be used as a substitute for egg, which is an expensive ingredient in bakery.

Patterns of gastrointestinal parasite infections in bighorn sheep, Ovis canadensis, with respect to host sex and seasonality.

Males and females in sexually dimorphic species show differences in their physiology and behaviour due to differences in energetic investment into reproduction and soma. This means that the two sexes may show different patterns of parasitism at different times of the year. In this study, we evaluate the abundance of fecal eggs and larvae of 5 parasite types (Strongyles, Nematodirus spp., Marshallagia marshalli., Protostrongylus spp. lungworms, and Eimeria spp.) in relation to season and sex in Rocky Mountain bighorn sheep (Ovis canadensis). We use fecal egg counts (FEC) as a proxy for infection intensity. Parasite FECs differed between male and female bighorn sheep and varied with season. We found pronounced fluctuations in fecal egg counts of various parasite species in males and females across different seasons and reproductive stages. Strongyle counts were significantly higher during late gestation and lactation/summer, and particularly pronounced in males. Nematodirus counts were highest during late gestation in females and during the rut in males. Marshallagia counts peaked during late gestation in females and during the rut in males. Protostrongylus spp. lungworm counts were highest during late gestation in females and in males during lactation/summer and the rut. Eimeria oocyst counts varied across seasons, with higher counts in males during the rut and in females during winter and late gestation. Additionally, significant differences in Strongyle counts were observed between coursing and tending rams, with tending rams exhibiting higher counts. We discuss why the sexes might differ in FECs and suggest that differences between FECs of the parasites across seasons may be due to different life cycles and cold tolerance of the parasites themselves.

Utilizing nutrient type compounds as anti-bacterial compounds: arginine and cysteine inhibit Salmonella survival in egg white.

Because of growing levels of antibiotic resistance, new methods to combat bacteria are needed. We hypothesized that because bacteria evolved to survive in specific environments, the addition of compounds, including nutrient type compounds, to an environment, might result in a modification of that environment that will disrupt bacterial growth or in maladaptive bacterial behavior, i.e., gene expression. As a proof of concept, we focused on the egg white environment and the pathogen Salmonella. Despite egg white's antibacterial nature, Salmonella is able to survive and grow in egg white, and this ability of Salmonella leads to infection of chicks and humans. Here, the 20 L-amino-acids were screened for their ability to affect the growth of Salmonella in egg white. L-arginine and L-cysteine were found to reduce growth in egg white in physiologically relevant concentrations. To determine the mechanism behind L-arginine inhibition TnSeq was utilized. TnSeq identified many Salmonella genes required for survival in egg white including genes required for iron import, biotin synthesis, stress responses, cell integrity, and DNA repair. However, a comparison of Salmonella in egg white with and without L-arginine identified only a few differences in the frequency of transposon insertions, including the possible contribution of perturbations in the cell envelope to the inhibition mechanism. Finally, both D-arginine and D-cysteine were found to inhibit Salmonella in egg white. This implied that the effect of arginine and cysteine in egg white is chemical rather than biological, likely on the egg white environment or on the bacterial outer membrane. To conclude, these results show that this approach of addition of compounds, including nutrient type compounds, to an environment can be used to limit bacterial growth. Importantly, these compounds have no inherent anti-bacterial properties, are used as nutrients by animals and bacteria, and only become anti-bacterial in a specific environmental context. Future research screening for the effects of compounds in relevant environments might uncover new ways to reduce pathogen levels in the poultry industry and beyond.

Live birth after vitrification of oocytes from capacitation in vitro maturation.

Female fertility preservation is a rapidly growing field in medicine. Oocyte cryopreservation and assisted reproductive technique with vitrified-warmed oocytes have been successful with in vivo matured oocytes after conventional ovarian stimulation protocols. The use of in vitro matured oocytes after vitrification and warming has been limited. Capacitation in vitro maturation (CAPA-IVM) represents the latest refinement of IVM protocols and provides in vitro matured oocytes with improved competence. This case report describes the first successful live birth following oocyte vitrification from a CAPA-IVM cycle. This milestone achievement holds a significant promise to expand fertility preservation options and improve accessibility for women wishing to cryopreserve their eggs for future use.

Effects of low dietary calcium and lipopolysaccharide challenges on production performance, eggshell quality, and bone metabolism of laying hens.

Dietary calcium supply is essential for bone development and egg production in laying hens. This study investigated the effects of low dietary calcium and lipopolysaccharide (LPS) induced immune challenge in aged laying hens. A total of thirty-two Hy-Line Brown laying hens at 80 weeks old with an average laying rate of 62% were randomly divided into two groups and fed a normal calcium diet (3.57% Ca, NCA) or low calcium diet (2.08% Ca, LCA). At 88 weeks, the experiment was designed using a 2 × 2 factorial arrangement, and hens were intraperitoneally injected with saline (SAL) or LPS (0.5 mg/kg, 0.5 mg/kg, or 1.5 mg/kg body weight) once every 48 h intervals over 5 days. Production performance, egg quality, and bone physiology were evaluated. Results showed that LPS challenge decreased the hen-day egg production, egg mass, and eggshell traits (p < 0.05), but increased (p < 0.05) the calcium content of the tibia compared to SAL-injected hens. LCA diet decreased (p < 0.05) the hen-day egg production, and eggshell traits such as weight, percentage, strength, and thickness compared to the NCA diet. LCA diet increased the serum alkaline phosphatase (ALP) activity (p < 0.01) and tibial expression of ALP (p < 0.05) compared to NCA diet. LPS injection suppressed both the serum ALP activity (p < 0.05) and tibial expression of ALP (p < 0.001) compared to SAL injection. Furthermore, LPS injection increased (p < 0.05) the expression of both pro and anti-inflammatory cytokines in the spleen and tibia. The expression of cathepsin K ( Cts K ) and matrix metalloproteinase 9 ( MMP-9 ) were downregulated by LPS injection (p < 0.001). Broken and shell-less egg production and calcium content of eggshell, as well as tibial mRNA expression of osteocalcin ( Ocn ), tumor necrosis factor-alpha ( TNF-α ) and tartrate-resistant acid phosphatase ( TRAP ) were affected by the interaction (p < 0.05) of diet and injection. Therefore, this study demonstrated that to certain extents, low dietary calcium and LPS challenge dysregulated bone homeostasis and metabolism, with detrimental effects on the performance and eggshell quality of aged laying hens.

Increasing the pasteurization temperature of liquid egg white by phosphorylation modification: Exploring the mechanism and application.

Pasteurization is an effective sterilization technique for the treatment of liquid egg white (LEW), but the pasteurization temperature is generally limited because increased temperature can lead to aggregation of the proteins and affect their processing properties. In this study, phosphorylation modification was used to increase the thermal stability and pasteurization temperature of LEW, aiming to enhance the pasteurization sterilizing effect. The FT-IR results showed that the phosphate groups were successfully grafted into protein molecules, improving the order degree of protein molecules. In this case, the pasteurization temperature of LEW increased from 58 °C to 61 °C, without accompanying thermal aggregation. The molecular structural results suggested that the enhanced thermal stability was attributed to the decreased average particle size and the increased electrostatic repulsion between protein molecules, which largely reduced the turbidity of LEW during pasteurization treatment. Meanwhile, this process was dominated by noncovalent interactions (hydrophobic interactions and hydrogen bonding). Furthermore, the phosphorylation modification can synchronously improve emulsifying and foaming properties of LEW. Therefore, this work suggested that phosphorylation has great potential to improve thermal stability and pasteurization temperature of LEW, which can be utilized to extend its sterilizing effect and shelf life.

Translucent eggs of laying hens: a review.

Eggshell quality is a significant characteristic that influences consumer preferences. Eggshell translucency is a common defect in the appearance of eggshells, which are characterized by gray spots that are visible to the naked eye under natural light. The presence of various defects resulting from eggshell translucency has caused a decrease in consumer willingness to purchase eggs, leading to considerable economic losses in the egg industry. Although the impact of eggshell translucency on food safety, egg quality, and hatchability has been extensively investigated, the classification and causes of eggshell translucency remain unclear and lack a systematic summary. In recent studies, new interpretations of evaluation methods and causes of eggshell translucency have been proposed, along with numerous innovative solutions. Therefore, this paper aims to provide a comprehensive review of the evaluation methods, classification, causes, effects, and influencing factors of eggshell translucency and to summarize the treatments for translucent eggs. We believe that this review will serve as a valuable reference for researchers involved in the study of translucent eggs.

Phylogenetic variations in a novel family of hyperstable apple snail egg proteins: insights into structural stability and functional trends.

The relationship between protein stability and functional evolution is little explored in proteins purified from natural sources. Here we investigate a novel family of egg proteins (Perivitellin-1, PV1) from Pomacea snails. Their remarkable stability and clade-related functions in most derived clades (Canaliculata and Bridgesii) make them excellent candidates for exploring this issue. To that aim, we studied PV1 (PpaPV1) from the most basal lineage, Flagellata. PpaPV1 displays unparalleled structural and kinetic stability, surpassing PV1s from derived clades, ranking among the most hyperstable proteins documented in nature. Its spectral features contribute to a pale egg coloration, exhibiting a milder glycan binding lectin activity with a narrower specificity than PV1s from the closely related Bridgesii clade. These findings provide evidence for substantial structural and functional changes throughout the genus' PV1 evolution. We observed that structural and kinetic stability decreases in a clade-related fashion and was associated with large variations in defensive traits. For instance, pale PpaPV1 lectin turns potent in the Bridgesii clade adversely affecting gut morphology, while giving rise to brightly colored PV1s providing eggs with a conspicuous, likely warning signal in the Canaliculata clade. This work provides a comprehensive comparative analysis of PV1s from various apple snail species within a phylogenetic framework, offering insights into the interplay among their structural features, stability profiles, and functional roles. More broadly, our work provides one of the first examples from natural evolution showing the crucial link among protein structure, stability, and evolution of new functions.

Egg protein exposure estimation in risk assessment for Japanese food allergy labeling.

To assess the risk of food allergies in foods processed under the Japanese food labeling system, estimating exposure to hidden allergens is necessary. We assessed exposure to egg protein in foods processed according to the Japanese food labeling system. First, we estimated the concentration distribution of egg protein by Bayesian methods using data from the literature and the measurement of food products with precautional declarations in the labeling margin. We then estimated the food-intake portion-size distribution under two scenarios: soft drink consumption as an example of single, high-intake consumption, and confections, which are frequently consumed by children, as a realistic example of low-intake consumption. Finally, we estimated the distribution of unexpected intake of egg proteins in the form of single consumption. The mean exposure to egg protein under the high-intake scenario was estimated to be 0.0164 mg for 1-15-year-olds, 0.0171 mg for 4-15-year-olds, 0.0181 mg for 7-15-year-olds, and ≥0.0188 mg for 16-year-olds. The mean exposure to egg protein under the low-intake scenario was estimated to be 0.0018 mg for 1-15-year-olds, 0.0019 mg for 4-15-year-olds, 0.0020 mg for 7-15-year-olds, and ≥0.0022 mg for 16-year-olds. Compared to the reference dose of 2.0 mg proposed by the Joint the Food and Agriculture Organization (FAO)/World Health Organization (WHO) Expert Committee, the risk of onset of food allergies due to egg protein contamination from foods without egg labeling is considered to be extremely low under the current Japanese food labeling system.

Structural modifications and augmented affinity for bile salts in enzymatically denatured egg white.

Protein binding to bile salts (BSs) reduces cholesterol levels, but the exact mechanism is unclear. In this study, we performed simulated gastrointestinal digestion of egg white protein hydrolysate (EWPHs) and included an unenzyme digestion group (CK) to investigate the changes in BSs binding capacity before and after digestion, as well as the relationship between egg white protein (EWP) structure and BSs binding capacity. In addition, peptidomics and molecular docking were used to clarify EWP's binding mechanism. We found that the BSs binding ability of EWPHs was slightly decreased after digestion, but significantly higher than that of the CK group and the digested CK group (D-CK). Particle size analysis and electrophoresis demonstrated that smaller particles and lower molecular weights exhibited enhanced binding capacity to BSs. Fourier Transform infrared spectroscopy (FTIR) results revealed that a disordered structure favored BS binding ability enhancement. Peptides FVLPM and GGGVW displayed hypocholesterolemic efficacy.

Effect of various contraceptives on oocyte yield and maturation in patients undergoing planned oocyte cryopreservation.

RESEARCH QUESTION: Do the various forms of hormonal and non-hormonal contraceptives have any association with ovarian stimulation outcomes, such as oocyte yield and maturation, in patients undergoing planned oocyte cryopreservation (POC)? DESIGN: This retrospective cohort study included all patients who underwent POC cycles between 2011 and 2023. The use of types of contraception before a POC cycle was recorded. The study evaluated the median number of cumulus-oocyte complexes obtained after vaginal oocyte retrieval and the proportion of metaphase II oocytes that underwent vitrification among all the cohorts. RESULTS: A total of 4059 oocyte freezing cycles were included in the analysis. Eight types of contraceptive method were recognized in patients undergoing ovarian stimulation: intrauterine device (IUD), copper (n = 84); IUD, levonorgestrel low dose (<52 mg) (n = 37); IUD, levonorgestrel (n = 192); subdermal etonogestrel implant (n = 14); injectable medroxyprogesterone acetate (n = 11); etonogestrel vaginal ring (n = 142); combined oral contraceptive pills (n = 2349); and norelgestromin transdermal patch (n = 10). The control group included patients not using contraceptives or using barrier or calendar methods (n = 1220). Among all the cohorts the median number of cumulus-oocyte complexes retrieved during oocyte retrieval was comparable (P = 0.054), and a significant difference in oocyte maturity rate with median number of vitrified oocytes was found (P = 0.03, P < 0.001, respectively). After adjusting for confounders a multivariate analysis found no association between the type of contraceptive and proportion of metaphase II oocytes available for cryopreservation. CONCLUSIONS: Among the various forms of contraception, none was shown to have an adverse association with oocyte yield or maturation rate in patients undergoing POC.

Effect of combined electrolyzed reduced water and slightly acidic electrolyzed water spraying on the control of Salmonella, eggshell quality, and shelf life of eggs during storage.

Slightly acidic electrolyzed water (SAEW) is a safe and effective disinfectant, but its sterilizing efficiency is compromised by organic matter on the egg surface. Electrolyzed reduced water (ERW) is a harmless cleaner with a decontamination effect on a variety of surfaces and can be used to remove organic matter. This study assesses the effectiveness of a combination of ERW and SAEW in eliminating Salmonella and manure mixture from egg surfaces, as well as its impact on egg quality during storage. The results show that ERW (74.14%) was more effective than deionized water (DW, 64.69%) and SAEW (70.20%) (P < 0.05) in removing manure from egg surfaces. The damage to the cuticle of eggshell treated with ERW for 28 s was similar to that of DW (P > 0.05) and less than that of SAEW (P < 0.05). Spraying ERW for 10 s followed by SAEW for 18 s (ERW + SAEW) completely removed Salmonella from the egg surface, with no bacteria detected in the residual wash solution. Additionally, ERW + SAEW demonstrated superior preservation of egg quality during storage at 25℃ than the control and ERW single treatment (P < 0.05). Moreover, ERW + SAEW resulted in less weight loss compared to SAEW single treatment (P < 0.05). In conclusion, the sequential use of ERW and SAEW appears to be a promising approach for sterilizing eggs. It not only removes organic matter and Salmonella from the egg surface but also improves the preservation quality of the egg at 25 ℃.

Sequence variations and accessory proteins adapt TMC functions to distinct sensory modalities.

Transmembrane channel-like (TMC) proteins are expressed throughout the animal kingdom and are thought to encode components of ion channels. Mammals express eight TMCs (mTMC1-8), two of which (mTMC1 and mTMC2) are subunits of mechanotransduction channels. C. elegans expresses two TMCs (TMC-1 and TMC-2), which mediate mechanosensation, egg laying, and alkaline sensing. The mechanisms by which nematode TMCs contribute to such diverse physiological processes and their functional relationship to mammalian mTMCs is unclear. Here, we show that association with accessory proteins tunes nematode TMC-1 to divergent sensory functions. In addition, distinct TMC-1 domains enable touch and alkaline sensing. Strikingly, these domains are segregated in mammals between mTMC1 and mTMC3. Consistent with these findings, mammalian mTMC1 can mediate mechanosensation in nematodes, while mTMC3 can mediate alkaline sensation. We conclude that sequence diversification and association with accessory proteins has led to the emergence of TMC protein complexes with diverse properties and physiological functions.

Macroscopic and microscopic morphological characteristics of gelatinous egg masses (egg veils) of yellow goosefish Lophius litulon (Lophiiformes; Lophiidae).

We recorded the morphological characteristics and effect of preservation methods on the structure of the egg veils of Lophius litulon found in field investigations. The egg veils were characterized as translucent sheet-shape with parallel opaque creases spaced approximately 2 cm apart. The egg veils were found to be composed of pentagonal or hexagonal chambers with rounded corners arranged in one layer, and each chamber enveloped one to three embryos. Cryopreservation is recommended to prevent structural changes in the egg veil rather than ethanol solution and neutral buffered formalin solution.

A comprehensive in-vitro/in-vivo screening toolbox for the elucidation of glucose homeostasis modulating properties of plant extracts (from roots) and its bioactives.

Plant extracts are increasingly recognized for their potential in modulating (postprandial) blood glucose levels. In this context, root extracts are of particular interest due to their high concentrations and often unique spectrum of plant bioactives. To identify new plant species with potential glucose-lowering activity, simple and robust methodologies are often required. For this narrative review, literature was sourced from scientific databases (primarily PubMed) in the period from June 2022 to January 2024. The regulatory targets of glucose homeostasis that could be modulated by bioactive plant compounds were used as search terms, either alone or in combination with the keyword "root extract". As a result, we present a comprehensive methodological toolbox for studying the glucose homeostasis modulating properties of plant extracts and its constituents. The described assays encompass in-vitro investigations involving enzyme inhibition (α-amylase, α-glucosidase, dipeptidyl peptidase 4), assessment of sodium-dependent glucose transporter 1 activity, and evaluation of glucose transporter 4 translocation. Furthermore, we describe a patch-clamp technique to assess the impact of extracts on KATP channels. While validating in-vitro findings in living organisms is imperative, we introduce two screenable in-vivo models (the hen's egg test and Drosophila melanogaster). Given that evaluation of the bioactivity of plant extracts in rodents and humans represents the current gold standard, we include approaches addressing this aspect. In summary, this review offers a systematic guide for screening plant extracts regarding their influence on key regulatory elements of glucose homeostasis, culminating in the assessment of their potential efficacy in-vivo. Moreover, application of the presented toolbox might contribute to further close the knowledge gap on the precise mechanisms of action of plant-derived compounds.

Impact of NaCl perturbation on physicochemical and structural properties of preheat-treated egg white protein modulating foaming property.

This study aimed to investigate the mechanism of NaCl perturbed preheat-treated egg white proteins' (EWPs) physicochemical and structural properties to modulate the foaming property (FP). The results revealed that NaCl regulated the salinolysis (5 mM) - salt precipitation (50 mM) - gradual or complete coverage with hydrated Na+ of the hydration layer (100-300 mM) - enhanced Cl- hydration repulsion (500 mM) of EWP, showing a gradual decrease in aggregates particle size, and reversibility of structural freedom, including moleculer flexibility and surface hydrophobicity. Whereas preheating temperature affected the secondary structure rearrangement and tertiary conformation exposure, and excessive temperature reduced foaming capacity while enhanced foam stability, with a tight correlation between NaCl-mediated EWPs' FP and the extent of Na+ covering the hydration layer. The findings provide a theoretical basis for processing factors to modulate the protein hydration layer to influence the functional properties.

Prevalence and determinants of food allergy in the era of early allergen introduction: the EarlyNuts population-based study.

BACKGROUND: Infant feeding guidelines in Australia changed in 2016 to recommend introduction of common allergy causing foods by age 1 year to prevent food allergy. Although the majority of Australian infants now eat peanut and egg by age 6-months, some still develop food allergy despite introducing allergens early. OBJECTIVE: We aimed to describe the prevalence of food allergy in a cohort recruited after introducing the nation-wide allergy prevention recommendations; identify characteristics of infants who developed allergy despite early introduction of allergens; and estimate the causal effect of modifiable exposures on food allergy prevalence and whether this differed between infants who were introduced to allergen before or after age 6 months. METHODS: We recruited a population-based sample of 12-month-old infants in Melbourne, Australia. Infants had skin prick tests to 4 foods and parents completed questionnaires. Infants with evidence of sensitisation were offered oral food challenges. Prevalence estimates were adjusted using inverse probability weighting. RESULTS: In a cohort of infants (n=1420) where nearly all infants had been introduced to common allergens such as egg, milk and peanut by one-year-of age, the prevalence of food allergy remained high at 11.3% (95% CI 9.6-13.4%). Infants who developed food allergy despite introducing the allergen by age 6-months were more likely to have Asian-born parents. Early-onset moderate/severe eczema was associated with an increased odds of food allergy, irrespective of whether allergens were introduced before or after age 6 months. Among infants who were introduced to peanut ≤6m, antibiotic use by age 6 months was associated with an increased odds of peanut allergy (aOR 6.03 (95%CI 1.15-31.60). CONCLUSION: In a cohort where early allergen introduction was common, the prevalence of food allergy remained high. Infants who developed food allergy despite introducing the respective allergen by 6 months were more likely to have Asian parents and early-onset eczema. New interventions are needed for infants with a phenotype of food allergy that is not amenable to early allergen introduction.

Biocontrol potential of endophytic fungi against phytopathogenic nematodes on potato (Solanum tuberosum L.).

Root-knot nematodes (RKNs) are a vital pest that causes significant yield losses and economic damage to potato plants. The use of chemical pesticides to control these nematodes has led to environmental concerns and the development of resistance in the nematode populations. Endophytic fungi offer an eco-friendly alternative to control these pests and produce secondary metabolites that have nematicidal activity against RKNs. The objective of this study is to assess the efficacy of Aspergillus flavus (ON146363), an entophyte fungus isolated from Trigonella foenum-graecum seeds, against Meloidogyne incognita in filtered culture broth using GC-MS analysis. Among them, various nematicidal secondary metabolites were produced: Gadoleic acid, Oleic acid di-ethanolamide, Oleic acid, and Palmitic acid. In addition, biochemical compounds such as Gallic acid, Catechin, Protocatechuic acid, Esculatin, Vanillic acid, Pyrocatechol, Coumarine, Cinnamic acid, 4, 3-indol butyl acetic acid and Naphthyl acetic acid by HPLC. The fungus was identified through morphological and molecular analysis, including ITS 1-4 regions of ribosomal DNA. In vitro experiments showed that culture filtrate of A. flavus had a variable effect on reducing the number of egg hatchings and larval mortality, with higher concentrations showing greater efficacy than Abamectin. The fungus inhibited the development and multiplication of M. incognita in potato plants, reducing the number of galls and eggs by 90% and 89%, respectively. A. flavus increased the activity of defense-related enzymes Chitinas, Catalyse, and Peroxidase after 15, 45, and 60 days. Leaching of the concentrated culture significantly reduced the second juveniles' stage to 97% /250 g soil and decreased the penetration of nematodes into the roots. A. flavus cultural filtrates via soil spraying improved seedling growth and reduced nematode propagation, resulting in systemic resistance to nematode infection. Therefore, A. flavus can be an effective biological control agent for root-knot nematodes in potato plants. This approach provides a sustainable solution for farmers and minimizes the environmental impact.