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This study aimed to evaluate the effects of herbicide 2, 4-D-dichlorophenoxy acetic acid on golden apple snail eggs and embryos. Additionally, the study assessed the applicability of optical coherence tomography (OCT), a non-invasive depth cross-sectional microscopic imaging technique, as a novel method, to the best of our knowledge, for studying morphological changes in golden apple snail eggs and embryos, in comparison to the conventional approach of using white light microscopy. The study revealed that the herbicide 2,4-D-dichlorophenoxy acetic acid affected the hatchery rate and morphological changes of the eggs and embryos. The lethal concentration (LC50), representing the concentration of a substance that is expected to cause death in half of the population being studied, of the golden apple eggs and embryos increased with longer exposure time and higher concentrations. The estimated median effective concentration (EC50), which denotes the concentration producing the desired effect in 50% of the exposed golden apple embryos, exhibited a similar trend of change as the LC50. When compared to the microscopic study, it was observed that OCT could be employed to investigate morphological changes of golden apple snail eggs and embryos, enabling evaluation of alterations in both 2D and 3D structures.

This study aimed to explore the effects of selenized glucose (SeGlu) and Na selenite supplementation on various aspects of laying hens such as production performance, egg quality, egg Se concentration, microbial population, antioxidant enzymes activity, immunological response, and yolk fatty acid profile. Using a 2 × 2 factorial design, 168 laying hens at 27-wk of age were randomly divided into 4 treatment groups with 7 replications. Se source (Na selenite and SeGlu) and Se level (0.3 and 0.6 mg/kg) were used as treatments. When 0.3 mg SeGlu/kg was compared to 0.3 mg Na selenite/kg, the interaction findings revealed that 0.3 mg SeGlu/kg increased egg production percent and shell ash (P < 0.05). When compared to 0.3 mg Na selenite/kg, dietary supplementation with 0.3 and 0.6 mg SeGlu/kg resulted in an increase in albumen height, Haugh unit, and yolk color of fresh eggs (P < 0.05). SeGlu enhanced albumen height, Haugh unit, shell thickness (P < 0.01), albumen index, yolk share, specific gravity, shell ash (P < 0.05) of fresh eggs and shell thickness (P < 0.05) of stored eggs as compared to Na selenite. The interaction showed that 0.6 mg SeGlu/kg enhanced yolk Se concentration while decreasing malondialdehyde levels in fresh egg yolk (P < 0.05). SeGlu enhanced Se concentration in albumen and glutathione peroxidase activity in plasma (P < 0.05) as compared to Na selenite. 0.6 mg Se/kg increased lactic acid bacteria, antibody response to sheep red blood cells, and lowered ∑n-6 PUFA/ ∑n-3 PUFA ratio (P < 0.05). As a result, adding SeGlu to the feed of laying hens enhanced egg production, egg quality, egg Se concentration, fresh yolk lipid oxidation, and glutathione peroxidase enzyme activity., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

The comet assay allows the analysis of DNA damage caused by different genotoxins. This assay has recently gained interest because of its ease of studying the interactions of xenobiotics with different organisms. Chrysoperla externa (Hagen, 1861) is a species of great economic relevance because it is a predator of major agricultural pests during its larval stage. Neonicotinoids are the most important chemical class of insecticides introduced into markets. A previous imidacloprid toxicity assessment on C. externa showed that this neonicotinoid insecticide reduced the egg viability. The objective of this study was to analyze the genotoxicity of Confidor OD® (imidacloprid 20% a.i., LS, Bayer CropScience) on the biological control agent C. externa at DNA level using the comet assay as an ecotoxicological biomarker. A comet assay protocol has been developed for this species at first time. For the bioassays, the commercial product formulated Confidor OD® was used at two concentrations: 100 and 180 mg/l of the active ingredient. Selected eggs were dipped in a Confidor OD® solution for 15 s. Descriptors evaluated in the comet assay were damage index, % DNA damage, and tail length. The damage index did not show any significant differences between the different concentrations evaluated, but differences were observed for tail length, because at higher concentrations of Confidor OD®, there were greater DNA breaks. The DNA of the cells from treated eggs analyzed at 48 h and 96 h of development showed the same % DNA damage; that is, they had no recovery capacity. Application of Confidor OD® to C. externa eggs produced irreparable breaks at the DNA level. The technique adjusted for C. externa can be used in other beneficial insects to study pesticide genotoxicity using a comet assay., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Marcela Ines Schneider reports financial support was provided by National Scientific and Technical Research Council., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

The aim of this study was to evaluate the effect of microencapsulated essential oils (MEO) on the laying performance, egg quality, immunity, intestinal morphology, and oxidative status of laying hens. A total of 640 Hy-line Brown laying hens, 41 wk of age, were randomly divided into 4 groups, each with 8 replicates containing 20 birds per replicate. The dietary conditions tested included a basal diet (Control) or the basal diet supplemented with various levels of MEO at 100 mg/kg (MEO100), 300 mg/kg (MEO300), and 500 mg/kg (MEO500). The three treatment groups were intermittently fed MEO, following an alternating schedule of 1 wk on and 1 wk off for a total of 56 d. Results showed that feeding MEO at levels of 300 and 500 mg/kg improved both egg production and feed conversion ratios compared to the control group. Hens consumed MEO-supplemented diets exhibited a significant decrease in the breaking egg ratio (P < 0.05) compared to those fed the control diet. Shell thickness and Haugh unit values significantly increased in the groups receiving 300 and 500 mg/kg of MEO (P < 0.05). Both the MEO300 and MEO500 treatments led to improvements in immunoglobulin (IgA, IgM, and IgG) and cytokine (IL-2 and IFN-γ) levels in serum. Hens in the MEO300 and MEO500 groups exhibited higher values for parameters related to intestinal morphometry compared to the control group. Furthermore, supplementation with 300 and 500 mg/kg of MEO enhanced the antioxidant capacity of plasma, as evidenced by increased activities of glutathione peroxidase (GSH-Px), total superoxide dismutase (T-SOD), and catalase (CAT) (P < 0.05). In summary, the intermittent feeding of MEO improved egg production, enhanced antioxidative processes, immune functions, and intestinal morphology, leading to an amelioration in the egg quality of laying hens. Our data demonstrate that supplementation of 300 mg/kg of MEO in feed can significantly improve animal health and egg quality. Implementation of these feeding practices could have a positive economic impact on poultry and egg industry., (Copyright © 2024. Published by Elsevier Inc.)

The aim of this study was to investigate the effects of puerarin (Pue), a phytoestrogen, on the production performance, egg quality, endocrine hormones, antioxidant capacity, and intestinal morphology in aged laying hens. A total of 180 Hy-Line Brown hens aged 480 d were randomly divided into 4 groups (n = 45 per group) and fed 0, 200, 400, and 800 mg/kg of Pue (Con, L-Pue, M-Pue, and H-Pue, respectively) during a 42-d experiment. Compared with the Con treatment, supplementation with H-Pue improved laying performance and egg quality by significantly increasing egg production, average egg weight, albumen height, yolk weight, and Haugh unit (P < 0.05) while decreasing the feed conversion ratio (P < 0.05). A diet supplemented with H-Pue significantly decreasing serum total triglycerides, total cholesterol, and low-density lipoprotein cholesterol, alanine aminotransferase (P < 0.05), and significantly increasing serum levels of follicle-stimulating hormone, luteinizing hormone and progesterone (P < 0.05). Antioxidant activity was improved by significantly increasing the activity of total antioxidant capacity, glutathione peroxidase and catalase but decreasing malondialdehyde levels in serum, jejunum, and ileum (P < 0.05), and superoxide dismutase activity exhibited a significantly increase in the jejunum and ileum (P < 0.05). Villus height and the ratio of villus height to crypt depth (P < 0.05) were significantly increased in the jejunum and ileum. In the jejunal and ileal mucosa, the three treatment groups increased the mRNA expression levels of Claudin-1 and Claudin-2 compared with Con (P < 0.05), and no significant effect was observed on the expression of Occludin and ZO-1. The results showed that dietary supplementation with Pue could improve the laying performance, egg quality, antioxidant capacity, hormonal profile, and intestinal morphology of aged laying hens., (Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)

Water pollution in developing countries continues to be a major health problem due to various anthropological activities that contribute to the spread of many parasitic diseases, including those caused by helminths. The aim of this study is to explore the ability of ozone and peroxone to disinfect drinking water contaminated samples with Toxocara canis eggs. The oxidants used were ozone and ozone-hydrogen peroxide combination. The treatment of Toxocara canis eggs was carried out in a 50 ml reactor with an operating volume of 10 ml. The pH conditions (5, 7 and 10) were varied for each treatment. The treatment effect was calculated by counting eggs and examining the condition of the larvae larval condition (whole, broken and hatched larvae) using an optical microscope. The experiment was carried out by exposing the eggs for 60 and 120 minutes to ozone and peroxone. The best results were obtained for helminths treated with the ozone/hydrogen peroxide combination at pH 10, with an inactivation of 79.2%. The synergistic effect of ozone combined with hydrogen peroxide allows higher helminth egg inactivation rates, demonstrating that advanced oxidation processes are a real alternative to apply in the inactivation of Toxocara canis eggs. The results obtained in this study show that the ozone and peroxone treatment could be a useful disinfection process to destroy or inactivate Toxocara canis eggs in processes commonly applied in water treatment.

Purpose: The purpose of this study was to evaluate the effectiveness of Pisonia alba Span commonly known as lettuce tree or bird catcher tree leaf extracts in controlling Aedes aegypti and Culex quinquefasciatus mosquitoes, which transmit several severe illnesses, such as dengue, Zika, chikungunya, yellow fever, West Nile virus, lymphatic filariasis and Japanese encephalitis., Methods: Sequential extraction of P. alba leaves was done using n-hexane, chloroform, and methanol. The larvicidal, pupicidal, and ovicidal activities of the extracts were evaluated against both mosquito species using standard bioassays. The concentrations of the extracts required to kill 50% of the mosquito larvae, pupae, and ova (LC 50 ) were determined., Results: The results showed that the methanol extract of P. alba had significant potential in killing the larvae, pupae, and ova of both mosquito species, with LC 50 values of 65.0 and 54.5 ppm for larvicidal activity, 73.7 and 69.7 ppm for pupicidal activity, and 78.4 and 80.0 ppm for ovicidal activity against Ae. aegypti and Cx. quinquefasciatus, respectively. The n-hexane extract demonstrated moderate larvicidal activity, while the n-hexane and chloroform extracts lacked ovicidal activity., Conclusion: The methanol extract from Pisonia alba could be a natural and eco-friendly alternative for controlling mosquito populations responsible for transmitting vector-borne diseases. Further research is warranted to determine their practical applications in controlling mosquito populations in the field., (© 2023. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Purpose: Ticks infestation has a negative impact against human and animal health through blood sucking, transmission of blood-borne diseases and also caused economic losses., Methods: In the present study the adulticidal, ovicidal and larvicidal activity of D-limonene nanoemulsion (DLN) were evaluated against two tick species; Rhipicephalus annulatus and Rhipicephalus sanguineus. Nanoemulsion form of D-limonene was prepared, and its characteristics were evaluated using a UV spectrophotometer and zeta droplet size measurement. Acetylcholinesterase activity was determined., Results: The results revealed significant adulticidal effect with low LC50 and LC90 for D-limonene pure form (DL) against both adult tick spp. (R. annulatus and R. sanguineus) ((0.958 and 1.559%) and (2.26 and 3.51%), respectively). DLN LC50 and LC90 values were ((1.277 and 2.396) and (3.97 and 7.28), respectively) against R. annulatus and R. sanguineus, respectively. DL and DLN showed significant ovicidal effect against R. sanguineus at high concentrations (10 and 5%). In larval packet test, LC50 and LC90 values of DL were ((1.53 and 2.22%) and (6.81 and 12.07%), respectively) against R. annulatus and R. sanguineus, respectively, while LC50 and LC90 values of DLN were ((6.48 and 11.26%) and (7.82 and 13.59%), respectively) against R. annulatus and R. sanguineus, respectively. Significant acetylcholinesterase inhibition percentage was detected for both ticks spp. which treated by DL and DLN., Conclusion: Pure DL is more effective than DLN form against R. annulatus and R. sanguineus., (© 2023. The Author(s) under exclusive licence to Witold Stefański Institute of Parasitology, Polish Academy of Sciences.)

Background: The use of insufficiently treated wastewater or Faecal sludge in agriculture raises concerns because of the pathogen content. Helminth eggs (HE) are one of the most crucial pathogens for ensuring public health and safety. Widely used disinfection treatment methods do not guarantee the complete inactivation of helminth eggs. The current study evaluated the effectiveness of anaerobic digestion and electrochemical process on helminth ( Ascaris suum ) egg inactivation., Methods: Lab-scale biochemical methane potential (BMP) assay was conducted by spiking A. suum eggs in a serum bottle. Total solid (TS), volatile solid (VS), pH, biogas production and its composition, and volatile fatty acids (VFA) were analyzed along with A. suum inactivation every third day for the initial 15 days and fifth day for 45 days. In the second set of experiments, a hypochlorite (4700 ppm) solution was generated by electrolysis of aqueous NaCl solution in a membrane-less electrochemical cell. The hypochlorite was diluted (940, 470, 235, and 156ppm) in wastewater, spiked with A. suum eggs and then examined for inactivation at regular intervals., Results: The results of the anaerobic digestion treatment documented 98% inactivation of A. suum eggs (0.15 eggs/mL) in 35 days and remained at 0.14 eggs/mL until day 45. Correlation analysis revealed a positive relationship between non-viable eggs and pH and a negative relationship with all the other parameters. Electrochemical treatment achieved 10% inactivation at 940 ppm concentration in 24h., Conclusions: This study revealed that the inactivation of A. suum eggs by anaerobic digestion or electrochemical treatment is a combined effect of more than one parameter., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 Patil PP and Mutnuri S.)

Chaya ( Cnidoscolus aconitifolius ) from family Euphorbiaceae is a plant has leaves similar to papaya leaves but with dark green leaf color. This plant contains phytochemical compounds such as alkaloid, flavonoid, triterpenoids, glycoside, and tannin that can function as antibacterial, antioxidant, and anti-inflammatory. Chaya leaf infusion are processed into infusion before given to the quails through drinking water, this method is used to obtain the phytochemical compounds contained in chaya leaves. This study aimed to evaluate chaya leaf infusion ( Cnidoscolus aconitifolius ) as a phytogenic source on the productivity and egg quality of japanese laying quail ( Coturnix coturnix japonica ) aged 17-20 weeks. Chaya leaf infusion is given through drinking water and is expected to increase the productivity and quality of quail egg. The variables observed consisted of productivity (water intake, feed intake, egg weigh, QDP, egg mass, FCR, mortality) and egg quality (yolk, albumen, shell weight and percentage, egg index, yolk index, albumen index, shell thickness, yolk color, and haugh unit). This study was analyzed using the T test with 2 treatments and 4 replications: T0 = drinking water without any additives and T1 = drinking water with a concentration 10% of chaya leaf infusion. The results showed that chaya leaf infusion had a significant effect ( P ≤0.05) on quail day production, egg mass, albumen index, and yolk color. Besides that, supplementation chaya leaf infusion significantly reduces ( P <0.05) feed convertion ratio. So that the use of 10% chaya leaf infusion in drinking water by giving it 2 times a week can increase the productivity and egg quality of laying quail ( Coturnix coturnix japonica ) 17-20 weeks old., Competing Interests: The authors declare that they have no conflict of interest.

This study investigated the effects of in ovo feeding (IOF) of l-arginine (L-Arg), selenium nanoparticles (SeNP), and a combination of L-Arg and SeNP on the hatchability, post-hatch growth, muscle development, antioxidant status, and meat quality of slow-growing chickens. On day 18 of incubation, a total of 960 fertilized eggs with similar weights were randomly assigned to 4 treatment groups with 4 replicates of 60 eggs each: (1) non-injected control group (Control), (2) injected with 1% of L-Arg (IOF&#95;L-Arg), (3) injected with 0.3 µg/egg of SeNP (IOF&#95;SeNP), and (4), injected with 1% of L-Arg and 0.3 µg/egg of SeNP (IOF&#95;L-Arg + SeNP). A completely randomized design was used. After hatching, 640 mixed-sex chicks were allocated to 4 treatment groups and split into 4 replicate pens (40 birds per pen). All groups of chicks were fed with commercial feed ad libitum until they reached 63 d of age and were subsequently weighed and slaughtered. The results of the present study showed that hatchability was similar among treatments. Final BW or breast muscle yield was not affected (P > 0.05) by IOF treatment. Chickens treated with IOF&#95;L-Arg + SeNP exhibited decreased feed conversion ratio, drip loss, and increased protein content in breast meat (P < 0.05). The IOF&#95;L-Arg + SeNP group exhibited a higher density of breast muscle fibers than the control group (P < 0.05). Overall, in ovo feeding of L-Arg combined with SeNP resulted in improved feed efficiency and enhanced antioxidant capacity at hatch without any adverse effects on chicken hatchability, health, or subsequent growth. Furthermore, meat from chickens in the IOF&#95;L-Arg + SeNP group exhibited a preferable texture with a higher protein content., (© The Author(s) 2024. Published by Oxford University Press on behalf of the American Society of Animal Science.)

Background Objectives: Insect growth regulators (IGRs) are biological hormone analogue or mimics used as pesticides to inhibit the growth of larva during their molting and skin shedding. This study aimed to test the effect of IGRs on the eggs hatching and post-hatching inhibition of Aedes mosquitoes and understanding its effect in the mosquito breeding habitats for reduction in adult emergence., Methods: Experiments on the evaluation of three insect growth regulators (IGRs) for the control of different stages of Aedes aegypti was carried out during 2020-21. Each experiment consisted of four treatments viz., Pyriproxyfen, Novaluron, and Larvicol at 1.0 ppm and distilled water as a control. All experiments were carried out in completely randomized design (CRD) except eggs which were carried out in factorial design each with three replications., Results: All tested IGRs performed better in affecting eggs, larval and pupal stages of Ae. aegypti. Highest eggs hatching inhibition (80%) of fresh eggs occurred in Pyriproxyfen followed by Novaluron (66%) and lowest in Larvicol (62%). Eggs hatch inhibition of embryonated eggs was lower than fresh eggs. Pyriproxyfen caused 69%, Novaluron 59% and Larvicol 39% eggs hatch inhibition of embryonated eggs. Both Pyriproxyfen and Novaluron performed better in causing 98-100% larval mortality followed by Larvicol (39%). Larval development to pupal stage was completely prevented by both Pyriproxyfen and Novaluron. Although Larvicol resulted in lowest eggs hatch and larval inhibition but prevented pupae to emerge as adults. Results further showed 70-89% mortality of 3rd instar larvae of Ae. aegypti when exposed to Pyriproxyfen and Novaluron solutions after 30 days storage at lab. temperature (27±2°C), RH 70±5., Interpretation Conclusion: None of the IGRs was more effective at the pupal stage but showed carry-on activity of growth inhibition and mortality of the successive stages of development when used against eggs stages. Therefore, we recommend early application of IGRs at mosquito habitats during the beginning and onset of the season when very early stages of mosquitoes are available in the field., (Copyright © 2024 Copyright: © 2024 Journal of Vector Borne Diseases.)

Cryoprotection is of interest in many fields of research, necessitating a greater understanding of different cryoprotective agents. Antifreeze proteins have been identified that have the ability to confer cryoprotection in certain organisms. Antifreeze proteins are an evolutionary adaptation that contributes to the freeze resistance of certain fish, insects, bacteria and plants. These proteins adsorb to an ice crystal's surface and restrict its growth within a certain temperature range. We investigated the ability of an antifreeze protein from the desert beetle Anatolica polita, ApAFP752, to confer cryoprotection in the frog Xenopus laevis. Xenopus laevis eggs and embryos microinjected with ApAFP752 exhibited reduced damage and increased survival after a freeze-thaw cycle in a concentration-dependent manner. We also demonstrate that ApAFP752 localizes to the plasma membrane in eggs and embryonic blastomeres and is not toxic for early development. These studies show the potential of an insect antifreeze protein to confer cryoprotection in amphibian eggs and embryos., Competing Interests: Competing interests The authors declare no competing or financial interests., (© 2022. Published by The Company of Biologists Ltd.)

Although employing nanocarriers for gene/drug delivery shows great potential in agricultural fields, the biotoxicity of nanocarriers is a major concern for large-scale applications. Herein, we synthesized a cationic star polymer (SPc) as a pesticide nanocarrier/adjuvant to evaluate its safety against a widely used predatory ladybird ( Harmonia axyridis ). The application of SPc at extremely high concentrations nearly did not influence the hatching of ladybird eggs but it led to the death of ladybird larvae at lethal concentration 50 (LC 50 ) values of 43.96 and 19.85 mg/mL through the soaking and feeding methods, respectively. The oral feeding of SPc downregulated many membrane protein genes and lysosome genes significantly, and the cell membrane and nucleus in gut tissues were remarkably damaged by SPc application, revealing that the lethal mechanism might be SPc-mediated membrane damage. Furthermore, the oral feeding of SPc increased the relative abundance of Serratia bacteria in ladybird guts to result in bacterial infection. Coapplication of ladybird and SPc-loaded thiamethoxam/matrine achieved desired control efficacies of more than 80% against green peach aphids, revealing that the coapplication could overcome the slow-acting property of ladybirds. To our knowledge, this is the first attempt to investigate the polymer-mediated lethal mechanism toward natural enemies and explore the possibility of coapplying SPc-loaded pesticides and natural enemies for pest management.

Soil-transmitted helminths (STH) are important and widespread intestinal pathogens of humans and animals. It is presently unknown which inactivating procedures may be universally effective for safe transport, preservation, and disinfection of STH-contaminated specimens, and this lack of knowledge may expose laboratory staff to higher risk of laboratory-acquired infections (LAI's). There are limited data on the efficacy of commonly used disinfectants and fecal fixatives for inactivating the eggs of STH. This work tested five disinfectants for surface cleanup, four storage temperature conditions, and six transport/storage fixatives, to inactivate eggs of three species of STH of animal origin ( Ascaris suum "roundworm," Trichuris vulpis "whipworm" and Ancylostoma caninum "hookworm") as surrogates for human STH. Among disinfectants, exposure to 10% povidone-iodine for ≥5 min inactivated 100% of the three species tested, while 5 min exposure to 95% ethanol inactivated T. vulpis and A. caninum eggs. All of the fixatives tested had inactivation effects on A. caninum hookworm eggs within 24 h of exposure, except potassium dichromate, which required 48 h. 95% ethanol for ≥48 h inactivated eggs from all three STH species. Freezing at ≤-20°C for ≥24 h inactivated eggs of T. vulpis and A. caninum, but only freezing at -80°C for ≥24 h inactivated >99% eggs, including A. suum . This work provides an evidence base for health and safety guidelines and mitigation strategies for the handling, storage, and disposal of stool samples containing STH eggs in laboratory, health care, childcare, or veterinary settings. IMPORTANCE This study systematically evaluates common laboratory disinfectants and storage conditions for their effectiveness in inactivating the infective stages of soil-transmitted helminths (STH). Animal-infecting proxy species were chosen to represent three major groups of STH that infect humans: roundworms, whipworms, and hookworms. Previously published work in this area typically focuses on a particular inactivation method, either for a single STH species, or on a subset of closely related species. Because prediagnostic fecal specimens must be regarded as potentially infectious with a mix of species, such information may be of limited utility in a working laboratory. We provide a straightforward summary of storage and disinfection methods that can achieve complete inactivation across a range of STH species, which represents a significant advance for clinical, veterinary and research laboratory biosafety.

The vitelline layer (VL) of a sea urchin egg is an intricate meshwork of glycoproteins that intimately ensheathes the plasma membrane. During fertilization, the VL plays important roles. Firstly, the receptors for sperm reside on the VL. Secondly, following cortical granule exocytosis, the VL is elevated and transformed into the fertilization envelope (FE), owing to the assembly and crosslinking of the extruded materials. As these two crucial stages involve the VL, its alteration was expected to affect the fertilization process. In the present study, we addressed this question by mildly treating the eggs with a reducing agent, dithiothreitol (DTT). A brief pretreatment with DTT resulted in partial disruption of the VL, as judged by electron microscopy and by a novel fluorescent polyamine probe that selectively labelled the VL. The DTT-pretreated eggs did not elevate the FE but were mostly monospermic at fertilization. These eggs also manifested certain anomalies at fertilization: (i) compromised Ca 2+ signaling, (ii) blocked translocation of cortical actin filaments, and (iii) impaired cleavage. Some of these phenotypic changes were reversed by restoring the DTT-exposed eggs in normal seawater prior to fertilization. Our findings suggest that the FE is not the decisive factor preventing polyspermy and that the integrity of the VL is nonetheless crucial to the egg's fertilization response.

Background: The limited ovicidal activity of currently available acaricides is a significant obstacle to efficacious scabies treatment. Several essential oils or their respective components have proved to be active against the eggs of arthropods, mainly lice and ticks. Information on the activity of these oils and/or components against the eggs of mites remains very limited. The aim of this study was to assess the activity of six terpenes (carvacrol, eugenol, geraniol, citral, terpinen-4-ol and linalool) commonly found in essential oils against the eggs of Sarcoptes scabiei., Methods: Sarcoptes eggs were exposed to paraffin oil containing 1, 2.5, or 5% of each terpene tested. After a 12-h exposure period, the eggs were washed and placed in paraffin oil for hatching. Embryonic development following treatment was assessed every day to determine the stage of developmental arrest., Results: The median effective concentration to obtain 50% egg mortality (EC 50 ) was 0.5, 0.9, 2.0, 4.8, 5.1 and 9.8% for carvacrol, eugenol, geraniol, citral, terpinen-4-ol and linalool, respectively. The microscopic images of eggs after each treatment indicated that these six terpenes may act by penetrating through the aeropyles on the egg surface., Conclusions: In conclusion, carvacrol, eugenol and geraniol possess significant ovicidal activities, which should be considered as promising ovicidal agents for the treatment of scabies., (© 2021. The Author(s).)

Helminth infections remain a major constraint to livestock productivity in the tropical and subtropical areas across the world, especially in the areas where extensive grazing is practiced. The development of resistance to anthelmintic drugs, scarcity and high cost of purchase especially to farmers of low income in developing countries led to the need of alternative helminth control methods. However, there is an urgent need to discover novel drugs that can cure helminthiasis. A survey was carried out among agropastoralists to elicit information on the use of Ethnoveterinary Plants (EvPs) as alternative medicine for helminthiasis. The plants used by the agropastoralists were collected and identified at the Department of Botany Herbarium, University of Ibadan. The plant part was pulverized and cold macerated successively with n-hexane, ethyl acetate, methanol and aqueous methanol to obtain crude extracts. The methanolic extract was assayed against Haemonchus contortus at test concentrations (6.25, 12.5, 25, 50 and 100 µg/mL) and Albendazole at 25µg/mL were tested for the egg hatch inhibition assay. Eggs hatched and unhatched were counted under the microscope at 48, 96 and 144 hours (h). Among the EvPs identified, Terminalia glaucescens was the least utilized plant by the agropastoralists, and thus, selected for evaluation. The preliminary phytochemical screening revealed presence of tannin, alkaloid, saponin, flavonoid, phenolic, steroids, glycosides, triterpenes and reducing sugars. At 96 h, there were significant difference (P<0.05) in egg hatch inhibition (EHI) percentage at 100ug/mL (87.55), 50µg/mL (84.29) at inhibitory concentration (IC 50 value 1.07) compared to 25ug/mL (100) for the Albendazol. At 144 h, there were no significant (P>0.05) differences observed in EHI values of methanolic extract of the leaf at 100ug/mL (89.69), 50ug/mL (87.06), 25ug/mL (85.53) and 12.5µg/mL (82.89) at IC 50 value 1.08 compared to 25ug/mL (100) for the Albendazol. T. glaucescens leaf is a potential source of novel anthelmintics and further investigation should be carried out on its in vivo anthelmintic activity., (Copyright © 2021. Published by Elsevier B.V.)

In the present study, an anthelmintic treatment regimen with reduced treatment frequency was evaluated in horses on two study sites in Belgium during three consecutive summer pasture seasons. Historically, the horses on both study sites were treated up to 6 times a year with ivermectin (IVM) or up to 4 times a year with moxidectin (MOX), and previous efficacy evaluations indicated a reduced egg reappearance period in some of the treated horses for both IVM (28 days) and MOX (42 days). In the present study, all horses were treated with IVM or MOX in the spring and in autumn. Faecal egg counts (FEC) were conducted every two weeks during the summer pasture season and whenever the individual FEC exceeded 250 eggs per gram of faeces, the specific horse was treated with pyrantel embonate. No increase in parasitic disease over the three-year period of the study was observed. The FEC data collected in the study as well as the age of the animals and local weather data were then imported into a cyathostomin life-cycle model, to evaluate long term effects of the newly applied treatment regimen on the selection pressure for anthelmintic resistance, and compare to the previous high frequency treatment regimen. The model simulations indicated that the whole-herd treatment regimen with at least 4 macrocyclic lactone treatments annually led 2-3 times faster resistance development than any of the alternative treatment regimens evaluated under the specific conditions of these two study sites. Further lowering the treatment frequency or applying even more selective treatments enhanced the delay in resistance development, but to a lesser extent., (Copyright © 2021. Published by Elsevier B.V.)

Background: There is an urgent need for an extensive evaluation of benzimidazole efficacy in humans. In veterinary science, benzimidazole resistance has been mainly associated with three single-nucleotide polymorphisms (SNPs) in the isotype-1 β-tubulin gene. In this study, we optimized the stool sample processing methodology and resistance allele frequency assessment in Trichuris trichiura and Necator americanus anthelmintic-related SNPs by pyrosequencing, and standardized it for large-scale benzimidazole efficacy screening use., Methods: Three different protocols for stool sample processing were compared in 19 T. trichiura-positive samples: fresh stool, egg concentration using metallic sieves with decreasing pore size, and egg concentration followed by flotation with saturated salt solution. Yield of each protocol was assessed by estimating the load of parasite DNA by real-time PCR. Then, we sequenced a DNA fragment of the β-tubulin gene containing the putative benzimidazole resistance SNPs in T. trichiura and N. americanus. Afterwards, resistant and susceptible-type plasmids were produced and mixed at different proportions, simulating different resistance levels. These mixtures were used to compare previously described pyrosequencing assays with processes newly designed by our own group. Once the stool sample processing and the pyrosequencing methodology was defined, the utility of the protocols was assessed by measuring the frequencies of putative resistance SNPs in 15 T. trichiura- and 15 N. americanus-positive stool samples., Results: The highest DNA load was provided by egg concentration using metallic sieves with decreasing pore size. Sequencing information of the β-tubulin gene in Mozambican specimens was highly similar to the sequences previously reported, for T. trichiura and N. americanus, despite the origin of the sample. When we compared pyrosequencing assays using plasmids constructs, primers designed in this study provided the most accurate SNP frequencies. When pooled egg samples were analysed, none of resistant SNPs were observed in T. trichiura, whereas 17% of the resistant SNPs at codon 198 were found in one N. americanus sample., Conclusions: We optimized the sample processing methodology and standardized pyrosequencing in soil-transmitted helminth (STH) pooled eggs. These protocols could be used in STH large-scale screenings or anthelmintic efficacy trials., (© 2021. The Author(s).)

Itaconic acid is produced by immune responsive gene 1 (IRG1)-coded enzyme in activated macrophages and known to play an important role in metabolism and immunity. In this study, mechanism of itaconic acid functioning as an anti-inflammatory metabolite was investigated with molecular biology and immunology techniques, by employing IRG1-null (prepared with CRISPR) and wild-type macrophages. Experimental results showed that itaconic acid significantly promoted the pentose phosphate pathway (PPP), which subsequently led to significantly higher NADPH oxidase activity and more reactive oxygen species (ROS) production. ROS production increased the expression of anti-inflammatory gene A20, which in turn decreased the production of inflammatory cytokines IL-6, IL-1β and TNF-α. NF-κB, which can up-regulate A20, was also vital in controlling IRG1 and itaconic acid involved immune-modulatory responses in LPS-stimulated macrophage in this study. In addition, itaconic acid inhibited the growth of Salmonella typhimurium in cell through increasing ROS production from NADPH oxidase and the hatching of Schistosoma japonicum eggs in vitro. In short, this study revealed an alternative mechanism by which itaconic acid acts as an anti-inflammatory metabolite and confirmed the inhibition of bacterial pathogens with itaconic acid via ROS in cell. These findings provide the basic knowledge for future biological applications of itaconic acid in anti-inflammation and related pathogens control., (© 2021. The Author(s).)

Gastrointestinal nematodes (GIN) infections are a serious problem in livestock production due to the great economic losses they cause. Their control is increasingly difficult because of the rapid development of drug resistance and the limited number of available drugs. Therefore, this study evaluated 18 aminoalcohol and 16 diamine derivatives against eggs, first and third stage larvae from a susceptible and a resistant isolate of Teladorsagia circumcincta collected from sheep. The effectiveness of the in vitro anthelmintic activity of the compounds was evaluated using three different procedures: Egg Hatch Test (EHT), Larval Mortality Test (LMT) and Larval Migration Inhibition Test (LMIT). Those compounds with activities higher than 90 % in the initial screening at 50 μM were selected to determine their half maximal effective concentration (EC 50 ). In parallel, cytotoxicity assays were conducted on Caco2 and HepG2 cell lines to calculate Selectivity Indexes (SI) for each compound. The diamine 30 presented the best results in preventing egg hatching, displaying the lowest EC 50 value (1.01 ± 0.04 μM) of all compounds tested and the highest SI (21.21 vs. Caco-2 cells). For the LMIT, the diamine 34 showed the highest efficacy, with EC 50 values of 2.67 ± 0.08 and 3.02 ± 0.09 μM on the susceptible and resistant isolate of the parasite, respectively., (Copyright © 2021. Published by Elsevier B.V.)

Investigating the mechanisms of action of natural bioactive products against pests is a vital strategy to develop novel promising biopesticides. Scoparone, isolated from Artemisia capillaris, exhibited potent oviposition inhibition activity against Tetranychus cinnabarinus (a crop-threatening mite pests with strong fecundity). To explore the underlying mechanism, the vitellogenin (Vg) protein content, and Vg gene expression of mites from three consecutive generations of G0 individuals exposed to scoparone were determined, revealing marked inhibition. This study is the first to explore the egg development defect behaviour of mite pests induced by scoparone. The egg-laying inhibition of mites by scoparone was significantly increased by 47.43% compared with that of the control when TcVg was silenced by RNA interference (RNAi), suggesting that egg-development inhibition of female T. cinnabarinus by scoparone was mediated by low Vg gene expression. Furthermore, scoparone bound to the Vg protein in vitro, and its K d value was 218.9 μM, implying its potential function in inhibiting the egg development of mites by directly targeting the Vg protein. This study will lay the foundation for the future applications of scoparone as an agrochemical for controlling the strong egg-laying capacity mite pests in agriculture., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The tomato leaf miner (TLM), Tuta absoluta (Meyrick), is an invasive tomato pest found worldwide. Sustainable control strategies aimed at increasing biological control approaches and decreasing chemical inputs are required, due to the tendency to develop insecticide resistance. In this study, the lethal and sublethal effects of four chemical insecticides (abamectin, indoxacarb, chlorantraniliprole, and spinosad) and the sublethal effects of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) on a widespread TLM egg parasitoid, Trichogramma brassicae Bezdenko, were estimated. Concentration mortality response bioassays enabled the estimation of lethal concentrations of the tested insecticides for the parasitoids, with chlorantraniliprole having the lowest LC50 and indoxacarb the highest. The LC25 and LC50 of the tested insecticides on the TLM were sprayed on eggs and then offered at three time intervals to the parasitoids. The fertility and other life table parameters of the individuals emerging from the treated eggs were estimated. All of the chemical insecticides, but not the fungus, had harmful effects on T. brassicae. The insecticide applications caused a 3.84-5.17 times reduction in the net reproductive rate (R0) compared with the control. No parameters were affected by spraying the fungus in the 0h treatment, but effects were recorded at 24 and/or 48h, except for the gross reproduction rate (GRR). The value of the intrinsic rate of increase (rm) also decreased to 0.528-0.617 after the insecticide treatments. The doubling time (DT) increased in all treatments compared to the control. Nevertheless, the generation time (T) was only very slightly affected. In addition, in the combination experiments, M. anisopliae showed a remarkable synergism with T. brassicae in controlling TLM eggs. These results indicate that low levels of lethal effects on key biological control agents should be considered in the choice of insecticides to be included in sustainable TLM control packages., Competing Interests: The authors have declared that no competing interests exist.

Mammalian oocytes undergo major changes in zinc content and localization to be fertilized, the most striking being the rapid exocytosis of over 10 billion zinc ions in what are known as zinc sparks. Here, we report that fertilization of amphibian Xenopus laevis eggs also initiates a zinc spark that progresses across the cell surface in coordination with dynamic calcium waves. This zinc exocytosis is accompanied by a newly recognized loss of intracellular manganese. Synchrotron-based X-ray fluorescence and analytical electron microscopy reveal that zinc and manganese are sequestered in a system of cortical granules that are abundant at the animal pole. Through electron-nuclear double-resonance studies, we rule out Mn 2+ complexation with phosphate or nitrogenous ligands in intact eggs, but the data are consistent with a carboxylate coordination environment. Our observations suggest that zinc and manganese fluxes are a conserved feature of fertilization in vertebrates and that they function as part of a physiological block to polyspermy.

The bio-efficacy of crude ethyl acetate extract, fractions and a compound phenyl acetic acid from the ethyl acetate extract of Streptomyces collinus was evaluated on Culex quinquefasciatus Say and Aedes aegypti L. mosquitoes (Diptera: Culicidae). The larvae were exposed to concentrations of 2.5, 5.0, 7.5 and 10.0 ppm for fractions and 0.5, 1.0, 1.5 and 2.0 ppm for compound. After 24 h, the larval mortality was assessed and the LC₅₀ and LC 90 values were calculated. Similarly, per cent ovicidal activity was calculated for eggs after 120 h post treatment for phenyl acetic acid. Among the eleven fractions screened, fraction 7 from the ethyl acetate extract of Streptomyces collinus exhibited good larvicidal activity against both mosquito species. The LC 50 and LC 90 values of fraction 7 were 4.42, 6.23 ppm against Cx. quinquefasciatus larvae and 5.13, 14.51 ppm against Ae. aegypti larvae, respectively. Further, the isolated compound, phenyl acetic acid from fraction 7 recorded 100% larvicidal activity at 2 ppm concentration with LC 50 and LC 90 values of 2.07, 4.87 ppm on Cx. quinquefasciatus larvae and 3.81, 9.87 ppm on Ae. aegypti larvae, respectively. Phenyl acetic acid presented 50.3% and 42.0% ovicidal activity against Cx. quinquefasciatus and Ae. aegypti eggs at 2 ppm concentration after 120 h post treatment. The compound, phenyl acetic acid could be used in mosquito control programme., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Following the previous findings reported by the present authors on the anthelmintic effect of hydro-ethanolic extract of Mentha pulegium, the volatile constituents of M. pulegium are now assessed in the present study by exploring its anthelmintic and its antioxidant proprieties using in vitro and in vivo assays. Egg hatch assay (EHA) and adult worm's motility assays (AWMA) were used to assess the in vitro activity against Haemonchus. contortus. The in vivo anthelmintic potential was evaluated in mice infected with Heligmosomoides polygyrus using faecal egg count reduction (FECR) and total worm count reduction (TWCR). M. pulegium EO demonstrated 100% inhibition in the EHA at 200 μg/mL (IC50 = 56.36 μg/mL). In the AWM assay, EO achieved total worms paralysis 6 h after treatment exposure. This nematicidal effect was associated to morphological damages observed in the cuticular's worm using environmental scanning electron microscopy (ESEM). At 400 mg/kg, M. pulegium oil showed 75.66% of FECR and 80.23% of TWCR. The antioxidant potential of this plant was also monitored by several in vitro assays: total antioxidant capacity was 205.22 mg GAE/g DW, DPPH quenching effect was IC50 = 140 μg/mL, ABTS activity IC50 = 155 μg/mL and FRAP effect of 660 μg/mL. Regarding the in vivo assay, M. pulegium EO demonstrated a protective effect against oxidative stress by increasing the activity of the endogenous antioxidants (SOD, CAT and GPx) during H. polygyrus infection., (Copyright © 2021 Elsevier Inc. All rights reserved.)

Many bacteria, such as Pseudomonas aeruginosa, regulate phenotypic switching in a population density-dependent manner through a phenomenon known as quorum sensing (QS). For Gram-negative bacteria, QS relies on the synthesis, transmission, and perception of low-molecular-weight signal molecules that are predominantly N-acyl-l-homoserine lactones (AHLs). Efforts to disrupt AHL-mediated QS have largely focused on the development of synthetic AHL analogues (SAHLAs) that are structurally similar to native AHLs. However, like AHLs, these molecules tend to be hydrophobic and are poorly soluble under aqueous conditions. Water-soluble macrocycles, such as cyclodextrins (CDs), that encapsulate hydrophobic guests have long been used by both the agricultural and pharmaceutical industries to overcome the solubility issues associated with hydrophobic compounds of interest. Conveniently, CDs have also demonstrated anti-AHL-mediated QS effects. Here, using fluorescence spectroscopy, NMR spectrometry, and mass spectrometry, we evaluate the affinity of SAHLAs, as well as their hydrolysis products, for β-CD inclusion. We also evaluated the ability of these complexes to inhibit wild-type P. aeruginosa virulence in a Caenorhabditis elegans host infection study, for the first time. Our efforts confirm the potential of β-CDs for the improved delivery of SAHLAs at the host/microbial interface, expanding the utility of this approach as a strategy for probing and controlling QS., (© 2020 Wiley-VCH GmbH.)

The continuous use of synthetic anthelmintics against gastrointestinal nematodes (GINs) has resulted in the increased resistance, which is why alternative methods are being sought, such as the use of natural products. Plant essential oils (EOs) have been considered as potential products for the control of GINs. However, the chemical composition and, consequently, the biological activity of EOs vary in different plant cultivars. The aim of this study was to evaluate the anthelmintic activity of EOs from cultivars of Ocimum basilicum L. and that of their major constituents against Haemonchus contortus. The EOs from 16 cultivars as well the pure compound linalool, methyl chavicol, citral and eugenol were used in the assessment of the inhibition of H. contortus egg hatch. In addition, the composition of three cultivars was simulated using a combination of the two major compounds from each. The EOs from different cultivars showed mean Inhibition Concentration (IC50) varying from 0.56 to 2.22 mg/mL. The cultivar with the highest egg-hatch inhibition, Napoletano, is constituted mainly of linalool and methyl chavicol. Among the individual compounds tested, citral was the most effective (IC50 0.30 mg/mL). The best combination of compounds was obtained with 11% eugenol plus 64% linalool (IC50 0.44 mg/mL), simulating the Italian Large Leaf (Richters) cultivar. We conclude that different cultivars of O. basilicum show different anthelmintic potential, with cultivars containing linalool and methyl chavicol being the most promising; and that citral or methyl chavicol isolated should also be considered for the development of new anthelmintic formulations.

Background: Credelio™ (lotilaner; Elanco) is indicated for the treatment of flea and tick infestations on cats at a recommended lotilaner dose rate of 6-24 mg/kg. This study evaluated the efficacy and safety of lotilaner following a single oral administration to cats for the treatment and prevention of adult Ctenocephalides felis fleas and flea egg production under laboratory conditions., Methods: Two treatment groups of ten cats each were used in this study. One group was treated with lotilaner at a dose rate of 6-9 mg/kg on Day 0 and the other group served as the control group. Each cat was infested with 100 unfed adult fleas on days -1, 6, 13, 20 and 29. At 24 h post-treatment or post-infestation, each cat was combed to remove and count adult live fleas. At each time point, flea eggs were also collected and counted from under each cat cage., Results: Following a single oral administration of lotilaner at a minimum dose rate of 6 mg/kg (range 6.00-8.57 mg/kg), the lotilaner group displayed 100%, 100%, 99.9%, 99.9% and 99.8% efficacy against adult live flea counts as compared to the control group on Days 1, 7, 14, 21 and 30, respectively. At each time point, adult flea counts from the lotilaner-treated cats were significantly lower (P < 0.0001) than from the control group. A mean flea egg count of 22.6 in the lotilaner-treated cats (compared to 441.7 in the control animals) was observed 24 h post-treatment. No eggs were present from any of the treated cats on Days 7, 14 and 30 and a single egg was detected on a single treated cat on Day 21. One adverse event (regurgitated food) was observed during the study in one treated cat approximately 1 h after dosing., Conclusions: Lotilaner was well tolerated; only one adverse event was observed in the treated group. Virtually all adult fleas were killed within 24 h post-treatment or post-infestation in cats treated with a single dose of lotilaner as compared to the control group, thus significantly reducing the number of flea eggs being produced for 30 days after treatment.

Rationale: Accumulated evidence indicates that environmental plasticizers are a threat to human and animal fertility. Di (2-ethylhexyl) phthalate (DEHP), a plasticizer to which humans are exposed daily, can trigger reproductive toxicity by acting as an endocrine-disrupting chemical. In mammals, the female primordial follicle pool forms the lifetime available ovarian reserve, which does not undergo regeneration once it is established during the fetal and neonatal period. It is therefore critical to examine the toxicity of DEHP regarding the establishment of the ovarian reserve as it has not been well investigated. Methods: The ovarian cells of postnatal pups, following maternal DEHP exposure, were prepared for single cell-RNA sequencing, and the effects of DEHP on primordial follicle formation were revealed using gene differential expression analysis and single-cell developmental trajectory. In addition, further biochemical experiments, including immunohistochemical staining, apoptosis detection, and Western blotting, were performed to verify the dataset results. Results: Using single-cell RNA sequencing, we revealed the gene expression dynamics of female germ cells and granulosa cells following exposure to DEHP in mice. Regarding germ cells: DEHP impeded the progression of follicle assembly and interfered with their developmental status, while key genes such as Lhx8 , Figla, and others, strongly evidenced the reduction. As for granulosa cells: DEHP likely inhibited their proliferative activity, and activated the regulation of cell death. Furthermore, the interaction between ovarian cells mediated by transforming growth factor-beta signaling, was disrupted by DEHP exposure, since the expression of GDF9, BMPR1A, and SMAD3 was affected. In addition, DNA damage and apoptosis were elevated in germ cells and/or somatic cells. Conclusion: These findings offer substantial novel insights into the reproductive toxicity of DEHP exposure during murine germ cell cyst breakdown and primordial follicle formation. These results may enhance the understanding of DEHP exposure on reproductive health., Competing Interests: Competing Interests: The authors have declared that no competing interest exists., (© The author(s).)

Current risk assessment strategies for honey bees rely heavily upon laboratory tests performed on adult or immature worker bees, but these methods may not accurately capture the effects of agrochemical exposure on honey bee queens. As the sole producer of fertilized eggs inside a honeybee colony, the queen is arguably the most important single member of a functioning colony unit. Therefore, understanding how agrochemicals affect queen health and productivity should be considered a critical aspect of pesticide risk assessment. Here, an adapted method is presented to expose honey bee queens and worker queen attendants to agrochemical stressors administered through a worker diet, followed by tracking egg production in the laboratory and assessing first instar eclosion using a specialized cage, referred to as a Queen Monitoring Cage. To illustrate the method's intended use, results of an experiment in which worker queen attendants were fed diet containing sublethal doses of imidacloprid and effects on queens were monitored are described.

Emerging infectious diseases are major drivers of global and local amphibian biodiversity loss. Therefore, developing effective disinfection methods to manage the impact of diseases in wild and captive "ark" populations are an important goal in amphibian conservation. While chemical disinfectants have been used safely and effectively in larval and adult amphibians infected with pathogenic microbes, their applicability to amphibian egg masses has remained untested. To bridge this gap, we exposed embryos of the common toad (Bufo bufo) and agile frog (Rana dalmatina) experimentally to three widely used disinfectants: voriconazole, chloramphenicol and chlorogen-sesquihydrate. For 3 days we exposed portions of egg masses to these disinfectants at 1×, 2×, 5× and 10× the concentration recommended for the disinfection of tadpoles and adults. Subsequently, we recorded embryonic and larval survival, as well as larval body mass and the incidence of abnormalities 12 days after hatching. Application of voriconazole had species- and concentration-dependent negative impacts on survival and body mass, and caused marked malformations in the viscerocranial structure of B. bufo tadpoles. Exposure to chlorogen-sesquihydrate also resulted in significant mortality in B. bufo embryos and negatively affected body mass of R. dalmatina larvae. Chloramphenicol had little negative effects on embryos or larvae in either species. Based on these results, the application of voriconazole and chlorogen-sesquihydrate cannot be recommended for the disinfection of amphibian eggs, whereas treatment with chloramphenicol appears to be a safe method for eliminating potential pathogens from anuran egg masses and their immediate aquatic environment., (© 2020 John Wiley & Sons, Ltd.)

Nearly a century ago, studies by Lancelot Hogben and others demonstrated that ovulation in female Xenopus laevis can be induced via injection of mammalian gonadotropins into the dorsal lymph sac, allowing for egg production throughout the year independent of the normal reproductive cycles. Hormonally induced females are capable of producing thousands of eggs in a single spawning, which can then be fertilized to generate embryos or used as a substrate for generation of egg extracts. The protocol for induction of ovulation and subsequent egg collection is straightforward and robust, yet some of its details may vary among laboratories based on prior training, availability of necessary reagents, or the experimental objectives. As the goal of this protocol is not to describe every single variation possible for acquiring eggs but to provide a simple and clear description that can be easily applied by researchers with no prior working experience with X. laevis , we focus on describing the method we use at the National Xenopus Resource-that is, inducing ovulation in X. laevis via dorsal lymph sac injection of gonadotropic hormones and the stimulation of egg laying through application of gentle pressure to the females., (© 2021 Cold Spring Harbor Laboratory Press.)

The objective of this study was to investigate the effects of dietary copper (Cu) on production, egg quality, and hatchability of Chinese Yellow broiler breeder hens and growth performance of their offspring. A total of 576 30-week-old hens were randomly allotted into 6 groups, each with 6 replicates (8 cages for each replicate with 2 birds per cage). The basal diet contained 3.50 mg/kg Cu, and the other 5 treatment diets contained 8.5, 13.5, 23.5 43.5, and 83.5 mg/kg Cu, respectively, additionally supplemented with Cu on the basal diet. The trial lasted for 15 wk. Qualified egg rate of birds fed 23.5 or 83.5 mg/kg Cu was significantly decreased (P < 0.05) compared with those given 3.5, 8.5, or 13.5 mg/kg Cu. Plasma malondialdehyde concentration showed quadratic effect (P = 0.002) which that decreased first then increased with dietary Cu increased. Highest values of Cu content and hepatic activity of Cu-ATPase occurred in hens fed 83.5 mg/kg dietary Cu with linear (P = 0.001) and quadratic (P = 0.001) effects. Shell strength and proportion on 18th day of live embryos of hens fed 13.5 mg/kg Cu were the greatest compared with other groups respectively (P < 0.05); rate of qualified eggs for hatch and hatchability of fertilized eggs of hens fed 83.5 mg/kg Cu were the least (P < 0.05). In conclusion, both inadequate (3.5 mg/kg diet) and excess (83.5 mg/kg) of dietary Cu can induce oxidative stress in hens and lead to decreased egg quality. Hatchability and growth performance of offspring were decreased when breeder hens were fed excess Cu in spite of greater hatching weight. The appropriate dietary Cu level for Chinese Yellow broiler breeder hens during the egg-laying period is 15.7 to 21.2 mg/kg (1.81-2.44 mg Cu fed per day) when based on Cu level and Cu-ATPase activity in the liver. This dietary Cu requirement is approximately doubled (∼40 mg/kg, ∼4.60 mg Cu per bird per day) for maximal response of eggshell thickness., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The branched isomer mixture 4-nonylphenol (4-NP) has been used worldwide as a surfactant, and can have endocrine-disrupting effects on aquatic organisms. For instance, 4-NP induces the formation of testis-ova (i.e., testicular and ovarian tissue in the same gonad) or male to female sex reversal of various teleost fishes. Recently, our group revealed that altered gsdf gene expression is associated with disruption of gonadal differentiation in Japanese medaka (Oryzias latipes) embryos exposed to methyltestosterone or bisphenol A, suggesting that gsdf might be useful as a biomarker for predicting the impact of endocrine-disrupting chemicals (EDCs) on gonadal differentiation. Here, we used 4-NP to examine further whether gsdf expression at the embryo stage is useful for predicting EDC impact on gonadal sex differentiation. When fertilized medaka eggs were exposed to 32 or 100 μg/L 4-NP, testis-ova in genetic males and sex reversal from genetic male to phenotypic female were observed. At stage 38 (just before hatching), 4-NP exposure at 1-100 μg/L did not affect gsdf expression in XX embryos compared with the nontreated control; however, in XY embryos, the gsdf expression in the 100 μg/L-exposed group was significantly lower than that in the controls. The 4-NP concentration at which gsdf expression was suppressed was equal to that at which testis-ova and sex reversal were induced. These results indicate that expression of the gsdf gene at the embryonic stage in medaka is a useful biomarker for predicting the impact of EDCs on sexual differentiation., (© 2020 John Wiley & Sons, Ltd.)

To investigate the effects of replacing maize with various levels of raw and processed oak fruit in diet on productive performance of laying hens and egg quality traits, the total number of 168 Bovans white laying hens (30-week age) were randomly distributed between 28 replicate cages and assigned to 7 experimental diets. Based on a 2 × 3 factorial arrangement of treatments including two dietary levels (10% and 20%) of raw, soaked or boiled oak fruit as well as a corn-soybean meal-based diet, 7 experimental diets with 4 replicates and 6 chickens per replicate cages were evaluated during an 8-week period. The data were analysed using GLM procedure of SAS. Significantly higher feed consumption was observed in 10% boiled oak compared with soaked oak (p < .05). Significantly improved feed conversion ratio was observed in 10% boiled oak compared with soaked oak (p < .05). Diet inclusion of 10% oak fruit caused significant increased shell weight. Significant interaction between oak level and processing method on the egg-specific gravity was observed (p < .05), and higher egg-specific gravity was detected in hens fed the diets included 20% boiled or 20% raw oak compared to 20% soaked oak. Significantly increased blood LDL level was observed in hens fed the diets that included boiled and soaked oak (p < .05). In conclusion, based on the results of the present study, 10% boiled oak can be substituted corn in diet of laying hens with no unfavourable effect on performance., (© 2020 The Authors Veterinary Medicine and Science Published by John Wiley & Sons Ltd.)

Caffeic acid (CA), a derivative of cinnamic acid, exists in various vegetables and fruits. Natural compounds as beneficial antioxidants play an important role in the research of Alzheimer's disease (AD). In this study, we evaluated the effects of CA against AD using the C. elegans models of AD. We found that CA significantly alleviated Aβ-induced toxicity, increased lifespan, decreased body paralysis, and improved reproductive defects. Also, CA treatment increased resistance to heat and oxidative stress in N2 and CL4176. Moreover, reactive oxygen species (ROS) levels and polyglutamine (polyQ40) aggregate formation were reduced. Furthermore, the mechanistic studies revealed that CA activated transcription factor DAF-16 and its downstream targets SOD-3 and GST-4 to protect against Aβ toxicity. Also, the heat shock proteins hsf-1 and hsp-16.2 mRNA were up-regulated in CL4176. Moreover, CA activated the mRNA expression of lgg-1. Lastly, the KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis of RNA-seq data revealed that there was a significant change in the metabolism-related pathways. Overall, these results suggested an underlying mechanism of action of CA in treating AD at an organism level.

In developing countries, crop deterioration is mainly caused by inappropriate storage conditions that promote insect infestation. Synthetic pesticides are associated with serious adverse effects on humans and the environment. Thus, finding alternative "green" insecticides is a very pressing need. Calotropis procera (Aiton) Dryand (Apocynaceae) growing in Saudi Arabia was selected for this purpose. LC-MS/MS analysis was applied to investigate the metabolic composition of different C. procera extracts. Particularly, C. procera latex and leaves showed a high presence of cardenolides including calactin, uscharidin, 15β-hydroxy-calactin, 16β-hydroxy-calactin, and 12β-hydroxy-calactin. The ovicidal activity of the extracts from different plant organs (flowers, leaves, branches, roots), and of the latex, against Cadra cautella (Walker) (Lepidoptera, Pyralidae) was assessed. Extracts of C. procera roots displayed the most potent activity with 50% of C. cautella eggs not hatching at 10.000 ppm (1%).

The resistance of Haemonchus contortus to synthetic anthelmintics is of increasing concern; and different strategies are being evaluated to improve parasite control. The present study investigated the in vitro effects of combinations of synthetic compounds and monoterpenes. Additionally, the chemical association of the best combinations and their impact on the ultrastructural and biophysical properties of H. contortus eggs was evaluated. We assessed the efficacy of the monoterpenes, carvacrol, thymol, r-carvone, s-carvone, citral, and p-cymene and the anthelmintics, albendazole and levamisole using the egg hatch test (EHT) and the larval migration inhibition test (LMIT), respectively. The minimum effective concentrations of the monoterpenes, according to the EHT (efficacy ranging from 4.4%-11.8%) and LMIT (efficacy ranging from 5.6%-7.4%), were used in combination with different concentrations of synthetic compounds, and the IC 50 and synergism rate (SR) were calculated. Fourier-transform infrared spectroscopy (FTIR) was used to analyze the chemical association between the best combinations as revealed by the in vitro tests (albendazole and levamisole with r-carvone or s-carvone). Atomic force microscopy (AFM) was used to assess the ultrastructural and biophysical properties of H. contortus eggs treated with the albendazole and r-carvone combination. Among the monoterpenes, the highest efficacies were exhibited by carvacrol (IC 50 = 185.9 μg/mL) and thymol (IC 50 = 187.0 μg/mL), according to the EHT, and s-carvone and carvacrol (IC 50 = 1526.0 and 1785.3 μg/mL, respectively), according to the LMIT. According to the EHT, albendazole showed a slight statistically significant synergism in combination with r-carvone (SR = 3.8) and s-carvone (SR = 3.0). According to the LMIT, among the monoterpenes, r-carvone (SR = 1.7) and s-carvone (SR = 1.7) showed an increase in efficacy with levamisole; however, this was not statistically significant. The FTIR spectra of albendazole and levamisole, in association with r-carvone and s-carvone, indicated the presence of chemical interactions between the synthetic and natural molecules, contributing to the possible synergistic effects of these associations. Eggs treated with albendazole and r-carvone showed an increase in roughness and a decrease in height, suggesting that the treatment induced damage to the egg surface and an overflow of its internal contents. Overall, the combination of albendazole with r-carvone and s-carvone was efficacious against H. contortus, demonstrating a chemical association between the compounds; the significant changes in the egg ultrastructure justify this efficacy., (Copyright © 2021 Elsevier B.V. All rights reserved.)

The aim of the present study was to evaluate the effects of dietary supplementation with tomato peels (TP) and rosehip meal (RM) on their phytochemicals transfer to the egg, the antioxidant profile and the markers of lipid peroxidation in stored eggs. The experiment was conducted on 42-week-old TETRA SL laying hens, which were assigned to three dietary treatments with 30 birds each: the Control group (CON) received a corn-soybean diet containing 6% flaxseed meal without any by-product addition. Groups TP and RM received Diet CON with additionally 2% TP and 2% RM, respectively. Average daily feed intake, feed conversion ratio and laying percentage were not significantly changed by the dietary treatments. Compared to the other groups, the inclusion of dietary TP led to a significant increase in egg weight ( p < 0.05) and vitamin E content in eggs ( p < 0.05). The dietary inclusion of RM generated a higher vitamin A content in eggs compared to Groups CON and TP ( p < 0.10). After 40 d of storage, in Groups CON, TP and RM the vitamin E contents of eggs deceased by 10.1%, 14.2% and 13.3%, respectively, whereas the reduction of the lutein and zeaxanthin concentration in eggs was more pronounced (by 24.8%, 21.8% and 25.5%, respectively). For the most primary as well as for secondary oxidation products, storage time had a significant influence ( p < 0.05) on the degradation process of the eggs. After 40 d of storage, the most distinct reduction in the formation rate of secondary oxidation products was observed in Group RM (by more than 30%), while the reduction for Group TP amounted from 15% to 25%, when compared with Group CON. Therefore, the results of the present study showed a significantly positive effect of RM in retarding the lipid peroxidation process during storage.

The probiotic medicinal product TSO (Trichuris suis ova) is administered to patients with active ulcerative colitis in an ongoing clinical phase IIb trial where the typical co-medications are steroids (prednisolone or budesonide) and antibiotics (e.g., phenoxymethylpenicillin). The present pre-clinical study evaluates the effects of these co-medications on the biological activity of TSO in Göttingen Minipigs. This translationally relevant pre-clinical model allows administration of TSO with and without oral steroids or antibiotics in a manner similar to the administration to patients, followed by quantification of the biological activity of TSO. The biological activity of TSO was not affected by oral steroids but was reduced by oral antibiotics. Fecal calprotectin, the common marker of intestinal inflammation in patients with UC, did not differ between groups.

Nine new xanthone-type and anthraquinone-type mycotoxins including austocystins J-N (1-5), 7-chloro versicolorin A (6), 3'-hydroxy-8-O-methyl versicolorin B (7), 8-O-methyl versiconol (8) and 2',3'-dihydroxy versiconol (9), together with 17 known analogues (10-26) were isolated from an extract of the deep-sea-derived fungus Aspergillus puniceus SCSIO z021. Their structures were elucidated by detailed analysis of spectroscopic data, and their absolute configurations were further determined by quantum chemical calculations of ECD spectra or comparison of the experimental ECD spectra. Eleven hydrogenated austocystins were synthesized from 1-2, 10-15 and 17 by catalytic hydrogenation for bioactivities evaluation. Totally, 18 of the all 37 compounds showed strong toxicity against brine shrimps or Vero cell, and the toxicity of 8-O-methyldemethylsterigmatocystin (18) (LC 50  = 0.020 µM) against brine shrimps was higher than those of three positive controls. In addition, 22 of the isolated compounds also exhibited significant inhibitory activity against seven different protein tyrosine phosphatases (PTPs), among them austocystin H (15) and methyl-averantin (24) were the most potent inhibitors with IC 50 values of 0.20-3.0 µM. Their structure-bioactivity relationship was also discussed., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Lead (Pb) is one of the most toxic heavy metal environmental pollutants due to its widespread use of the industry and it is a harmful substance for human and animal health. This study was conducted to investigate the potential protective effects of ellagic acid (EA) on performance, egg quality, antioxidant parameters, and apoptotic pathway proteins in laying quails exposed to Pb toxicity. A total of 168 (15-week old) laying quails (Coturnix coturnix Japonica) were divided into 6 experimental groups (with similar initial average body weight). Birds were fed 1 of 6 diets for 8 weeks: 1 - Control (basal diet), 2 - Pb (basal diet + 100 mg/kg Pb), 3 - EA-300 (basal diet + 300 mg/kg EA), 4 - EA-500 (basal diet + 500 mg/kg EA), 5 - Pb + EA-300 (basal diet + 100 mg/kg Pb + 300 mg/kg EA), 6 - Pb + EA-500 (basal diet + 100 mg/kg Pb + 500 mg/kg EA). The results showed that adding 100 mg/kg of Pb to basal diet was adversely affected the performance parameters and, feed intake and egg production were significantly decreased by Pb supplementation (P < 0.01). However, the EA supplementation to Pb groups improved the performance parameters. Compared with the Pb alone group, in Pb + EA-500 group increased egg production by 8.4%. There were no significant differences in the Haugh unit, albumen index, and yolk index among groups (P > 0.05). Liver and kidney tissues of Pb group malondialdehyde (MDA) level increased (P < 0.001) and, GSH, GSH-Px, and CAT values decreased (P < 0.001) but, EA supplementation alleviated this condition (P < 0.001). The protein levels of caspase-3 and -9 were significantly increased in the Pb group compared to the control group, whereas EA supplementation alleviated the Pb-induced apoptosis by decreasing caspase-3 and -9 levels in the liver tissue (p < 0.001). In laying quails exposed to Pb toxicity, EA supplementation improves the performance parameters, enhances the antioxidant defense system, and suppresses apoptosis via regulates the expression of caspase-3 and -9. Thus, it was concluded that EA (especially 500 mg/kg) can ameliorate the toxic effects of Pb exposure in quails., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

Ethnopharmacological Relevance: Propolis has been used in folk medicine for thousands of years and, in the past few decades, it has attracted renewed interest. Although propolis has been traditionally used in many communities worldwide against parasitic diseases, its effect against Schistosoma mansoni infection remains unclear., Aim of the Study: To demonstrate the effects of Brazilian red propolis on Schistosoma mansoni ex vivo and in an animal model of schistosomiasis., Materials and Methods: In vitro, we monitored phenotypic and tegumental changes as well as the effects of the crude extract of propolis on pairing and egg production. In a mouse infected with either immature (early infection) or adult (chronic infection) worms, propolis was administered by oral gavage and we studied the influence of this natural product on worm burden and egg production., Results: Propolis 25 μg/mL reduced motility and caused 100% mortality of adult parasites ex vivo. Further analysis revealed a pronounced reduction in oviposition after exposure to propolis at sub-lethal concentrations. In addition, scanning electron microscopy showed morphological alterations in the tegument of schistosomes. In the animal model, propolis markedly reduced worm burden and egg production in both early and chronic S. mansoni infection when compared to untreated control animals., Conclusions: The efficacy of Brazilian red propolis in both in vitro and in vivo studies suggests its potential anthelmintic properties against S. mansoni infection., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This experiment examines the effects of bamboo charcoal (BC) powder, bamboo vinegar (BV), and their combination (BCV) in the diet of laying hens on performance, egg quality, relative organ weights, and intestinal bacterial populations. A total of 320 laying hens (36 weeks of age) were divided into 4 treatment groups, with 10 replicates per treatment and 8 hens per replicate. They were fed on a control diet, the control diet supplemented with 0.8% BC, the control diet supplemented with 0.4% BV, or the control diet supplemented with a combination of BC (0.8%) and BV (0.4%) from 36 to 51 weeks of age. Egg production increased in the hens fed the BV and BCV diets during 48 to 51 weeks of age (P < 0.05). Damaged egg rate decreased in the hens fed the BV and BCV diets for the whole experiment (P < 0.05). Shell thickness was highest in the BCV-fed group at week 43, and shell strength was higher in the BV-fed group at week 51 (P < 0.05). Supplementation of BC or BCV in the diet resulted in a decreased abdominal fat pad (P < 0.05). In the ileal content, the population of Salmonella spp. decreased in the BV and BCV groups and the population of Lactobacillus spp. increased in the BV group (P < 0.05). The present results indicate that feeding BV or BCV alleviates damaged egg rate and decreases intestinal pathogenic bacteria, while feeding BC benefits by reducing abdominal fat. These results suggest that the effect of BCV seems to be induced by the synergistic effect of BC and BV, and that the BCV contributes to the effective use of bamboo on the laying hen's production.

A multitude of factors affect the assemblies of complex microbial communities associated with animal hosts, with implications for community flexibility, resilience, and long-term stability; however, their relative effects have rarely been deduced. Here, we use a tractable lab model to quantify the relative and combined effects of parental transmission (egg case microbiome present/reduced), gut inocula (cockroach versus termite gut provisioned), and varying diets (matched or unmatched with gut inoculum source) on gut microbiota structure of hatchlings of the omnivorous cockroach Shelfordella lateralis using 16S rRNA gene (rDNA) amplicon sequencing. We show that the presence of a preexisting bacterial community via vertical transmission of microbes on egg cases reduces subsequent microbial invasion, suggesting priority effects that allow initial colonizers to take a strong hold and which stabilize the microbiome. However, subsequent inoculation sources more strongly affect ultimate community composition and their ecological networks, with distinct host-taxon-of-origin effects on which bacteria establish. While this is so, communities respond flexibly to specific diets in ways that consequently impact predicted community functions. In conclusion, our findings suggest that inoculations drive communities toward different stable states depending on colonization and extinction events, through ecological host-microbe relations and interactions with other gut bacteria, while diet in parallel shapes the functional capabilities of these microbiomes. These effects may lead to consistent microbial communities that maximize the extended phenotype that the microbiota provides the host, particularly if microbes spend most of their lives in host-associated environments. IMPORTANCE When host fitness is dependent on gut microbiota, microbial community flexibility and reproducibility enhance host fitness by allowing fine-tuned environmental tracking and sufficient stability for host traits to evolve. Our findings lend support to the importance of vertically transmitted early-life microbiota as stabilizers, through interactions with potential colonizers, which may contribute to ensuring that the microbiota aligns within host fitness-enhancing parameters. Subsequent colonizations are driven by microbial composition of the sources available, and we confirm that host-taxon-of-origin affects stable subsequent communities, while communities at the same time retain sufficient flexibility to shift in response to available diets. Microbiome structure is thus the result of the relative impact and combined effects of inocula and fluctuations driven by environment-specific microbial sources and digestive needs. These affect short-term community structure on an ecological time scale but could ultimately shape host species specificities in microbiomes across evolutionary time, if environmental conditions prevail., (Copyright © 2021 Renelies-Hamilton et al.)

Diapause represents a major developmental switch in insects and is a seasonal adaptation that evolved as a specific subtype of dormancy in most insect species to ensure survival under unfavorable environmental conditions and synchronize populations. However, the hierarchical relationship of the molecular mechanisms involved in the perception of environmental signals to integration in morphological, physiological, behavioral, and reproductive responses remains unclear. In the bivoltine strain of the silkworm Bombyx mori , embryonic diapause is induced transgenerationally as a maternal effect. Progeny diapause is determined by the environmental temperature during embryonic development of the mother. Here, we show that the hierarchical pathway consists of a γ-aminobutyric acid (GABA)ergic and corazonin signaling system modulating progeny diapause induction via diapause hormone release, which may be finely tuned by the temperature-dependent expression of plasma membrane GABA transporter. Furthermore, this signaling pathway possesses similar features to the gonadotropin-releasing hormone (GnRH) signaling system for seasonal reproductive plasticity in vertebrates., Competing Interests: The authors declare no competing interest., (Copyright © 2021 the Author(s). Published by PNAS.)

This study was conducted to determine the effects of supplemental dietary benzoic acid on production performance, egg quality, intestinal morphology, and intestinal microbiota of laying hens. A total of seven hundred twenty 45-wk-old Lohman pink-shell laying hens were randomly allocated to 3 dietary treatments: control (CON), diet supplemented with 1,000 mg/kg benzoic acid (BA1), and 2,000 mg/kg benzoic acid (BA2). Each treatment included 10 replicates of 24 hens; laying hens were monitored for 16 wk. Overall, the results indicate that benzoic acid supplementation had no effect on laying rate, feed intake, feed conversion ratio, and breaking rate; however, a decrease in egg weight (P < 0.01) was observed in the BA2 group. Albumen height and Haugh unit (HU) were also linearly increased in the BA1 and BA2 groups (linear effect, P < 0.05). An increase in duodenum villus height (V) (quadratic effect, P = 0.041) and crypt depth (C) (linear effect, P = 0.012) was observed in the BA2 group, whereas an increased jejunum C and decreased V/C (quadratic effect, P < 0.05) in the BA1 group. Moreover, an increase in ileum V and C (quadratic effect, P < 0.05) was observed in the BA1 group. Microbial richness and diversity were reduced in the BA2 group (P < 0.01). An increase in the abundance of Clostridia (class), Clostridiales (order), Ruminococcaceae (family), and Lachnospiraceae (family) was noted in the BA1 group, whereas an enrichment of Bacteroides caecicola (species) was observed in the BA2 group. The HU positively correlated with genus Sphaerochaeta and Enorma (r = 0.56, 0.56; P < 0.05) but negatively correlated with Romboutsia, Subdoligranulum, Helicobacter, and Mucispirillum (r = -0.58, -0.49, -0.48; -0.70; P < 0.05). In conclusion, dietary supplementation with benzoic acid had no effect on production performance, but it significantly improved egg quality. In addition, 1,000 mg/kg benzoic acid positively modulated intestinal health by improving intestinal morphology and enriching microbial composition., (Copyright © 2020. Published by Elsevier Inc.)

Background: Organic products of animals are getting more accepted by consumers. Using herbal additives may lead to more health animal products. In this research it is hypothesized that Lavandula angustifolia and/or Mentha spicata essential oils would be helpful to enhance production performance in laying hens., Objectives: This experiment was conducted to evaluate the effects of Lavandula angustifolia and Mentha spicata essential oils on performance, egg traits and blood variables in laying hens., Methods: 144 Lohmann LSL-Lite laying hens from 42 until 56 weeks of age were used in a completely randomized design in four treatments and six replicates (six birds per replicate). The treatments consisted of: (a) control group (basal diet), (b) basal diet supplemented with 250 mg/kg diet lavender essential oil (LEO), (c) basal diet supplemented with 250 mg/kg diet mint essential oil (MEO), and (d) basal diet supplemented with both LEO and MEO., Results: Using LEO and/or MEO did not affect body weight changes, feed intake, egg weight, egg index, yolk index, Haugh unit, egg shell weight and egg shell thickness. Feeding LEO, individually or in combination with MEO, did not affect FCR compared with the control group (p < .05), however, feeding MEO individually increased feed conversation ratio (FCR) compared to LEO and the control group during 42-56 weeks (p < .05), as well as decreasing egg mass compared to LEO (p < .05). Feeding LEO increased egg production compared to MEO and combination of MEO and LEO (p < .05)., Conclusions: In conclusion, dietary supplemental MEO (250 mg/kg) may increase FCR, and LEO (250 mg/kg) is more effective than MEO (250 mg/kg) for egg production and egg mass purposes; besides MEO (250 mg/kg) negatively affected FCR compared with the control group. In addition, no specific beneficial effect of dietary supplemental MEO and/or LEO on the other measured variables was detected., (© 2020 The Authors. Veterinary Medicine and Science published by John Wiley & Sons Ltd.)