Effect of dietary supplementation of wild leek (Allium tricoccum) and garlic (Allium sativum) leaves on production, egg quality, serum lipid profile, intestinal morphology and nutrient digestibility of laying quails.

In this study, the effects of adding dried wild leek and garlic leaves to the diet of laying quails on egg quality, production performance, intestinal histomorphology, nutrient digestibility, and serum biochemical traits were evaluated. 168- 30 weeks aged- laying quails were randomly arranged into 7 treatments (control group and three different levels (0.5%, 1%, and 1.5%) of wild leek or garlic leaves), 6 replicates, and 4 quails per pen. According to the study, the highest hen-day production was achieved by adding 0.5% garlic leaves, and the egg weight was higher with 1% garlic leaves and 0.5% wild leek compared to the control group (P < 0.05). In addition, adding garlic leaves decreased the average daily feed intake and increased high-density lipoprotein concentration. However, adding both herbs did not influence yolk height, albumin height, shell thickness, and Haugh unit (P > 0.05). All supplemented treatments significantly increased the formerly Roche Yolk Color Fan (DSM) index compared to the control. Quails fed with 1% garlic leaves declined serum cholesterol, very low-density lipoprotein, low-density lipoprotein, and uric acid. Moreover, three levels of garlic leaves decreased triglyceride concentration. Adding 0.5% and 1% garlic leaves to the diet could increase the jejunal, and ileal villus height-to-crypt depth ratio. Furthermore, 1% and 1.5% garlic leaves supplementation elevated organic matter, ash, and crude protein digestibility. In conclusion, supplementing 1% garlic leaves to the quail's diet could improve egg weight, intestinal morphology, and nutrient digestibility, improving lipid profiles and reducing uric acid concentration in the serum. However, both plants were rich pigment sources, increasing the DSM index.

Toxic effects of freshwater grouper (Acrossocheilus fasciatus) eggs on poultry: Morphological and transcriptomic insights into hepatic toxicity.

Fish egg poisoning is a serious and neglected public menace that kills hundreds of people and numerous poultry each year. Freshwater groupers (Acrossocheilus fasciatus) are common food fish in the southeastern regions of China. Their toxic eggs are regarded as a significant public health concern. The molecular mechanisms of egg-toxin toxicity in freshwater grouper to poisoned organisms are elusive. In this study, black-boned chicks were exposed to toxic eggs from freshwater grouper at a lethal dose. The hepatic morphology of the intoxicated chick was assessed. An analysis of the liver gene expression profile was conducted by comparing samples exposed to toxic eggs with control samples using RNA-Seq. The result revealed that an increase in vacuolation and congestion was observed in chicks with toxic eggs exposure. The transcriptome analysis revealed 5421 genes with differential expression, comprising 2810 up-regulated and 2611 down-regulated genes. The genes were primarily linked to energy metabolism, cell apoptosis, cell adhesion, exogenous microbial infection, and cell junction. The most strongly upregulated genes were cholecystokinin (CCK), cholecystokinin A receptor (CCKAR), and unc-80 homolog, NALCN activator (UNC80), and the most downregulated genes were glycine amidinotransferase (GATM), fatty acid desaturase 2 (FADS2), and hexokinase 2 (HKDC1). GO term with the highest enrichment of DEGs is nucleosome assembly. According to KEGG pathways, the three most significant metabolic pathways in the liver are DNA replication, retinol metabolism, and steroid biosynthesis. The results could be crucial for comprehending the negative biological impacts of egg-toxin and its toxic mechanisms. The outcome could provide potential biomarkers of egg-toxin exposure in hepatic, which might be useful for manufacturing an antidote to egg-toxin and providing valuable insights for ecotoxicity studies.

Exposure of female lobsters to sediments spiked with emamectin benzoate: Effects on eggs and larvae early development.

The salmon aquaculture industry is an important economic activity established on both the west and east coast of Canada. To control sea lice infestations, in-feed products like emamectin benzoate (EMB) are widely used. Due to its low solubility and persistence EMB can accumulate in marine sediments and be potentially bioavailable to non-target organisms from months to years. The American lobster (Homarus americanus) is a key species in the Northwest Atlantic with high economic and ecological value. It may be exposed to therapeutants considering lobster habitats overlap with aquaculture locations requiring a better understanding of the potential impact of these therapeutants through varied pathways of exposure. In this study, we investigated the exposure of gravid female lobsters to EMB spiked sediment to mimic the likely presence of these females at aquaculture sites for a 10-day period. We completed testing by assessing EMB effects on adult molting and quality, embryo hatching rates, and larval offspring quality and larval molting. Our results show that a single, 10-day exposure of ovigerous females to EMB concentrations higher than environmentally relevant values did not affect females or their offspring.

Ovicidal activity and cytotoxicity of ethanolic extract of turmeric (Curcuma longa) and green tea (Camellia sinensis) to treat digestive parasite of sheep.

Background: The Trichuris eggs are collected from naturally infected sheep. Natural antihelmintics such as herbal medicines are needed as an alternative, such as natural compounds from endemic plants. Aim: This present study aims to evaluate the ovicidal activity and cytotoxicity effects of ethanolic extract of Curcuma longa (EECL) and Camelia sinensis (EECS) as a biological anthelmintic against the egg of Trichuris sp. Methods: The Trichuris eggs are collected from naturally infected sheep. CMC-Na solution 1% was used as a control. The treatments were 0.12% EECL; 0.24% EECL; 0.15% EECS; 0.30% EECS; a combination of 0.12% EECL and 0.30% EECS; a combination of 0.24% EECL; and 0.15% EECS. Ovicidal activity testing by microscopic examination of eggs treated using different concentrations of EECL extract, EECS, and a combination of them. They were exposed for various times (7, 14, 21, and 28 days) and incubated at room temperature. Results: The study shows that a combination of C. longa extract and tea extract exhibits good ovicidal anthelmintic activity against Trichuris sp. in sheep. Cytotoxicity examination using the 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyl tetrazolium bromide (MTT) test. Based on MTT data processed using regression analysis, the obtained LC50 from the administration of EECL, EECS, and a combination of both in a ratio of 1:1, 2:2, 1:2, and 2:1. The combination of EECL extract and EECS with the highest concentration produced cell viability of 28.46%, 17.25%, 56.01%, and 46.47%, respectively. Conclusion: It can be concluded that the most cytotoxic ingredient is found in the combination of EECL and EECS (2:2) at 17.25% and the safest is in the ratio (1:2) at 56.01%.

Exploring the effects of varying levels of selenium-chitosan on production performance, egg quality, and immune health in laying japanese quail.

The purpose of this research was to see how different levels of Se-chitosan, a novel organic source of Se, affected the production performance, egg quality, egg Se concentration, microbial population, immunological response, antioxidant status, and yolk fatty acid profile of laying Japanese quail. This experiment used a totally randomized design, with 5 treatments, 6 repeats, and 10 birds in each repetition. The dietary treatment groups were as follows: no Se supplementation (control group), 0.2 mg/kg Na-selenite supplementation, and 0.2, 0.4, and 0.6 mg/kg Se-chitosan supplementation. The feed conversion ratio (FCR) improved linearly in quails fed different levels of Se-chitosan compared to the control group (P < 0.05). Furthermore, Se-chitosan at concentrations of 0.2 and 0.4 mg/kg demonstrated both linear and quadratic increases in albumen height, Haugh unit, and yolk color in fresh eggs compared to the control group. Additionally, Se-chitosan contributed to enhanced shell thickness and strength, along with an increased Se concentration in the yolk. Se-chitosan supplementation at different levels linearly and quadratically reduced coliforms (COL) while increasing lactic acid bacteria (LAB)/coliform ratios (P < 0.05). Se-chitosan supplementation linearly and quadratically increased the total antibody response to sheep red blood cells (SRBC) and IgG titers (P < 0.05). It also linearly decreased the level of malondialdehyde in fresh and stored egg yolks and increased the activity of antioxidant enzymes catalase and glutathione peroxidase linearly, and superoxide dismutase (SOD) both linearly and quadratically in quail blood serum (P < 0.05). Additionally, supplementation of Se-chitosan at levels of 0.2 and 0.6 mg/kg linearly decreased the ∑ n-6 PUFA/∑ n-3 PUFA ratio in the yolk compared to the control group (P < 0.05). It can be concluded that incorporating Se-chitosan as a novel organic source of Se in the diet of laying quails can enhance production performance, egg quality, egg Se concentration, yolk lipid oxidation, microbial population, immune response, antioxidant enzyme activity, and yolk fatty acid profile.

Effects of immersing Japanese quail eggs in various doses of riboflavin on reproductive, growth performance traits, blood indices and economics.

This investigation aimed to evaluate the impact of immersion (IM) riboflavin treatment on the hatchability, production efficiency, and carcass characteristics of Japanese quail eggs. A total of 260 eggs of Japanese quail birds were used for hatching and were randomly divided into 4 treatments with 5 replicates (13 eggs/replicate) in a fully randomized design. Hatching eggs were immersed in riboflavin for 2 min before incubation. The experiment treatments were designed as follows: G1 control group with no treatment, G2 treated with 3 g/L vit. B2 (IM), G3 treated with 4 g/L vit. B2 (IM) and G4 were treated with 5 g/L vit. B2 (IM). After hatching, 128 Japanese quail chicks, aged 7 d, were randomly grouped into 4 treatment groups, with 32 birds in each group. When quails were given vitamin B2 via immersion, they demonstrated significant enhancements in live body weight, body weight gain, feed consumption, and feed conversion ratio at different stages compared to the control group. Compared to control and other groups, the carcass parameters of Japanese quails given a 4 g/L immersion solution showed a significant improvement (P < 0.05). Hatchability and fertility (%) were considerably raised by Vit.B2 treatments of 3, 4, and 5g; the group immersed in 5 g/L had the highest percentages compared to the other groups. Furthermore, treated chickens with all concentrations of vitamin B2 had significantly higher blood indices than the controls. During the exploratory phase (1-6 wk) of age, the highest returns were reported in G4 treated with 5g/L vit. B2 (IM). Treating Japanese quail eggs with different dosages of vitamin B2 by immersion may be recommended to improve their productive and reproductive performance, blood indices, carcass traits, and economic efficiency.

Impact of in ovo feeding of grape pomace extract on the growth performance, antioxidant status, and immune response of hatched broilers.

Delivering natural antioxidants via in ovo feeding holds promise for enhancing the antioxidant status and performance of chickens. Therefore, The objective of this study was to evaluate the impacts of in ovo feeding during early embryonic development using grape pomace extract as a natural antioxidant on hatchability, productive performance, immune response, and antioxidant status in broilers. A total of 900 fertile broiler eggs from the Arbor Acres strain were utilized. Each egg was individually weighed, with egg weights ranging from 61.88 ± 3 g. On the 17.5th d of incubation (DOI), the fertile eggs were divided into 6 groups. The first treatment group was untreated and designated as the control (C). The second group was the sham group (Sh), receiving a simulated injection. The third group, designated as the vehicle group (V), was injected with 100 µl of dimethyl sulfoxide (DMSO). The fourth group received an injection of 100 µL of grape pomace dissolved in DMSO at a concentration of 2 mg (T2). Similarly, the fifth and sixth groups were injected with 100 µL of grape pomace dissolved in DMSO at concentrations of 4 mg and 6 mg, (T4), (T6) respectively. Subsequently, all groups were raised under uniform conditions in terms of management, environment, and nutrition till 5 wk of age. The grape pomace extract (GPE), obtained is rich in total phenolic content (16.07 mg/g), total flavonoid content (7.42 mg/g), and total anthocyanin (8.37 mg/g). Grape pomace extract has exhibited significant antioxidant properties as evidenced by its effectiveness in DPPH scavenging and reducing power assays. Significant improvements in body weight at hatch were observed with in ovo feeding of grape pomace extract, particularly at the 4 mg level, surpassing the effectiveness of the 2 mg and 6 mg grape pomace levels, and this enhancement in body weight continued until the age of 5 wk. GPE injection also led to a significant reduction in cholesterol levels, with the lowest levels recorded for the T4 group. Plasma total Antioxidant Capacity (TAC) levels were significantly elevated in groups treated with T4, T6, and T2 compared to the control group. Conversely, the control group showed a significant increase (P < 0.01) in plasma malondialdehyde (MDA) levels. The immune response of hatched chicks from grape pomace extract-injected groups, especially the T4 group, exhibited improvement through increased IgM and IgG. These findings demonstrate that in ovo feeding of GPE, particularly at a dosage of 4 mg, enhances growth performance, immune response, and antioxidant status in hatched chicks. Thus, administering natural antioxidants, such as grape pomace extract, to developing broiler embryos via in ovo feeding could serve as a valuable strategy for enhancing the subsequent post-hatch productive performance, as well as bolstering the antioxidant and immunological status of broiler chicks.

Revolutionizing poultry hygiene: advanced electrostatic and cold fog disinfection strategies combat Mycoplasma gallisepticum in hatching eggs.

The incidence of chronic respiratory disease (CRD) due to Mycoplasma gallisepticum (MG) contamination in hatching eggs poses a serious threat to poultry health and hatchability. Implementing effective sanitization methods while safeguarding the hatching potential of embryos is crucial. This study aimed to explore novel techniques for sanitizing hatching-fertile eggs to prevent and manage MG-associated CRD. The primary objective was to assess the efficacy of acidic electrochemically stimulated water (ECS), focusing on MG disinfection. Additionally, the study investigated 2 application methods, 1) electrostatic disinfection (ED) and 2) cold fog (CF) disinfection, to evaluate their bactericidal effects against MG-contaminated eggs. Deliberately infected MG strains were used for the experimental design, which compared the disinfection efficacy of ECS with its acidic properties. The comparison involved ED, which applies an electrostatic charge to water particles, and CF disinfection, a cold mist technique. Both methods aimed to target MG without compromising egg-hatching potential. The results indicated a significant (p < 0.05) reduction in colony-forming units per milliliter (CFU/mL). However, both application methods demonstrated distinct bactericidal effects. Eggs treated with electrostatic disinfection showed a significant (p < 0.001) reduction in embryonic mortality during incubation (10%) compared to control untreated eggs (18%). Similarly, the CF method exhibited a significant (p < 0.001) decrease in embryonic mortality (13%). The ECS potential in reducing embryonic mortality within the pH range of 2.5 to 6.5 was noted. Both the ED and CF methods show promise for preventing MG-induced hatchery infection while maintaining egg-hatching potential. This study presents innovative techniques to control MG in hatching eggs, contributing to improved poultry health and reduced CRD incidence.

The potential of in ovo-fed amino acids to alleviate the effects of heat stress on broiler chickens: effect on performance, body temperature, and oxidative status during the finisher phase.

The aim of the current study was to investigate the potential of in ovo-fed amino acids (AA) to reduce the effects of heat stress on finishing broiler chickens. To achieve this, a total of 1,400 fertile hatching eggs were randomly distributed into 5 groups (n = 280/group) and injected with one of the following in ovo treatments on embryonic day 18: 52 µL of sterile diluent/egg (CTRL), CTRL + 1.0 mg of L-Leucine (T1), CTRL + 0.45 mg of leucine + 1.15 mg of methionine (T2), CTRL + 3.0 mg of methionine + 2.0 mg of cysteine (T3), and CTRL + 0.40 mg of leucine + 1.60 mg of methionine + 1.60 mg of cysteine (T4). After hatch, chicks were allocated according to a complete randomized block design comprising 2 thermal conditions: thermoneutral (24°C, 45% RH) and heat stress (34°C, 55-60% RH) with 5 pens/group/condition. The cyclical heat stress regimen (10 h/d) was then applied from d 29 to d 34. Compared to the CTRL group, T3 and T4 exhibited a higher BW during the starter phase (P < 0.001). T4 also had a lower feed conversion ratio (FCR) than CTRL during this same phase (P = 0.03). During the grower phase, males of all treatment groups consistently exhibited higher BW compared to the CTRL group, which was not observed among female birds (PSex × TRT = 0.005). During the finisher phase, the in ovo treatment effect on performance was not significant. However, heat-stressed birds from treatment group T3 and T4 exhibited lower facial temperatures (Pday × TRT < 0.001) as well as lower plasma (Pcondition x TRT = 0.039) and liver (Pcondition x TRT < 0.001) malonaldehyde concentrations compared to the CTRL group. In conclusion, in ovo-fed AA have the potential to modulate the effects of heat stress on finishing broiler chickens by limiting its detrimental consequences, including increased body temperature and oxidative damage.

Compatibility of synthetic and biological pesticides with a biocontrol agent Phytoseiulus longipes (Acari: Phytoseiidae).

Phytoseiulus longipes is a predatory mite of Tetranychus evansi, which is an invasive pest in Africa and elsewhere. The introduction of this predator in Africa has considerable potential, but little is known about the compatibility of P. longipes with commonly used pesticides. Here, we examined lethal and sublethal effects of two pyrethroids (cypermethrin and deltamethrin), two organophosphates (dimethoate and chlorpyrifos), one nicotinoid (imidacloprid), two acaricides (propargite and abamectin), two naturally derived pesticides (oxymatrine and azadirachtin), and one entomopathogenic fungal-based formulation (Hirsutella thompsonii) on P. longipes eggs and adults. The pesticides were sprayed at their maximum recommended concentrations. Topical exposures to azadirachtin, imidacloprid, propargite, abamectin, oxymatrine, and H. thompsonii significantly reduced the net reproductive rate (R0), intrinsic rate of increase (r) and finite rate of increase (λ)of P. longipes. Pesticide lethal and sublethal effects on the predator were summarized in a reduction coefficient (Ex) for the classification based on IOBC toxicity categories. Results revealed that Azadirachtin and H. thompsonii were slightly harmful effects to adults. Imidacloprid, propargite, abamectin, and oxymatrine were moderately harmful to both eggs and adults. Residual persistence bioassays revealed that 4-day-old residue of azadirachtin had no harmful effect on the predator. Abamectin, oxymatrine, and H. thompsonii became harmless to it 10 days post-spraying, and propargite and imidacloprid were considered harmless after 20 days. Cypermethrin, deltamethrin, dimethoate, and chlorpyrifos were highly harmful to both eggs and adults, persistence remaining high even after 31 days of application. These findings provide valuable insights into decision-making when considering P. longipes for use in IPM programs.

Ozone and peroxone disinfection of Toxocara canis eggs in water.

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.

Assessment of the effects of 2,4-D-dichlorophenoxyacetic acid-based herbicide exposure in eggs and embryos of golden apple snails using OCT.

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.

Bioactivities of essential oil extracted from Elsholtzia densa Benth. And its main components against Tribolium castaneum eggs and pupae.

This study aimed to develop a relatively natural and safe botanical insecticide for controlling the storage pest Tribolium castaneum in the egg and pupal stages. It examined how Elsholtzia densa Benth. essential oil (EO) and its primary components, ß-caryophyllene and limonene, affected T. castaneum eggs and pupae through contact and fumigation. Among th, the contact activities of ß-caryophyllene against T. castaneum eggs and pupae are LD50 (median lethal dose, 50%) = 0.156 mg/cm2 and ED50 (median effective dose, 50%) = 16.35 mg/pupa respectively. The study also investigated the effect of ß-caryophyllene and limonene on T. castaneum eggs and pupae through synergistic contact and fumigation. When the mixing ratio of ß-caryophyllene and limonene was 7:1, the LD50 value of contact activity against T. castaneum eggs was reduced to 0.100 mg/cm2, displaying an obvious synergistic effect. Experiments were conducted to investigate the antitoxic effect of ß-caryophyllene on T. castaneum eggs and pupae, as well as its effects on the enzymatic activity of acetylcholinesterase, succinate dehydrogenase, glutathione S-transferase and carboxylesterase in T. castaneum pupae. Finally, the molecular docking techniques were employed to confirm the aforementioned effects on enzyme function. The findings of this study might help improve storage pest control with T. castaneum and create eco-friendly insecticides using E. densa EO, ß-caryophyllene, and limonene.

Peracetic acid treatment of squid eggs infected with parasitic copepod (Ikanecator primus gen. et sp. nov.).

Having been successfully bred in semi-intensive and intensive aquaculture systems, oval squids of the Sepioteuthis lessoniana species complex are emerging as promising candidates for research and industry. Nevertheless, information about pathogens and diseases that may affect squid aquaculture remains sparse. In this study, we identify new parasitic copepod species that causes squid mortality and decreases squid hatching rates, and we also offer a solution to eliminate the pathogen during incubation of squid eggs. The newly discovered copepod Ikanecator primus gen. et sp. nov. was identified on oval squid eggs for the first time using both morphological and molecular diagnostic markers. In the genomes of the copepod and associated microbiome, we identified multiple genes for enzymes involved in cephalopod eggshell degradation in genomes of the copepod and associated microbiome. Furthermore, we conducted experiments to assess efficacy of peracetic acid in inhibiting the I. primus gen. et sp. nov. both in vitro and in vivo using immersion treatment. We established that a 2-min exposure to a concentration of 250 µl/L of peracetic acid containing product (PAA-product; 35 mg/L PAA and 15 mg/L H2O2) inhibited the development of nauplii in vitro. All parasites exposed to a concentration of 500 µl/L of PAA-product (70 mg/L PAA and 30 mg/L H2O2) were eliminated within two minutes. On top of this, the immersion treatment with 500 µl/L of PAA-product (70 mg/L PAA and 30 mg/L H2O2) improved survival of squid embryos and increased size of squid hatchlings compared with control and the immersion treatment with 125 µl/L of PAA-product (17.5 mg/L PAA and 7.5 mg/L H2O2) and the immersion treatment with 250 µl/L of PAA-product (35 mg/L PAA and 15 mg/L H2O2). These findings suggest that PAA holds a great potential as inhibitor and controller of parasitic copepod infections and for overall health management in cephalopod culture.

A taste of a toxin paradise: Xenorhabdus and Photorhabdus bacterial secondary metabolites against Aedes aegypti larvae and eggs.

Aedes-transmitted arboviral infections such as Dengue, Yellow Fever, Zika and Chikungunya are increasing public health problems. Xenorhabdus and Photorhabdus bacteria are promising sources of effective compounds with important biological activities. This study investigated the effects of cell-free supernatants of X. szentirmaii, X. cabanillasii and P. kayaii against Ae. aegypti eggs and larvae and identified the bioactive larvicidal compound in X. szentirmaii using The EasyPACId method. Among the three tested bacterial species, X. cabanillasii exhibited the highest (96%) egg hatching inhibition and larvicidal activity (100% mortality), whereas P. kayaii was the least effective species in our study. EasyPACId method revealed that bioactive larvicidal compound in the bacterial supernatant was fabclavine. Fabclavines obtained from promoter exchange mutants of different bacterial species such as X. cabanillasii, X. budapestensis, X. indica, X. szentirmaii, X. hominckii and X. stockiae were effective against mosquito larvae. Results show that these bacterial metabolites have potential to be used in integrated pest management (IPM) programmes of mosquitoes.

Effect of cattle and horse feces storage methods on Nematode egg viability and sensitivity for egg hatch test.

The aim of the present study was to validate methods of stool sample conservation for the egg hatch test (EHT). This study involved the use of a bovine naturally infected predominantly by Cooperia spp. and one equine naturally infected predominantly by cyathostomins characterized as susceptible to benzimidazoles in the EHT. Fecal samples were submitted to three treatments: aerobic methods (anaerobic storage in plastic bottles, anaerobic storage in vacuum-sealed bags or aerobic storage in plastic bags), under two temperature conditions (room temperature and refrigeration) analyzed at four different assessment times (48, 72, 96 and 120 h). As the standard test, an assay was also performed within 3 h. The tests were performed in triplicate for each drug concentration and with three experimental repetitions at one-week intervals. Two criteria were used for the storage methods: hatchability in the negative control group and sensitivity of the eggs to thiabendazole, comparing the EC50 and 95% confidence interval for each treatment to those of the standard test and the other repetitions. Bovine samples can be stored for up to 96 h and refrigerated vacuum storage can be used, ensuring hatchability of the negative control and sensitivity of the eggs to thiabendazole. For equine samples, no forms of storage were indicated due to the variation among the repetitions and the reduction in the sensitivity of the eggs to thiabendazole, which could result in a false positive detection of resistance.

In vitro ovicidal and larvicidal activity of a hydroalcoholic extract and its fractions from Cyrtocarpa procera fruits on Haemonchus contortus.

This study describes the in vitro anthelmintic effect of a hydroalcoholic extract (HA-E) and its fractions from Cyrtocarpa procera fruits against Haemonchus contortus eggs and infective larvae. The HA-E was subjected to bipartition using ethyl acetate, which resulted in an aqueous fraction (Aq-F) and an organic fraction (EtOAc-F). The HA-E and both fractions were tested using the egg hatching inhibition assay (EHIA) and the larval mortality test (LMT). Fractionation of the EtOAc-F was achieved using different chromatographic processes, i.e., open glass column and HPLC analysis. Fractionation of the EtOAc-F gave 18 subfractions (C1R1-C1R18), and those that showed the highest yields (C1R15, C1R16, C1R17 and C1R18) were subjected to anthelmintic assays. The HA-E and the EtOAc-F displayed 100% egg hatching inhibition at 3 and 1 mg/mL, respectively, whereas Aq-F exhibited 92.57% EHI at 3 mg/mL. All subfractions tested showed ovicidal effect. Regarding the larval mortality test, HA-E and EtOAc-F exhibited a larvicidal effect higher than 50% at 50 and 30 mg/mL, respectively. The subfractions that showed the highest larval mortality against H. contortus were C1R15 and C1R17, with larval mortalities of 53.57% and 60.23% at 10 mg/mL, respectively. Chemical analysis of these bioactive subfractions (C1R15 and C1R17) revealed the presence of gallic acid, protocatechuic acid, and ellagic acid. This study shows evidence about the ovicidal and larvicidal properties of C. procera fruits that could make these plant products to be considered as a natural potential anthelmintic agents for controlling haemonchosis in goats and sheep.

Response of laying hens fed diet supplemented with a mixture of olive, laurel, and rosemary leaf powders: Metabolic profile, oxidative status, intestinal histomorphology, and egg quality.

This study aimed to evaluate the effects of a mixture of olive, laurel, and rosemary leaf powders, on the oxidative state, biochemical, immune, intestinal morphophysiological parameters, and egg quality of laying hens. One hundred Lohmann Brown hens (28 weeks old) were equally assigned to two groups (n. 50) corresponding to a basal control diet (CON) or the diet supplemented with 6 g/kg feed of leaf powder mixture (LPM) containing olive, laurel, and rosemary leaves (1:1:1), for 60 days. Oxidative status, biochemical indices, immune response, cecal short chain fatty acids (SCFAs), intestinal morphological characteristics, and some egg traits were evaluated at the end of the experiment. The results indicated that LPM improved (P < 0.05) the oxidative status (TOS, ROMs), the immune system (IL-6, IL-1ß, and TNF-α), the total protein and HDL cholesterol content, whereas it decreased (P < 0.05) total cholesterol and LDL cholesterol. Aspartate aminotransferase (AST), alkaline phosphatase (ALP), and alanine aminotransferase were significantly (P < 0.05) lower in the LPM than in the CON group. A significant increase (P < 0.05) in SCFA content in the caecum, as well as in villi height and crypt depth in both duodenum and ileum of LPM-treated hens, was observed. Egg quality parameters were not influenced (P > 0.05) by LPM. These findings indicate that LPM can be considered a candidate as an antioxidant ingredient for functional food in laying hens.

Synergistic Interaction Effect of Artemisia cina n-hexane Extract and Tagetes lucida Ethyl Acetate Extract on Haemonchus Contortus.

PURPOSE: We analysed the possible synergistic activity among active extracts from Artemisia cina and Tagetes lucida combinations on Haemonchus contortus, a nematode parasitising sheep. METHODS: The work was carried out in vitro on eggs and infective larvae (L3) of H. contortus. The results were analysed with SAS 9.1, applying the ANOVA and Tukey test, and the lethal concentration (LC) values LC50 and LC90 were determined with regression analysis, employing Proc Probit of SAS 9.1. Additionally, the lethal concentration (LC) was calculated with LC50 and LC90 to determine the synergistic effect. RESULTS: The results demonstrated a high efficacy of the two plants studied on both nematode eggs and L3 larvae as well as of their combinations. The highest egg hatching inhibition was obtained with a 50/50 combination, and the best larvae mortality was obtained with 25% A. cina and 75% T. lucida at 10 mg/mL. Additionally, this combination showed a synergistic effect. CONCLUSION: The two plant species studied here can be applied as natural anthelmintic alternatives due to their high bioactive effect and synergistic response.

Synergistic effects of using sodium hypochlorite (bleach) and desiccation in surface inactivation for Toxocara spp.

Toxocara cati and T. canis are parasitic nematodes found in the intestines of cats and dogs respectively, with a cosmopolitan distribution, and the potential for anthropozoonotic transmission, resulting in human toxocariasis. Spread of Toxocara spp. is primarily through the ingestion of embryonated eggs contaminating surfaces or uncooked food, or through the ingestion of a paratenic host containing a third-stage larva. The Toxocara spp. eggshell is composed of a lipid layer providing a permeability barrier, a chitinous layer providing structural strength, and thin vitelline and uterine layers, which combined create a biologically resistant structure, making the Toxocara spp. egg very hardy, and capable of surviving for years in the natural environment. The use of sodium hypochlorite, household bleach, as a disinfectant for Toxocara spp. eggs has been reported, with results varying from ineffective to limited effectiveness depending on parameters including contact time, concentration, and temperature. Desiccation or humidity levels have also been reported to have an impact on larval development and/or survival of Toxocara spp. eggs. However, to date, after a thorough search of the literature, no relevant publications have been found that evaluated the use of sodium hypochlorite and desiccation in combination. These experiments aim to assess the effects of using a combination of desiccation and 10% bleach solution (0.6% sodium hypochlorite) on fertilized or embryonated eggs of T. cati, T. canis, and T. vitulorum. Results of these experiments highlight the synergistic effects of desiccation and bleach, and demonstrate a relatively simple method for surface inactivation, resulting in a decrease in viability or destruction of T. cati, T. canis and T. vitulorum eggs. Implications for these findings may apply to larger scale elimination of ascarid eggs from both research, veterinary, and farming facilities to mitigate transmission.