Ultrastructural and molecular characterization of Glugea sp. (microsporidia), a parasite of the Red Sea fish Carangoides bajad (Carangidae).

Glugea sp. found infecting the liver of the teleost fish Carangoides bajad from the Red Sea, Egypt, is described based on light microscopy and ultrastructural characteristics combined with phylogenetic analyses. This microsporidium forms whitish xenomas up to ~4 mm in size. Xenomas display numerous parasitophorous vacuoles totally filled by mature spores, no other life cycle stages were observed. Mature spores ellipsoidal and measuring 6.3 × 4.0 µm in size. The polaroplast appears composed of two distinct regions: an electron-dense vesicular region and a densely packed lamellar region. The polar tubule forms approximately 24-27 coils arranged in three layers encircling the posterior vacuole. The small subunit (SSU) rRNA gene and its ITS region were sequenced and showed the highest similarity of 99.4% to other Glugea spp. Bayesian inference and maximum likelihood analyses place the novel isolate within the Glugea clade, more specifically within a subclade that predominantly grouped species described from fish inhabiting the Arabian Gulf or Red Sea. The results validate the parasite's classification in the Glugea genus. Nevertheless, until more detailed ultrastructural and molecular data are obtained, the identification of the current Glugea species is hampered by the absence of some developmental stages and the high degree of genetic similarity.

An investigation of the acaricidal activity of benzyl alcohol on Rhipicephalus annulatus and Rhipicephalus sanguineus and its synergistic or antagonistic interaction with commonly used acaricides.

The most economically significant ectoparasites in the tropics and subtropics are ixodid ticks, especially Rhipicephalus annulatus and Rhipicephalus sanguineus. Years of extensive use of the readily available acaricides have resulted in widespread resistance development in these ticks, as well as negative environmental consequences. Benzyl alcohol (BA) has been frequently used to treat pediculosis and scabies, and it may be an effective alternative to commonly used acaricides. The main aim of the present study was to evaluate the acaricide activity of BA and its combination with the regularly used chemical acaricides against R. annulatus and R. sanguineus. Different concentrations of BA alone and in combination with deltamethrin, cypermethrin and chlorpyrifos were tested in vitro against adult and larvae of both tick species. The results showed that BA is toxic to R. annulatus and R. sanguineus larvae, with 100% larval mortality at concentrations of ≥50 mL/L, and LC50 and LC90 attained the concentrations of 19.8 and 33.8 mL/L for R. annulatus and 18.8 and 31.8 mL/L for R. sanguineus, respectively. Furthermore, BA in combination with deltamethrin, cypermethrin and chlorpyrifos exhibited synergistic factors of 2.48, 1.26 and 1.68 against R. annulatus larvae and 1.64, 11.1 and 1.14 against R. sanguineus larvae for deltamethrin + BA, cypermethrin + BA and chlorpyrifos + BA, respectively. BA induced 100% mortality in adult R. annulatus at concentrations of ≥250 mL/L with LC50 and LC90 reached the concentrations of 111 and 154 mL/L, respectively. Additionally, BA had ovicidal activity causing complete inhibition of larval hatching at 100 mL/L. The combination of BA with deltamethrin and cypermethrin increased acetylcholinesterase inhibition, whereas the combination of BA with chlorpyrifos decreased glutathione (GSH) activity and malondialdehyde levels. In the field application, the combination of BA 50 mL/L and deltamethrin (DBA) resulted in a significant reduction in the percentage of ticks by 30.9% 28 days post-treatment when compared with groups treated with deltamethrin alone. In conclusion, BA causes mortality in laboratory and field studies alone and in combination with cypermethrin or deltamethrin. BA can be used for control of ticks of different life stages, that is, eggs and larvae, through application to the ground.

Repellency, Fumigant Toxicity, Antifeedent and Residual Activities of Coridothymus capitatus and Its Main Component Carvacrol against Red Flour Beetle.

Tribolium castaneum is a challenging pest of stored products, causing significant economic losses. The present study explored the efficacy of Coridothymus capitatus essential oil and its primary constituent, carvacrol, as eco-friendly alternatives for managing this pest. To evaluate their insecticidal potential, repellency, fumigant toxicity, and antifeedant properties, progeny inhibition assays were performed. Carvacrol exhibited superior repellency compared to the essential oil, achieving a 92% repellency rate at 2 mg/cm2. Both compounds demonstrated significant fumigant toxicity against T. castaneum, with LC50 values of 168.47 and 106.5 µL/L for the essential oil and carvacrol, respectively, after 24 h. Carvacrol also outperformed the essential oil in antifeedant activity, inducing an 80.7% feeding deterrence at 1.17 mg/g. Moreover, both treatments effectively suppressed the development of the pest's progeny. These results collectively underscore the potent insecticidal properties of C. capitatus essential oil and carvacrol, particularly carvacrol, as promising candidates for the sustainable management of T. castaneum in stored product protection.

D-limonene nanoemulsion: lousicidal activity, stability, and effect on the cuticle of Columbicola columbae.

The current study was conducted to investigate the efficacy and stability of D-limonene (DL) and its nanoemulsion (DLN) against pigeon feather lice (Columbicola columbae) and their mode of action. DL pure form and DLN were prepared and characterized freshly and after storage for 50 days. In vitro bioassay on live lice was conducted with different concentrations of DL, DLN, and deltamethrin (DM). The results revealed significant mortality rates in the DL-, DLN-, DM-treated groups when compared with the control (p < 0.05). The scanning electron micrographs of lice treated with DL and DLN revealed collapsed bodies with destruction in the cuticle of the mouthparts and damaged antennae. The 50 days stored DLN showed stability in their effectiveness when compared with the freshly prepared formulation. DL and DLN caused significant inhibition (p ≤ 0.05) in acetylcholinesterase activity (AchE). Malondialdehyde level (MDA) was significantly increased while glutathione was significantly decreased in DL- and DLN-treated lice. In conclusion, DL and DLN have significant lousicidal activities. DLN showed better stability than DL after storage for 50 days. In addition, the mode of action of DL may associate with its effect on the cuticle of the lice body, inhibition of AchE, and increasing oxidative stress in the treated lice.

Administration of Ethanolic Extract of Spinacia oleracea Rich in Omega-3 Improves Oxidative Stress and Goblet Cells in Broiler Chickens Infected with Eimeria tenella.

This study investigated the anticoccidial activity of spinach (Spinacia oleracea) whole-plant extract against Eimeria tenella, both in vitro and in vivo. For this purpose, one hundred 8-day-old broiler chicks of both sexes were divided into four groups (n = 25 in each group). Chicks in the first group served as the negative control (non-treated-non-infected). Chicks in the second group were challenged at 18 days old with 5 × 104E. tenella sporulated oocysts. The third group was challenged with 5 × 104 sporulated E. tenella oocysts at 18 days old after receiving spinach extract at a dose of 50 mg/kg at 8 days old. The fourth group received 0.2 mg/kg diclazuril (Coxiril® 0.2%) in their diet two days before being orally infected with 5 × 104 sporulated E. tenella oocysts and this continued till day 10 post-infection (PI). The growth performance, clinical symptoms, oocyst shedding, histological findings, and biochemical parameters were used to evaluate the efficacy on day 8 PI when the infection was at its peak. A gas chromatography examination revealed that omega-3 fatty acids were the main constituents of the spinach extract, followed by oleic acid, palmitic acid, and phytol, with amounts of 23.37%, 17.53%, 11.26%, and 7.97%, respectively. The in vitro investigation revealed that the spinach extract at concentrations of 10% and 5% inhibited the oocyst sporulation by 52.1% and 45.1%, respectively. The 5% concentration was selected for the in vivo trial based on the results of the in vitro study. The infected-untreated group showed high levels of OPG; lower body weight; a greater number of parasite stages; few goblet cells; decreased SOD, CAT, and GPX levels; and increased MDA and NO levels. The spinach-treated group, on the other hand, showed a significant decrease in oocyst output per gram of feces (OPG), increased body weight, decreased parasitic stages, and a nearly normal number of goblet cells. Additionally, it reduced malondialdehyde (MDA) and nitric oxide (NO), while increasing superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX). In conclusion, spinach produced significant antioxidant effects, increased body weight, reduced the number of oocysts and parasite stages in the caecum, and restored the number of goblet cells relative to those of an uninfected control. Furthermore, spinach extract inhibits the sporulation percentage of E. tenella oocysts. The ethanolic extract of S. oleracea (whole plant) contained high concentrations of fatty acids, palmitic acid, Phytol, betulin, and ursolic aldehyde, all of which are known to regulate the antioxidant pathway and modulate inflammatory processes and may be the main reason for its anticoccidial activity.

Toxicity and Repellency Efficacy of Benzyl Alcohol and Benzyl Benzoate as Eco-Friendly Choices to Control the Red Flour Beetle Tribolium castaneum (Herbst. 1797).

Tribolium castaneum is a damaging pest of stored grains, causing significant losses and secreting lethal quinones, which render the grains unfit for human consumption. Chemical insecticides are the most commonly used approach for control; however, they create insecticide resistance and affect the health of humans, animals, and the environment. As a result, it is critical to find an environmentally friendly pest-management strategy. In this study, two naturally occurring chemicals, benzyl alcohol (BA) and benzoyl benzoate (BB), were investigated for insecticidal activity against T. castaneum using different assays (impregnated-paper, contact toxicity, fumigant, and repellency assays). The results showed that BA had a significant insecticidal effect, with the LC50 achieved at a lower concentration in the direct-contact toxicity test (1.77%) than in the impregnated-paper assay (2.63%). BB showed significant effects in the direct-contact toxicity test, with an LC50 of 3.114%, and a lower toxicity in the impregnated-paper assay, with an LC50 of 11.75%. Furthermore, BA exhibited significant fumigant toxicity against T. castaneum, with an LC50 of 6.72 µL/L, whereas BB exhibited modest fumigant toxicity, with an LC50 of 464 µL/L. Additionally, at different concentrations (0.18, 0.09, 0.045, and 0.0225 µL/cm2), BA and BB both showed a notable and potent repelling effect. BA and BB significantly inhibited acetylcholinesterase, reduced glutathione (GSH), and increased malondialdehyde (MDA) in treated T. castaneum. This is the first report of BA insecticidal activity against the red flour beetle. Also, the outcomes of various assays demonstrated that the application of BA induces a potent bio-insecticidal effect. BA may be a promising eco-friendly alternative to control T. castaneum due to its safety and authorization by the EFSA (European Food Safety Authority).

Synergistic larvicidal and repellent effects of essential oils of three Origanum species on Rhipicephalus annulatus tick.

Ticks are of great economic importance worldwide, both because they represent major obstacles to livestock productivity and because of their ability to transmit diseases to humans and animals. Although synthetic acaricides are the most common method for tick control, their overuse has led to the development of resistance as well as unacceptable residual levels in animal products and in the environment in general. There is therefore an urgent need to identify alternative treatments. Among such alternative approaches for tick control is plant essential oil (EO) therapy. In the present study, we investigated the synergistic effect of EOs of three oregano species-Origanum onites, O. majorana and O. minutiflorum-against Rhipicephalus annulatus larvae. Gas chromatography-mass spectrometry profiles of the three EOs revealed that carvacrol was their major component, with a concentration of 86.2% in O. majorana, 79.1% in O. minutiflorum and 77.4% in O. onites. The results of larvicidal assays revealed that the doses that lead to the death of 50% of the ticks (LC50) were 22.99, 25.08 and 27.06 µL/mL for O. majorana, O. minutiflorum and O. onites EOs, respectively, whereas the doses that lead to the death of 99% (LC99) were 41.26, 43.62 and 48.96 µL/mL. In addition, the LC50 and LC99 of the three oils combined was lower (viz., 4.01 and 6.97 µL/mL) than that of each oil alone. The tested EOs were also able to repel larvae of R. annulatus to varying degrees, with O. onites oil exhibiting the greatest repellent effect, as shown by the lowest RC50 dose, followed by O. minutiflorum and O. majorana. Interestingly, this means that the oil that was least effective in killing the larvae was the most effective in repelling them. The calculated synergistic factor of any combination was higher than 1 which means that combinations have a synergistic effect. In conclusion, the combination of all three oils showed higher toxic and repellent activities than either oil separately or combinations of any two oils, suggesting synergistic effects with low doses. Further studies including field trials and the establishment of the mode of action and side effects are urgently needed to expand on these findings, and other tick stages such as adults should also be tested.

Role of antioxidant activity of essential oils in their acaricidal activities against Rhipicephalus annulatus.

Essential oils of Origanum majorana and Satureja thymbra as well as carvacrol are natural products that are known to have potent antioxidant activities. The current study was designed to investigate the role of the antioxidant properties of these natural products in their acaricidal activities against Rhipicephalus annulatus larvae. The synergistic and/or antagonistic effects of the addition of vitamins E and C and hydrogen peroxide (H2O2) to these natural products were also evaluated. Larval packet tests were used to evaluate the acaricidal activities against the larvae of R. annulatus. The antioxidant effectiveness of these products was determined by a DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging assay. The addition of vitamin E at 100 mg/mL to O. majorana and S. thymbra decreased the concentrations required to achieve the death of half of the larvae (LC50) to 0.44 and 0.47%, respectively. The combination of O. majorana and S. thymbra attained the LC50 at 1.54% which was decreased to 0.69% after addition of vitamin E. Also, the addition of vitamin E to carvacrol reduced the LC50 to 0.27%. The total antioxidant activity of these natural products increased significantly in presence of vitamin E. The addition of H2O2 inhibited the acaricidal activity of all tested materials, especially at low concentrations. All treatments induced an increase in lipid peroxidation, whereas carvacrol-treated larvae revealed the lowest values for the superoxide dismutase. Glutathione peroxidase and catalase activity decreased in larvae treated with S. thymbra combined with vitamin E. In conclusion, the addition of vitamins E and C increased the acaricidal activities of the tested compounds, whereas the addition of H2O2 decreased these activities. The antioxidant activities of essential oils and their active components may play an important role in mediating their acaricidal activities.

Correction to: Oral inoculation of ultraviolet-irradiated Eimeria species oocysts protects chickens against coccidiosis.

The author name Salama Abohamra in the original published version of this article should have been Salama Abohamra Sayed Shany.

Toxicity, repellency and anti-cholinesterase activities of thymol-eucalyptus combinations against phenotypically resistant Rhipicephalus annulatus ticks.

The current study was designed to compare the acaricidal activity of pure thymol (T) crystals, eucalyptus oil (E), and their combinations (T + E) at concentrations ranging from 0.625 to 10% against phenotypically deltamethrin-resistant Rhipicephalus annulatus ticks. The adulticidal, larvicidal, ovicidal, and repellent activities of the tested compounds and their possible mechanism of action were studied. Thymol showed 93.3% adulticidal activity at 10% concentration, whereas at 2.5% it achieved 100% larvicidal activity. Eucalyptus oil had a significant effect on tick adults at concentrations > 5%, and at 10% concentration it showed 96.7% adulticidal activity and stopped the egg hatching. On deltamethrin-resistant larvae, the median lethal concentration (LD50) was 0.61, 7.5, and 0.61% for T, E, and T + E combinations, respectively, and repellent activities were 85.7, 62.5, and 100% at the 10% concentration used. Combinations of T + E did not induce additional effects as adulticidal and larvicidal in a comparison with T treatments. The tick's acetylcholinesterase significantly decreased as a result of all treatments including deltamethrin, whereas lipid peroxidation significantly increased compared with the control untreated ticks. The glutathione content significantly decreased as a result of all treatments except for treatments with 10% thymol and its combination with eucalyptus oil. Moreover, 10% eucalyptus oil significantly decreased the tick's glutathione peroxidase and catalase activities reflecting the attenuation of antioxidant defense in the ticks. In conclusion, thymol and eucalyptus oil showed excellent activities against deltamethrin-resistant R. annulatus isolates. Their action may be mediated via acetylcholinesterase inhibition. The effect of thymol-eucalyptus combinations recorded higher activities as an ovicidal and as a repellent.

Oral inoculation of ultraviolet-irradiated Eimeria species oocysts protects chickens against coccidiosis.

Prevention of coccidiosis is one of the best ways of controlling disease. Therefore, the present study was carried out to evaluate the protective effect of ultraviolet (UV)-irradiated sporulated oocysts of Eimeria species against coccidiosis in layer chickens. One hundred forty-four one-day-old layer chicks were randomly divided into 4 groups (n = 36), including non-immunized/non-challenged negative control group (NC group), non-immunized/challenged control group (NIC group), non-irradiated sporulated oocyst/challenged group (CA group), and UV-irradiated sporulated oocyst/challenged (UV group). At the age of 4 days, chickens in groups UV and CA were both orally inoculated with 1.0 × 104 UV-irradiated and non-irradiated sporulated oocysts of Eimeria species, respectively. Chickens in groups NIC and NC were served as positive and negative controls, respectively. Chickens in all groups were orally challenged with 7.5 × 104 sporulated oocysts of Eimeria species except the NC group at the age of 21 days. The results revealed that chicks receiving UV-irradiated sporulated oocysts had no signs of illness with minimal or no changes in the cecal integrity and a significantly lower oocyst shedding (OPG) than in the NIC group. Additionally, the cytokine gene expression profiles were evaluated. Expression levels of IL-2, IL-12, and IFN-γ were significantly higher in the spleen of chicks in the UV and CA groups than in the NC group post-challenge. As expected, treatment with irradiated oocysts resulted in a significant reduction in oocyst shedding and maintenance of cecal mucosal integrity. Furthermore, the body weight was higher in chickens inoculated with UV-irradiated oocysts than their non-irradiated counterparts. In conclusion, our results demonstrate that inoculation with UV-irradiated sporulated oocysts of Eimeria species can produce a substantial reduction in infection symptoms.

Gene expression, oxidative stress and apoptotic changes in rabbit ileum experimentally infected with Eimeria intestinalis.

Coccidiosis is a parasitic disease caused by protists (apicomplexans) of the genus Eimeria Schneider, 1875 and is considered to be the most important disease faced by rabbit breeders due to its high morbidity. In the present study, the antioxidant status and changes in apoptosis and in the expression of some genes were quantified in rabbits' ilea following infection with Eimeria intestinalis Cheissin, 1948. Rabbits, orally infected with 1 × 105 sporulated oocysts of E. intestinalis, started to shed oocysts in their faeces on 8 days post infection (dpi) and reached maximum excretion on 10 dpi, with approximately 5 million oocysts. This was accompanied by a significant decrease in the live body weight of infected rabbits. Also, malondialdehyde and nitric oxide were significantly increased while catalase and glutathione were significantly decreased in the ileum tissues of the infected rabbits. In addition, a significant increase was observed in the percentages of apoptotic cells in the ilea of the infected rabbits. Furthermore, interleukin-1ß and interleukin-2 mRNA levels were significantly down-regulated and mRNA levels of interleukin-6, interferon gamma and inducible nitric oxide synthase were significantly up-regulated, while those of C-reactive protein remained unchanged. We conclude that infection with E. intestinalis induces oxidative stress, a significant increase in the percentage of apoptotic cells and a diverse and robust Th1 and Th1-related cytokine response in the ileum tissues.

Molecular diagnosis of Eimeria stiedae in hepatic tissue of experimentally infected rabbits.

The early detection of Eimeria stiedae in the hepatic tissue of experimentally infected rabbits was investigated using molecular assay. Forty 6-week-old male New Zealand rabbits were divided into two groups. Group A (30 animals) was infected with 2.5 × 10(4) sporulated oocysts of E. stiedae per animal on Day 0 and Group B (10 animals) was used as the uninfected controls. Three animals from Group A and one from Group B were sacrificed at 0, 3, 6, 9, 12, 15, 18, 21, 24 and 27 days post infection (PI). Gross and microscopic post-mortem findings were recorded. Polymerase chain reaction (PCR) of the E. stiedae internal transcribed spacer 1 genomic region was conducted on blood, liver tissue, and feces from the Group A experimentally infected animals. Macroscopically, the liver showed irregular yellowish white nodules pathognomonic to E. stiedae infection beginning on Day 15 PI. Hepatomegaly and ascites were obvious from Day 21-24 PI. The presence of different E. stiedae schizonts and gametocytes in the histopathological sections of the biliary epithelium were evident on Day 15 PI. The E. stiedae PCR was first positive in liver tissues on Day 12 and in fecal samples on Day 18 PI, but the blood samples were negative. In conclusion, the PCR can be used for early diagnosis and control of E. stiedae schizonts before shedding of the oocysts in feces.

Antioxidant properties of D-limonene and its nanoemulsion form enhance its anticoccidial efficiency in experimentally infected broilers with Eimeria tenella: an in vitro and in vivo study.

The excessive use of conventional medications to treat coccidiosis has led to concerns regarding drug residues in tissues and the emergence of multidrug resistance. Essential oils with anti-inflammatory and antioxidant activities may also have anticoccidial effects. The present study investigated the efficacy of D-limonene and its nanoemulsion form against Eimeria tenella in chickens. An in vitro study was conducted to evaluate the sporulation inhibitory effects of D-limonene on Eimeria tenella oocysts. Five D-limonene concentrations (0.625, 1.25, 2.5, 5, and 10% v/v) were tested alongside positive (10% formalin) and negative (2.5% potassium dichromate) controls. Each ELISA plate well was inoculated with 1200 unsporulated oocysts and incubated at 30 °C for 24, 48, and 72 h. Subsequently, samples were microscopically examined to assess sporulation inhibition and calculate the percentage of sporulated oocysts. For the in vivo study, 125 eight-day-old broiler chicks were divided into five groups of 25 birds each. The control negative group remained uninfected and untreated. The control positive group was challenged with 5 × 104 sporulated Eimeria tenella oocysts. The diclazuril group received 0.2 mg/kg diclazuril in their diet two days prior to, and until 10 days post infection. The D-limonene (DL) and D-limonene nanoemulsion (DLN) groups were challenged with 5 × 104 sporulated E. tenella oocysts at 18 days of age and administered 150 mg/L of their respective treatments in drinking water from day eight until the end of the experiment. Results from the in vitro study demonstrated that D-limonene suppressed oocyst sporulation by 50.83% at its highest concentration of 10%. In the in vivo study, both DL and DLN treated groups exhibited a significant reduction in oocyst output per gram of feces (OPG), along with increased body weight and decreased parasite stages in the cecal tissue. Furthermore, these treatments were associated with elevated levels of antioxidant enzymes such as glutathione peroxidase (GPX), catalase (CAT), and superoxide dismutase (SOD), accompanied by a decrease in malondialdehyde (MDA) and nitric oxide (NO) levels. Particularly, DLN treatment remarkably increased the number of goblet cells. In conclusion, D-limonene and its nanoemulsion represent promising alternatives for managing coccidiosis in poultry. They not only effectively control parasites but also promote intestinal health and boost antioxidant defenses.

Efficacy and safety of geranium-oregano-thymol formulations to control of dog tick Rhipicephalus sanguineus sensu lato under laboratory and field conditions.

The present study evaluated, in laboratory and field, the efficacy and safety of formulations of Pelargonium graveolens (geranium - G), Origanum majorana (oregano - O) commercial essential oils (EO) and thymol (T) to control of Rhipicephalus sanguineus sensu lato. In the laboratory, three formulas (A: 2% tween 80%, B: powder and C: nanoemulsion) by a mixture of these components (GOT) were prepared and evaluated, and the best one was used to assess its safety and field application against R. sanguineus s. l. on naturally infested dogs. Besides the major compounds of the EO used were identified. The results of the lab study showed that formula A (2.5 g of each G + O + T + 2% tween 80 to complete 100 mL) was significantly more effective than the other two formulas tested and exhibited highly effective adulticidal, larvicidal, and ovicidal activity against R. sanguineus s.l. Significant LC50 and LC90 values of GOT were evaluated (13.4 and 21.5 mg/mL, respectively) for the adulticidal activity, (2.81 and 4.46 mg/mL, respectively) for ovicidal activity and (2.44 and 4.45 mg/mL, respectively) for larvicidal activity. The safety of formula A has been proven by the absence of its cytotoxicity on a cell line of human epidermoid carcinoma. Citronella and carvacrol were the major compounds identified in the commercial essential oils of P. graveolens and O. majorana, respectively. Formula A was used in a field control trial for almost 8 months, during the tick infestation season (April to November, 2022). Fourteen naturally infested dogs were divided into two groups, each with seven dogs. One group received formula A spraying five times during an experiment that continued for 8 months, while the other group received treatment with commercially available malathion acaricide. The animals were sprayed on five occasions throughout the experiment (April, June, July, August, and September). The results showed a substantial percentage of effectiveness after the first application of formula A with a 99.3% reduction in tick count at day 28 post-application (PA). In the case of severe infestation 60 days after the first application of formula A (more than 180 ticks per dog), the second application was done, achieving an efficacy of 54.9% at day 3 PA, so an emergency spray was done at day 5 PA to combat the rest of the tick infestation, achieving efficacy of 99% after 3 days. Consequently, a regular spray (third, fourth, and fifth application) was done every 35 days. This regular spray revealed 100% effectiveness at 14 days PA. Biochemical parameters of treated dogs were evaluated to confirm the safety of formula A. Creatinine, ALT, and albumin of the dogs treated with formula A were within the normal range of dogs, while urea and AST were higher than the normal range. In conclusion, formula A can safely treat R. sanguineus s.l. infestations in dogs with regular application every 5 weeks.

Benzoate Derivatives Toxicity to Musca domestica Results in Severe Muscle Relaxation and Body Distortion.

The house fly, Musca domestica (Linnaeus) (Diptera: Muscidae), is a significant threat to human and animal health and is also resistant to a variety of insecticides. Plant-derived benzoates are known to have insecticidal activities against various insects. In this study, the larvicidal, pupicidal, and adulticidal activities of benzoate derivatives (benzyl alcohol BA, benzyl benzoate BB, and methyl benzoate MB) were assessed and investigated for their effects on larval structure and acetylcholinesterase activity. Six concentrations (2.5 to 100 mg/mL) of benzoate derivatives were applied to larvae and pupae through the residual film method and topical application, respectively. Meanwhile, concentrations from 0.625 to 50 mg/L air were applied to adult flies through a fumigation assay. BA and MB achieved promising results against larvae with LC50 values of 10.90 and 11.53 mg/mL, respectively. Moreover, BA killed 100% of the larvae at a concentration of 25 mg/mL, and MB achieved the same effect at a concentration of 50 mg/mL. Regarding the pupicidal activity, MB showed a percentage inhibition rate (PIR) of 100% at a concentration of 100 mg/mL, while the same effect was achieved by BA at a concentration of 50 mg/mL. Meanwhile, BB did not show any effect on the larvae or pupae at any of the tested concentrations. Moreover, the scanning microscopy observations on the treated larvae by BA and MB estimated flaccid and deformity in the larva body with a shrunken cuticle. Additionally, both BA and MB suppress nerve signal transmission by inhibiting acetylcholinesterase. In conclusion, the results of this study indicate that BA and MB may be useful in control housefly populations. These substances cause severe muscular relaxation and deformities in insects.

Frequency of Besnoitia besnoiti and Neospora caninum antibodies in cattle and small ruminants from greater Cairo and Beni Suef governorates, Egypt.

Livestock industry is a pivotal sector for providing food, work places and monetary support for Egyptian people. Besnoitia besnoiti and Neospora caninum are protozoan parasites that are responsible for significant economic losses in ruminants, particularly in cattle. Besnoitia besnoiti can cause fertility problems and a general loss in productivity, while N. caninum is a major cause of abortion and neonatal abnormalities in infected animals. There is little information on the existence of these protozoa in Egypt, thus we conducted this study to reveal the current situation in cattle (n = 264), sheep (n = 151), and goats (n = 25). Serum samples were collected from governorates of Cairo, Giza, and Beni Suef, representing the most densely populated regions in Egypt. Using commercial ELISAs, an overall estimation among all tested animals (n = 440) revealed 7.7%, 13.2%, and 0.9% as seropositive rates for B. besnoiti, N. caninum, and mixed infection, respectively. Animal species (cattle vs sheep vs goat) and age of cattle (less than vs >1 year old) were analyzed as risk factors for infection. Regarding B. besnoiti, the seroprevalence was significantly higher in cattle than in sheep and goats and in adult cattle than calves. For N. caninum infection, no significant differences were recorded, although the seropositive rates were higher in cattle, and in adult cattle. This study provides the first seroprevalence data for B. besnoiti in all surveyed animals in the regions included, and in sheep and goats from Egypt, and supports the current knowledge for the occurrence of N. caninum in Egypt.

Efficacy of D-Limonene Nanoemulsion Against Rhipicephalus annulatus and Rhipicephalus sanguineus Ticks.

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.

Safety and Efficacy of Pure and a Nanosuspension of D-limonene for Controlling Pigeon Lice.

This study investigated the safety and efficacy of two forms of D-limonene (DL) against Columbicola columbae (pigeon feather lice); pure and a nanoemulsion formulation (DLN). The cell cytotoxicity of the prepared forms of DL/DLN was investigated using skin cell lines. In vitro and ex vivo bioassays were applied on lice. The ex vivo bioassay was done on cut feathers containing lice eggs. The in vivo experiment was conducted on pigeons naturally infested by lice. The infested pigeons were treated with DL, DLN, or deltamethrin (D) as a positive control. Both forms of D-limonene were found to be safe when applied to the normal human skin fibroblast cell line, but DLN was toxic to skin cell carcinoma. The in vitro and ex vivo results of both DL and DLN forms were similar. All eggs treated with DL, DLN, and D failed to hatch (100%). The in vivo results showed complete elimination of lice 24 h post-treatment (PT), and biochemical analysis showed that the treated birds retained normal kidney and liver functions. Treated groups also showed improved productivity in the 4 months PT. In conclusion, DL and DLN are safe and effective in controlling feather lice infestation in pigeons and successful treatment encourages bird productivity.