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Ivermectin (Iv) is widely used to control Rhipicephalus annulatus infestations in cattle in Egypt. However, increasing resistance to ivermectin necessitates alternative approaches. This study aimed to address this resistance by investigating the interactions between ivermectin and essential oil components (EOCs)-carvacrol, cinnamaldehyde, D-limonene, trans-anethole, and thymol-against four R. annulatus tick isolates collected from Beni-Suef, El-Minia, and El-Fayoum. The larval immersion test was conducted using various ivermectin concentrations (3, 1.5, 0.75, 0.38, 0.19, 0.09, 0.045, 0.025, and 0.0125 mg/mL) prepared in EthTX1 %. Additionally, EOCs were tested at different concentrations (20, 10, 5, and 2.5 mg/mL) prepared by dilution in 5 % ethyl alcohol and EthTX 1 %. Ivermectin and each EOC were tested individually against the isolates. Subsequently, the LC50 of ivermectin was combined with the LC50 of each EOC in a 1:1 ratio to evaluate potential synergistic effects. All EOCs demonstrated significant toxicity against the isolates. The combination of carvacrol and thymol with ivermectin increased mortality, but the combination index (CI) exceeded one for all isolates, indicating no synergism. Similarly, while D-limonene, cinnamaldehyde, and trans-anethole also resulted in high mortality rates, including up to 100 %, no synergism was observed (CI > 1). However, these three EOCs exhibited CIs close to 1, suggesting an additive effect when combined with ivermectin. In conclusion, combining ivermectin with certain EOCs, particularly D-limonene, cinnamaldehyde, and trans-anethole, may offer an effective tool for controlling cattle ticks. Developing topical or pour-on formulations of ivermectin-EOC combinations could be an integral part of integrated management strategies to combat resistance to macrocyclic lactones like ivermectin. Further studies, including tests on adult ticks and field trials, are needed to evaluate the efficacy of these combinations., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships., (Copyright © 2024 Elsevier B.V. All rights reserved.)

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 LC 50 and LC 90 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., Competing Interests: Declaration of Competing Interest The authors declare that the research was conducted in the absence of any commercial or financial relationships., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Haemonchosis is a worldwide helminthic disease affecting ruminants. The anthelminthic resistance has become raised. Medicinal plants are safely used as synthetic anthelmintics. Currently, the efficacy of black pepper essential oil (BPO) and tea tree oil (TTO) were in vitro evaluated against Haemonchus contortus adults, eggs and larvae at concentrations of 1.25, 2.5 and 5 mg/ml in addition to the commercially used albendazole at a concentration of 10 μg/ml. Oils were used in both normal and nanoparticles-loaded forms. Oxidative stress enzymes of worms were estimated. Scanning electron microscopy (SEM) for treated worms was done. Both normal and nanoemulsion forms of both BPO and TTO stopped the adult motility [BPO 2.5 h (hrs), NBPO 1.5 h, TTO 3 h, NTTO 1.5 h] and induced a marked decrease in the oviposition. Post treatment, the egg development and hatching were significantly ( P  ≤ 0.05) reduced. The damage of the egg shell, embryonal cessation and destruction of larvae occurred. Noticeable elevated antioxidant enzymes (catalase CAT, glutathione transferase GST and glutathione GSH) were found, while oxidative enzymes (lipid peroxidation LPO and nitric oxide synthase NOS) decreased. Scanning electron microscopy (SEM) for both oil-treated worms revealed anterior ends damage and several cuts associated with cuticular pores. The use of albendazole induced more or less anthelmintic and enzymatic activities with less morphological alterations of adults revealed by SEM. This study proved the marked anthelmintic potency of the BPO and TTO and their nanoemulsion forms against H. contortus rather than the widely used anthelmintic drugs., Competing Interests: Conflict of interestNo conflict of interest in the submission of this manuscript, and manuscript is approved by all authors for publication., (© Indian Society for Parasitology 2024. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

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 LC 50 and LC 90 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 LC 50 and LC 90 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., (© 2023 Royal Entomological Society.)

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.)

Cysticercus bovis is the metacestode of the commonly defined foodborne cestode, Taenia saginata . It infects heart, masseter muscles and other muscular sites of cattle causing bovine cysticercosis. So, a retrospective study using archival documents from authorized agencies is done during the period of 2018-2020 with estimating economic losses. Cysts were collected from municipal Beni-Suef and Basatin abattoirs. Molecular approach including cPCR and sequencing analysis is used to prove the species. The prevalence was 0.34% in Beni-Suef abattoir and 5.70% in Basatin abattoir including heads, hearts or all carcasses condemned. Among 27 provinces surveyed, the highest prevalence was recorded in Aswan followed by Cairo, Suez and Red Sea. Moreover, 19 provinces (Cairo, Alexandria, Suez, Dakahlia, Sharkia, Qalyobia, Kafrelsheikh, Gharbia, Menofiya, Ismailia, Giza, Beni-Suef, Assiut, Sohag, Qena, Aswan, Red Sea, South Sinai and El-Wadi El-Gadid) recorded bovine cysticercosis in 2018, 2019 and 2020. On the other hand, Luxor and North Sinai had no infections. The percentages of condemnation among slaughtered cattle were 1.38, 1.49 and 0.87% in 2018, 2019 and 2020, respectively. Condemnations significantly varied among north, middle and south districts of Egypt, and also varied annually. Molecularly, a diagnostic band at an amplicon size 253 bp targeting the COI gene specific for Cycticercus bovis was revealed. The obtained sequences showed 100% identity with the different Taenia saginata COI GB sequence isolates in many countries worldwide. The phylogenetic analysis method showed that the obtained sequences originating from the same clade of Taenia saginata GB isolates globally. Careful meat inspection as well as strict hygienic measures is recommended for both veterinarians and public., Competing Interests: Conflict of interestAuthors declare that there is no conflict of interest., (© Indian Society for Parasitology 2023. Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

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., (© 2022 Royal Entomological Society.)

While the importance of cryptosporidiosis in immunocompromised persons is well known, the prevalence of Cryptosporidium spp. in cancer patients is not clear. The current study was designed to assess the occurrence and genetic characteristics of Cryptosporidium spp. in patients with gastrointestinal (GI) cancer in Egypt. Stool samples were collected from 100 patients with GI malignancies and 20 healthy individuals without any GI manifestations (control group). They were screened by microscopy and the immunochromatographic RIDA®QUICK Cryptosporidium kit. Subtyping of Cryptosporidium spp. was conducted by sequence analysis of the glycoprotein 60 (gp60) locus. Sociodemographic, environmental data and information on GI symptoms, cancer types, and clinical treatment were obtained via a questionnaire. By microscopy and RIDA®QUICK, only 7% (7/100) of GI cancer patients were positive for Cryptosporidium, compared with 40% (40/100) by gp60 nPCR. No positives were obtained from the control group. Male sex (P = 0.02) and younger age (P = 0.004) were major Cryptosporidium risk factors for infection. The occurrence of Cryptosporidium was also significantly more frequent (P = 0.003) in watery stool samples. Sequence analysis of the gp60 amplicons (~ 400 bp) identified a novel C. parvum subtype with nine TCA repeats and eleven ACATCA repeats. A formal subtype designation could not be made due to the short sequence length. More studies should be conducted to verify the common occurrence of this unusual C. parvum subtype and establish its genetic identity., (© 2022. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

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., (© The Author(s) 2022. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Background: Toxoplasma gondii and Neospora caninum are major protozoan parasites of worldwide distribution and significance in veterinary medicine and, for T. gondii , in public health. Cats and dogs, as final hosts for T. gondii and N. caninum, respectively, have a key function in environmental contamination with oocysts and, thus, in parasite transmission. Very little is known about the prevalence of T. gondii infections in dogs and cats in Egypt, and even less about the prevalence of N. caninum in the same hosts., Methods: In the current study, 223 serum samples of both dogs (n = 172) and cats (n = 51) were investigated for specific antibodies to T. gondii and N. caninum using commercially available ELISAs. A risk factor analysis was conducted to identify factors associated with seropositivity., Results & Discussion: Exposure to T. gondii was reported in 23.3% of the dogs and in 9.8% of the cats, respectively. In addition, N. caninum -specific antibodies were recorded in 5.8% of dogs and in 3.4% of cats. A mixed infection was found in two dogs (1.2%) and in one cat (2%). Antibodies to T. gondii in dogs were significantly more frequent in dogs aged 3 years or more and in male German Shepherds. As this breed is often used as watchdogs and was the most sampled breed in Alexandria governorate, the purpose "watchdog" (compared to "stray" or "companion"), the male sex, and the governorate "Alexandria" also had a significantly higher seroprevalence for T. gondii . No factors associated with antibodies to N. caninum could be identified in dogs, and no significant factors were determined in cats for either T. gondii or N. caninum infection. Our study substantially adds to the knowledge of T. gondii infection in dogs and cats and presents data on N. caninum infection in cats for the first and in dogs in Egypt for the second time.

A retrospective study to estimate economic losses caused by livers condemnation, due to fascioliasis, of slaughtered cattle and buffaloes in Egypt during the period of 2016-2020, was done. Moreover, a morpho-molecular identification of collected liver flukes from slaughtered animals in municipal abattoirs was conducted. Livers of naturally infected carcasses were obtained from slaughtered animals in Beni-Suef, Cairo and Tanta provinces, Egypt during 2019-2020 for phenotypic characterization of recovered Fasciola species and molecular identification of collected worms using PCR targeting the ITS-1 region. Findings of the retrospective study revealed that percentages of livers condemnation of cattle and buffaloes ranged from 0.79 to 0.66% during the period from 2016 to 2020. The highest percentages were detected in the south Egypt (2.5-6.0%) with the highest economic losses (261850-616300 USD annually). Morphometrically, collected flukes categorized into Fasciola hepatica and Fasciola gigantica . No intermediate forms ( Fasciola sp.-like) were detected. Sequencing analysis of ITS-1 PCR products showed that only Fasciola hepatica (26/34) and F. gigantica (8/34) isolates were found, with no intermediate forms, Fasciola sp.-like, could be identified. Currently, Fasciola hepatica was 100% identical with the Egyptian species (LC076196 and JF294998), French species (JF294999), and Iranian species (MF969009 and MK377150). Moreover, the obtained F. gigantica species showed 100% identity with Egyptian ones (LC076125, LC076108 and KX198619), Iranian (KF982047 and MF372919), and other GenBank specimens from Vietnam, Cameroon and India. In conclusion, South Egypt showed the highest economic losses due to fascioliasis, especially Aswan province. Fasciola hepatica was more common than F. gigantica , while the hybrid form was not detected., Competing Interests: Conflict of interestAuthors declare that there is no conflict of interest., (© Indian Society for Parasitology 2022, Springer Nature or its licensor holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.)

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 (H 2 O 2 ) 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 (LC 50 ) to 0.44 and 0.47%, respectively. The combination of O. majorana and S. thymbra attained the LC 50 at 1.54% which was decreased to 0.69% after addition of vitamin E. Also, the addition of vitamin E to carvacrol reduced the LC 50 to 0.27%. The total antioxidant activity of these natural products increased significantly in presence of vitamin E. The addition of H 2 O 2 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 H 2 O 2 decreased these activities. The antioxidant activities of essential oils and their active components may play an important role in mediating their acaricidal activities., (© 2022. The Author(s), under exclusive licence to Springer Nature Switzerland AG.)

Hymenolepiasis represents a parasitic infection of common prevalence in pediatrics with intimidating impacts, particularly amongst immunocompromised patients. The present work aimed to snowball the curative outcomes of the current mainstay of hymenolepiasis chemotherapy, praziquantel (PRZ), through assembly of polymeric mixed micelles (PMMs). Such innovative nano-cargo could consolidate PRZ hydrosolubility, extend its circulation time and eventually upraise its bioavailability, thus accomplishing a nanoparadigm for hymenolepiasis tackling at lower dose levels. For consummating this goal, PRZ-PMMs were tailored via thin-film hydration technique integrating a binary system of Lutrol F127 and Gelucire 44/14. Box-Behnken design was planned for optimizing the nanoformulation variables employing Design-Expert ® software. Also, in Hymenolepis nana -infected rats, the pharmacodynamics of the optimal micellar formulation versus the analogous crude PRZ suspension were scrutinized on the 1st and 3rd days after administration of a single oral dose (12.5 or 25 mg/kg). Moreover, in vitro ovicidal activity of the monitored formulations was estimated utilizing Fuchsin vital stain. Furthermore, the in vivo pharmacokinetics were assessed in rats. The optimum PRZ-PMMs disclosed conciliation between thermodynamic and kinetic stability, high entrapment efficiency (86.29%), spherical nanosized morphology (15.18 nm), and controlled-release characteristics over 24 h (78.22%). 1 H NMR studies verified PRZ assimilation within the micellar core. Additionally, the in vivo results highlighted a significant boosted efficacy of PRZ-PMMs manifested by fecal eggs output and worm burden reduction, which was clearly evident at the lesser PRZ dose, besides a reversed effect for the intestinal histological disruptions. At 50 µg/mL, PRZ-PMMs increased the percent of non-viable eggs to 100% versus 47% for crude PRZ, whilst shell destruction and loss of embryo were only clear with the applied nano-cargo. Moreover, superior bioavailability by 3.43-fold with elongated residence time was measured for PRZ-PMMs compared to PRZ suspension. Practically, our results unravel the potential of PRZ-PMMs as an oral promising tolerable lower dose nanoplatform for more competent PRZ mass chemotherapy.

The effect of thymol and ivermectin on the development and embryonation of Toxocara vitulorum ( T. vitulorum ) eggs, as well as their migration in albino rats was investigated both in vitro and in vivo. A total of forty male albino rats were divided into four groups for an in vivo experiment. The first group was uninfected; the second group was infected but left untreated; the third group was infected and received thymol at a dose of 40 mg/kg; and the fourth group was infected and received ivermectin (0.2 mg/kg). In vitro, thymol inhibited the development of Toxocara larvae within the eggs. However, ivermectin, produced inconsistent results. The in vivo results indicated that the recovery rates of Toxocara larvae from the liver and lungs on day 7 post-infection were significantly lower in the thymol or ivermectin-treated groups than in the infected untreated control. Albumin levels were significantly increased in the thymol-treated group as compared to the positive control and ivermectin groups. Nitric oxide, IL-4, and IFN- levels in the serum of the thymol or ivermectin-treated groups were significantly lower than that of the positive control group. Histopathological examination demonstrated that thymol and ivermectin were effective in reducing larval load, reducing the number and size of granulomas in the absence of larvae, and improving tissue architecture. The current study concluded that thymol possessed anti- Toxocara activity in a rat model. Additionally, thymol possessed ovicidal properties and may be used as a disinfectant., Competing Interests: Conflict of interestThe authors have declared that no competing interests exist., (© Indian Society for Parasitology 2022.)

Globally, the economic losses due to hard ticks infestation and the control of the associated diseases have been calculated at USD $13.9-18.7 billion per year. The economic impact is related to its direct damage to the skins, blood loss, anemia, severe immunological reactions and indirect losses that related to the effects of hemoparasites, cost of treatment for clinical cases and expenses incurred in the control of ticks. The current study evaluated the acaricidal activities of fennel Foeniculum vulgare essential oil and its main components; trans-anethole and fenchone; against R. annulatus. GC-MS analysis revealed that this oil contained 16 components representing 99.9% of the total identified compounds with E-anethole being the predominant component(64.29%), followed by fenchone (9.94%). The fennel oil and trans-anethole showed significant acaricidal activities. The LC50 of the fennel oil was attained at concentrations of 12.96% for adult ticks and 1.75% for tick larvae meanwhile the LC50 of trans-anethole was reached at concentrations of 2.36% for adult tick and 0.56% for tick larvae. On the contrary, fenchone showed no any significant adulticidal activities and its LC50 attained at a concentration of 9.11% for tick larvae. Regarding repellence activities, trans-anethole achieved 100% repellency at the concentration of 10% while fennel showed 86% repellency at the same concentration. Fenchone showed no repellency effect. Treatment of larvae with fennel, trans-anethole, and fenchone LC50 concentrations significantly inhibited the acetylcholinesterase activity. Meanwhile, glutathione s-transferase activity was significantly decreased in fennel treated larvae but no significant effect was found in the larvae of trans-anethole and fenchone groups. These results indicate that the acaricide effect of fennel oil may attributed to its high content of trans-anethole. This was supported by potent adulticidal, larvicidal, and repellency effects of trans-anethole against Rhipeciphalus annulatus tick and therefore it could be included in the list of acaricide of plant origin., Competing Interests: The authors have declared that no competing interests exist.

The purpose of this study was to determine the course of infection in pigeons with Eimeria labbeana . Thirty-five squabs (4-6 weeks old) were brought from the local poultry market and examined for coccidial infection for 7 days to ensure they were coccidia free. A negative control group of five squabs was used, and thirty squabs were infected orally with 2.5 × 10 4 sporulated E. labbeana oocysts. From day 1-8 post-infection (PI), three squabs were scarified daily to track the endogenous stages in the intestinal tissue. Furthermore, six squabs were preserved to track the patent period and calculate daily oocyst shedding. The parasite stages were differentiated using paraffin-embedded intestinal tissues that were sectioned and stained. On day 5 PI, the infected squabs had greenish watery diarrhea, weakness, rough feathers, and decreased food intake. The pre-patent and patent durations were six and fourteen days PI, respectively. The shedding of oocysts began on day 6 PI and peaked on day 8 PI. In the duodenum and jejunum of the small intestine, histopathological investigation indicated the presence of three schizont stages, macro- and micro-gametes, and oocysts. To the best of our knowledge, this is the first study in Egypt to explore the course of E. labbeana infection in domestic pigeons., Competing Interests: Conflict of interestThe authors declare that there is no conflict of interests regarding the publication of this article., (© Indian Society for Parasitology 2021.)

Purpose: Susceptibility and resistance of Haemonchus contortus to anthelmintic drugs, including ivermectin, levamisole, and albendazole in naturally infected sheep were investigated., Methods: Three sets of assays were conducted to detect drug efficacy. Firstly, in vivo estimation of drug resistance to H. contortus was explored in 80 sheep naturally infected with H. contortus. Sheep were divided into four equal groups (20 sheep for each group): the first group was treated with albendazole (5.00 mg/kg BW), the second with levamisole (7.50 mg/kg BW), the third with ivermectin (0.20 mg/kg BW), and the fourth group served as the untreated control. Fecal egg reduction test (FERT) was done at days 7 and 14 after treatment. Secondly, for in vitro egg hatching assay (EHA), H. contortus eggs from naturally infected sheep were collected and treated with 0.0002, 0.002, 0.02, 0.2, and 2.0 µg/mL albendazole. Thirdly, molecular detection of the albendazole resistance gene in adult male H. contortus worms and larvae from infected sheep was carried out using allele-specific PCR., Results: The FECRT results showed that the drug efficacy was 86.84% for albendazole and 100% for both levamisole and ivermectin. The result of EHA showed that eggs did not hatch at 2.0 µg/mL albendazole concentration. Molecular findings showed two forms, including H. contortus homozygous susceptible (SS) and heterozygous (RS) of "β-tubulin" gene at 200 sites, which were recorded in both single male worms and larvae., Conclusion: H. contortus, which was susceptible to levamisole and ivermectin, had developed resistance to albendazole.

The current study investigated the synergistic effect of combinations containing deltamethrin (D), Eucalyptus essential oil (E), and the thyme essential oil component thymol (T), against a field population of Rhipicephalus annulatus in Egypt that was characterized to be resistant to D. Solutions of T, E, or TE at concentrations of 1.25-5% were combined with 5% deltamethrin at different dilutions (0.25-2 mL/L). Results of the adult immersion test used to estimate the in vitro acaricidal activity of these combinations at 5% yielded LC 50 values for D, E-D, T-D, and TE-D of 3.87 mL/L, 3.89 mL/L, 0.14 mL/L, and 0.05 mL/L, respectively. Biochemical analyses using whole-body homogenate of ticks from the in vitro tests revealed that the lowest acetylcholinesterase and glutathione peroxidase activity, and the maximum lipid peroxidation were recorded in ticks treated with 5% TE-D. Glutathione content significantly decreased (p ≤ 0.05) in all treated ticks. Three groups, each containing five cross breed cattle naturally infested with R. annulatus from the same area where resistance to D was detected, were sprayed twice at two-week intervals using 1 mL/L of 5% solutions of D, T-D, or TE-D. Overall efficacy of the D, T-D, and TE-D sprays by day 30 post-treatment was 21.6, 88.3, and 95 %, respectively. Ticks collected from infested cattle three days after treatment with the D spray deposited egg masses that were able to hatch, deposited small masses of eggs unable to hatch when exposed to the T-D spray, and laid few eggs that didn't hatch when sprayed with the TE-D combination. Values for liver and kidney function parameters were comparable in cattle before and after treatment with the combination sprays tested. The TE-D spray overcame the insensitivity to D of this R. annulatus population in Egypt, which also highlighted the significant synergistic effect of thymol on the acaricidal activity of deltamethrin observed in vitro. Acaricidal activity of the TE-D combination apparently has deleterious effects on multiple tick systems involving inhibition of acetylcholinesterase, increased lipid peroxidation, and oxidative stress. These findings document that combinations of natural and synthetic products can be part of integrated management solutions to the problem with widespread resistance to pyrethroids like deltamethrin in populations of cattle ticks, including R. annulatus, around the world., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Under the poor hygienic conditions, tick-borne pathogens cause severe economic losses to the cattle industry., Purpose: The current study investigated the presence of Theileria annulata, Babesia bigemina, and Anaplasma marginale, the most relevant tick-borne pathogens in cattle, in 3 provinces of Egypt utilizing species-specific PCR assays., Methods: PCR was conducted, on bovine blood specimens, using primers targeting the T. annulata merozoite-piroplasm surface antigen (Tams1, 768 bp), A. marginale major surface protein-1b gene (msp1b, 265 bp), and B. bigemina small subunit ribosomal RNA gene (SSrRNA, 543 bp)., Results: PCR findings revealed overall prevalences of T. annulata, B. bigemina, and A. marginale as 22.0% (33/150), 19.33% (29/150), and 10.6% (16/150), respectively. The co-infection with two or three pathogens was detected in 20.0% (30/150) of examined specimens. Sequence analyses indicated that T. annulata and A. marginale varied from those of corresponding GenBank sequences revealing percent identities ranging from 90.68 to 97.75% and from 94.98 to 98.63%, respectively. On the other hand, the obtained B. bigemina sequences showed a high similarity with those previously reported in GenBank with a percent identity ranging from 98.85 to 100%., Conclusion: T. annulata was the most prevalent tick-borne pathogen in examined bovine specimens. The genetic diversity of markers used for identification of T. annulata and A. marginale should be highly considered.

Rhipicephalus annulatus field populations collected from small cattle farms in Beni-Suef province in Egypt were evaluated for deltamethrin resistance by toxicological in vitro bioassays (adult immersion test-AIT and larval packet test-LPT). Moreover, a quantitative PCR high resolution melting (PCR-HRM) technique was used to detect nucleotide substitutions in the voltage-gated sodium channel (Na-channel) gene. By the in vitro bioassays, the examined ticks were phenotypically categorized as deltamethrin susceptible (populations El-Wasta - A, and El-Hakamna - C) or resistant (populations El-Wasta - B, El-Hakamna - D, EL-Halabia - E, and Kom-abokhalad - F). Based on LPT findings, the phenotypic resistant populations were found to have a resistance ratio between 6.5 - 10.8. The PCR-HRM genotyping of the ticks showed variable melting curves among the populations in domain II of the Na-channel gene. Analysis of the curves showed the presence of wild type, mutant homozygous, and mutant heterozygous tick individuals. By sequencing the PCR amplified fragments, the C190A mutation was the only detected nucleotide polymorphism of domain II among the phenotypically resistant populations, which was present in 39.5 % (34/86) of the ticks tested. On the other hand, the phenotypically susceptible populations A and C did not show C190A mutant homozygous (RR) individuals. Meanwhile, in domain III all of the examined populations revealed melting curves like the wild type. Furthermore, the sequence analysis of these populations confirmed the absence of SNPs in domain III. The C190A single point mutation was detected for the first time in domain II of the Na-channel gene of deltamethrin-resistant R. annulatus in Egypt using PCR-HRM. Screening for efficacy of chemical compounds used by farmers to control ticks on cattle should be considered as part of animal health programs to manage the emerging resistance to acaricides in R. annulatus populations., Competing Interests: Declaration of Competing Interest The authors declare that they have no conflict of interest., (Copyright © 2020 Elsevier GmbH. All rights reserved.)

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 (LD 50 ) 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.

Investigation of thymol efficacy to control pigeon coccidiosis was performed using in-vitro and in-vivo studies. The in-vitro experiment was conducted by treatment of unsporulated oocysts of Eimeria species of pigeon by five concentrations (0.625-10%) from either thymol, eucalyptus essential oil or amprolium anticoccidial drug and incubation for 72 h. The in-vitro study revealed that thymol concentrations ≥1.25 % caused significant deformity on sporulated and unsporulated oocysts compared to the other two products. Eucalyptus oil was active at both 5 and 10 % concentrations on unsporulated oocysts but showed non-significant changes on sporulated ones at all tested concentration. Meanwhile, in-vivo testing of thymol was conducted using 45 squabs which were equally divided into three groups; untreated uninfected (UU) negative control, untreated infected (UI) positive control and thymol treated (TT). TT group received 40 mg/kg BWt thymol in feed for 15 days. At day five post thymol supplementation, the UI and TT groups were orally infected by 25 × 10 3 sporulated oocysts of pigeon Eimeria labbeana. The in-vivo study showed that thymol minimized the adverse effect of Eimeria infection in pigeon as observed by less severity of clinical signs, low oocysts count and improvement of body weight when compared with untreated infected birds. In addition, the biochemical parameters including liver and kidney functions tests proved thymol safety in pigeon. Moreover, thymol showed excellent antioxidant activity that was estimated by significantly lower value of malondialdehyde in TT than UI groups. The histopathological findings of TT group showed intact intestinal villi with mild sloughed epithelium, degenerated coccidian developmental stages and massive infiltrations of mononuclear cells in lamina propria. In conclusion, thymol can be safely used to control pigeon coccidiosis as a natural effective compound., Competing Interests: Declaration of Competing Interest The authors declare that there is no conflict of interests regarding the publication of this article., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Cystic echinococcosis is a potential zoonotic helminthic disease affect a broad spectrum of mammals including humans worldwide. The economic impact of the disease interestingly requires updated gathering information on the prevalence in slaughtered animals. Accordingly, in the current study, 573 camels, 4300 sheep, and 1235 pigs were surveyed in four Egyptian municipal abattoirs. Among those, 62 (10.82%) camels, 33 (0.77%) sheep and 3 (0.24%) pigs had cystic echinococcosis in lungs, livers and spleen. The diversity of cysts revealed that positive-cystic echinococcosis animals from all species were detected in El-Basatin abattoir. In El-Monieb abattoir, sheep only were infected. In El-Waraa and Beni-Suef abattoirs, cysts were seen in camels only. Infected animals included both sexes. In camels, lungs were the most affected organs, while in sheep, livers were the most abundant. In pigs, hydatid cysts were detected only in lungs. Moreover, camel cysts were mostly large-sized (diameter > 10 cm) with smaller cysts are also recorded. In sheep, small-, moderately- and large-sized cysts were equally present. Cysts in pigs were large-sized. Camels showed a higher percentage of fertile cysts (46.77%; 29/62) followed by sheep (21.21%; 7/33). Calcified/degenerated cysts were less in camels (38.71%; 24/62) than in sheep (75.76%). Non-viable cysts were the less frequent in both species (14.52% in camels and 3.03% in sheep). All cysts recovered from pigs were fertile. In Egypt, governmental agencies and veterinary authorities are asked to potentially eradicate stray dogs (the main definitive host) and towards the proper hygienic disposal of infected offal in abattoirs to minimize the prevalence of cystic echinococcosis., Competing Interests: Conflict of interestThe authors declare that they have no conflict of interests., (© Indian Society for Parasitology 2019.)

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 × 10 4 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 × 10 4 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.

Diclazuril, which is widely used for the prevention of coccidiosis in chickens, has a lethal effect on asexual and sexual stages of Eimeria spp. However, little is known about its effect on the exogenous stages of Eimeria spp. In this study, we evaluated the effect of in vitro treatment with 0.2% diclazuril on unsporulated and sporulated oocysts of Eimeria spp. For this purpose, a total of 180 male layer chicks aged one day were randomly divided into 5 experimental groups. Each group was divided into 3 replicates of 12 chicks each. Group 1 (G1) and Group 2 (G2) were negative (non-immunized and non-challenged) and positive (non-immunized and challenged) controls, respectively. Group 3 (G3) was immunized per os with 1.0 × 10 4 non-diclazuril treated-sporulated oocysts. Groups 4 (G4) was immunized per os with 0.2% diclazuril treated-unsporulated oocysts (1.0 × 10 4 ) in which diclazzuril didn't affect sporulation. Group 5 (G5) was immunized per os with 0.2% diclazuril treated-sporulated oocysts (1.0 × 10 4 ). Chicks of G2, G3, G4, and G5 were challenged with 7.5 × 10 4 untreated sporulated oocysts at the age of 21 days, while the group 1 chicks remained unchallenged. G4 and G5 animals immunized with 0.2% diclazuril-treated oocysts showed a significant decrease in bloody diarrhea severity, lesion scores, and oocyst counts in comparison to those immunized with untreated oocysts. Furthermore, histopathologic findings showed a low number of parasitic stages in cecal tissues in G4 and G5. A significant increased body weight gain was observed in Gs 4 and 5 in comparison to G2. In addition, expression levels of IL-2, IL-12, and IFN-γ were significantly increased in G4 and G5. In conclusion, diclazuril is effective in attenuating Eimeria oocysts and thus provides an alternative approach for using diclazuril-treated oocysts to protect chicks against Eimeria challenge., (Copyright © 2019 Elsevier B.V. All rights reserved.)

In the veterinary field, there is a great concern about the issue of acaricides resistance in cattle ticks. A comparative study was designed to evaluate the in vitro acaricidal efficacy of imidacloprid nano-emulsion, zinc oxide nanoparticles (ZnO NPs), silver nanoparticles (Ag NPs), deltamethrin, deltamethrin-ZnO NPs, and deltamethrin-Ag NPs. In addition, the novel formulations of deltamethrin (deltamethrin-ZnO NPs and deltamethrin-Ag NPs) against Rhipicephalus (Boophilus) annulatus were also evaluated by in vivo animal efficacy trials. Nanoparticles and their loaded forms, deltamethrin-Ag NPs and deltamethrin-ZnO NPs, were characterized by Transmission Electron Microscope (TEM) and Fourier-Transform Infrared Spectrum (FTIR). The adulticidal activity of deltamethrin-ZnO NPs at different concentrations; 2 mL/L, 1 mL/L, 0.5 mL/L and 0.25 mL/L induced a significant (P ≤ 0.05) lethal effect on adult ticks compared to deltamethrin-Ag NPs at the same concentrations. The larvicidal efficacy of deltamethrin-ZnO NPs resulted in a complete larval mortality within 24 h of exposure, while deltamethrin and deltamethrin-Ag NPs exhibited 100% immobility of larvae 48 h post-exposure. Furthermore, the in vivo experiments showed a considerable reduction in the tick's survival after using deltamethrin-ZnO NPs. There was no significant effect of different treatments on liver or kidney function tests at pre- and post-treatment of animals. In conclusion, deltamethrin-ZnO nanocomposite was the most effective adulticide and larvicide against R. (B.) annulatus. To the best of our knowledge, this is the first report for using deltamethrin-ZnO NPs as an acaricide., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The effect of six available and commercial disinfectants on the embryonation and larval development of Toxascaris leonina eggs was studied. Dettol® and Virkon® both induced a 100% reduction in larval development (P ≤ 0.05). Dettol® resulted in deformed eggshells and a halt in embryonal development at 1 week post exposure. All Virkon®-treated eggs showed an early embryonic lysis 24 h post exposure. TH4+ and 70% ethanol both significantly (P ≤ 0.05) affected larval development, with 58.8 and 85.8% reduction, respectively. Neither sodium hypochlorite nor phenol significantly affected larval development (2.8 and 21.0%, respectively). Sodium hypochlorite treatment caused a visible decortication of the eggshell; however, phenol-treated embryonated Toxascaris eggs appeared more or less morphologically normal. In conclusion, the disinfectants tested induced variable degrees of decortication and suppression of larval development. Virkon®S was the most effective disinfectant against Toxascaris eggs, suggesting that it is the most advisable one to use. To the best of our knowledge, this is the first report of the use of Virkon®S as an ovicide and/or larvicide of helminths, particularly Toxascaris leonina.

This study was designed to investigate the susceptibility/resistance of Rhipicephalus (Boophilus) annulatus to commercial deltamethrin 5%. The work was based on in vitro bioassays; adult immersion test (AIT) and larval packet test (LPT). Fully engorged female ticks were collected from 11 locations in Beni-Suef Governorate, Egypt (Alfashin, Eminelaros, Ehnasia, Smusta, Belifia, Aldiabia, Alhalabia, Beshnna, Alkom, Nasr and Beba). Two concentrations of commercial deltamethrin 5% (200 and 400 ppm) were tested by AIT on R. (B.) annulatus. The mortality of ticks at 400 ppm was higher than 200 ppm which was not reached to 100%. Ehnasia tick population was the least susceptible to deltamethrin while Alfashin population was the most susceptible one. Different deltamethrin concentrations (0.000625, 0.00125, 0.0025, 0.005, 0.010 and 0.020) were used in LPT to evaluate the larval mortality. The highest LC 50 appeared in Ehanasia tick population (0.01) with 95% confidence interval (0.005 to 0.01) meanwhile, the lowest LC 50 appeared in Alfashin (0.0006) with 95%CI (0.0005 to 0.0007). Thus, it is possible to deduce that one population was considered susceptible; seven populations were resistant level I and three populations were resistant level II. Additionally, PCR revealed that the resistant alleles (R) appeared in two localities (Ehnasia and Aldiabia). It was concluded that at least two tick populations of the investigated locations were of possible resistant to deltamethrin., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The activity of high doses of three insect growth regulators (IGRs), lufenuron (MATCH®), pyriproxfen® and hydroprene (Gentrol®), were tested on Rhipicephalus(Boophilus) annulatus adult females, eggs and larvae. Different concentrations of the IGRs were tested on eggs, larvae and adult ticks through immersion, larval packet and adult immersion bioassays, respectively. The tested IGRs did not show adulticidal activity against female ticks even at very high concentration. However, both hydroprene and pyriproxfen caused a significant decrease (P<0.05) in the reproductive indices of adult female ticks. Both lufenuron and pyriproxefen showed considerable ovicidal activity delaying the hatchability of the treated eggs until the 21st day and decreasing the hatchability percentages to 37.7% and 60.6% at concentrations ≥10X and ≥4X, respectively. Lufenuron (≥10X dose), hydroprene (≥4X dose) and pyriproxyfen (≥4X dose) induced highly significant larvicidal activity as they caused 100% mortality after 72 h of exposure. The oxidative profile of the hydroprene treated ticks had decreased glutathione peroxidase and increased malonaldehyde in comparison to the other IGR- treated and control untreated ticks. It is concluded that the IGRs did not show R. annulatus adulticidal effect, however, the deposited egg mass and its hatching percent decreased significantly when treated with hydroprene and pyriproxfen. The tested IGRs showed larvicidal activity against R. (B.) annulatus., (Copyright © 2018 Elsevier B.V. All rights reserved.)

This study was designed to investigate the ability of two herbal extracts and different chemical substances to inhibit or disrupt sporulation of Eimeria species oocysts of the chickens. The two herbal extracts were Allium sativum (garlic) and Moringa olifiera while the chemical substances included commercial disinfectants and diclazuril. Field isolates of Eimeria oocysts were propagated in chickens to obtain a continuous source of oocysts. The collected unsporulated oocysts (10 5 oocysts/5 ml) were dispensed into 5 cm Petri dish. Three replicates were used for each treatment. The treated oocysts were incubated for 48 h at 25-29 °C and 80% relative humidity. The results showed that herbal extracts, the commercial recommended dose of Dettol, TH4, Phenol, Virkon ® S, and Diclazuril 20% have no effect on the sporulation. While Sodium hypochlorite showed a significant degree of sporulation inhibition reached to 49.67%. Moreover, 70% ethanol, and 10% formalin showed 100% sporulation inhibition. It was concluded that 70% ethanol and 10% formalin are the most effective methods to inhibit Eimeria species sporulation., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Infection by Fasciola species was investigated in seven districts of Dakhla Oasis, Egypt, through abattoir inspection of cattle livers for adult worms and sedimentation of faecal samples from local cattle to detect Fasciola eggs. In addition, lymnaeid snails collected from the study area were examined microscopically for developmental stages of Fasciola spp. Abattoir inspection revealed that 51 out of 458 cattle livers (11.1%) contained adult flukes, which were identified morphologically as Fasciola hepatica. Examination of the cattle faecal samples revealed that 142 out of 503 (28.2%) contained Fasciola eggs. The collected snails, identified as Galba truncatula and Radix natalensis, showed larval stages of Fasciola in 71 out of 731 (9.7%) G. truncatula, while R. natalensis showed no infection. Specific duplex polymerase chain reaction (PCR) targeting the mitochondrial cox1 gene of F. hepatica and Fasciola gigantica was carried out on DNA extracted from pooled infected snails and adult worms. The F. hepatica size amplicon (1031 bp) was obtained from both the infected G. truncatula and the adult worms isolated from cattle livers from different districts. The amplicon sequences were identical to the published sequences of F. hepatica mitochondrial cox1 gene. In conclusion, the zoonotic importance of Fasciola infection and appropriate hygienic measures must be taken into consideration in Dakhla Oasis, Egypt.

Haemonchus contortus isolates were evaluated for benzimidazole (BZ) resistance or susceptibility by allele-specific PCR based on β-tubulin isotype 1 gene polymorphisms at the F167Y, E198A, and F200Y sites. Two isolates, one presumed susceptible from wild pronghorn antelope (PH) and one known to be resistant from goats (VM), were also assayed phenotypically for BZ resistance or susceptibility in the larval development assay (Drenchrite®). The BZ EC50 was 0.198 μM (intermediate between susceptible and weak resistant) for PH with critical well 5 (intermediate between susceptible and weak resistant) and 1.456 μM (intermediate weak resistant and resistant) for VM with critical well 8.5 (resistant). Genotypically, DNA extracted from pooled VM L3 larvae in the Drenchrite® wells with the highest BZ concentration was homozygous susceptible (SS) at the F167Y and E198A sites and homozygous resistant (RR) at the F200Y site by PCR, and sequence analysis bore this out. PH L3 larvae DNA from a control well (no BZ) was SS at all three sites by PCR, confirmed by sequence analysis. All single adult worm samples (N = 21) from PH, VM, Egypt goat (EG), and a Texas llama were SS at F167Y and E198A by PCR; however, only 3 PH worms and 1 EG worm were SS at F200Y. Three additional PH worms were RS and upon cloning two clones were identified as resistant by sequencing and two as susceptible. Clones from single adult worms VM, llama, and EG samples that were RR by PCR at F200Y were sequence verified as resistant. In this study, F200Y was the most frequently found genotypic marker for BZ resistance or susceptibility in the different Haemonchus isolates.

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., (Copyright © 2016 Elsevier Inc. All rights reserved.)

This study aimed to compare between the efficiency of in vivo fecal egg reduction test (FERT) and in vitro egg hatching assay (EHA) in evaluating of the anti-Fasciola activity of albendazole, triclabendazole, oxyclozanide and praziquantel. A field trial was carried out on fifty naturally Fasciola infected cattle that were divided equally into 5 groups (A-E). On day zero; groups A-D were drenched with albendazole, triclabendazole, oxyclozanide or praziquantel, respectively, while the remaining one, group E, was kept as untreated control. Fecal egg counts of the different groups were conducted weekly over a period of one month post-treatment. In vitro, commercial albendazole and oxyclozanide were diluted to 0.0002, 0.002, 0.02, 0.2 and 2.0 μg/ml, while commercial triclabendazole and praziquantel were diluted to concentrations of 25, 50, 75 and 100 μg/ml with dimethyl sulfoxide (DMSO). In vivo, at the 2nd week post-treatment, triclabendazole and oxyclozanide showed 100% fecal egg reduction (FER), and albendazole had a maximum of 73.7% reduction (P < 0.0001), however, praziquantel did not record any reduction of Fasciola egg counts. In vitro, triclabendazole treated Fasciola gigantica eggs showed early embryonic lysis with zero% hatching at the different concentrations (P < 0.01). In albendazole, the hatching varied according to the drug concentration. At the highest two concentrations; 0.2 and 2.0 μg/ml, the hatching percentages were 7.4 ± 1.6 and 5.6 ± 1.5 (P < 0.01) respectively. On the contrary, there were no significant differences in egg development and hatching percentage of oxyclozanide or praziquantel treated groups. In conclusion, the efficacy of triclabendazole and albendazole as fasciolicdes could be predicted by Egg Hatching Assay (EHA). Meanwhile fasciolicide activity of oxyclozanide could not be assessed with EHA. Based on in vivo and in vitro findings, paraziquantel did not show any fasciolicide effect., (Copyright © 2015 Elsevier B.V. All rights reserved.)