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.

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.

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.

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.

Synthesis of Carvacrol-Loaded Invasomes Nanoparticles Improved Acaricide Efficacy, Cuticle Invasion and Inhibition of Acetylcholinestrase against Hard Ticks.

Carvacrol is a monoterpenoid phenol found in many essential oils that has antibacterial, antifungal and antiparasitic activities. Drug loaded-invasome systems are used to deliver drugs utilizing nanoparticles to improve bioavailability, efficacy, and drug release duration. As a result, the present study developed carvacrol-loaded invasomes and evaluated their acaricidal effect against Rhipicephalus annulatus (cattle tick) and Rhipicephalus sanguineus (dog tick). Carvacrol loaded-invasome (CLI) was prepared and characterized using UV/Vis spectrophotometer, zeta potential measurements, Scanning Transmission Electron Microscopy (STEM), Fourier Transform Infrared (FT-IR) Spectroscopy, and Differential Scanning Calorimetry Analysis. CLI (5%) induced significant mortality (100%) in R. annulatus adult ticks with LC50 of 2.60%, whereas the LC50 of pure carvacrol was 4.30%. Carvacrol and CLI were shown to have a significant larvicidal action on both tick species, with LC50s of 0.24 and 0.21% against R. annulatus and 0.27 and 0.23% against R. sanguineus, respectively. Carvacrol and CLI (5%) induced significant repellent activities for 24 h against R. annulatus and R. sanguineus, as evidenced by the rod method and the petri-dish selective area choice method, respectively. High-performance liquid chromatography (HPLC) demonstrated that the CLI form had 3.86 times the permeability of pure carvacrol. Moreover, carvacrol and CLI inhibited acetylcholinesterase activity and decreased glutathione and malonedealdehyde levels in the treated ticks. In conclusion, invasomes significantly improved adulticidal and repellency activities of carvacrol against both tick species.

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.

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.

Potential of Marava arachidis, a Newly Recorded Earwig Species in Egypt as a Biological Control Agent of Rhipicephalus annulatus Tick in Laboratory.

In Egypt, only five species of Dermaptera (earwigs) have been reported. Based on both the morphological and molecular data of the earwig samples collected from a bakery in Beni-Suef, Egypt, we identified the species as Marava arachidis (Spongiphoridae), a cosmopolitan species with no prior records in Egypt. The current study was designed to analyze its predation capability on newly emerged eggs and larvae of the Rhipicephalus annulatus tick. A laboratory functional response study was set up by applying a predation test with various predator-prey ratios as treatments. This experiment was applied using the undefined mix of collected earwigs and the laboratory-collected eggs and the larvae of R. annulatus. The laboratory results showed that the mean number of predated tick eggs was 18.64 ± 11.18 in 24 h under the highest predator-prey ratio (1:10) examined, accompanied by 12.04 ± 4.38 broken but unconsumed eggs. Moreover, M. arachidis predated an average of 12.32 ± 9.07 tick larvae per day. In contrast, the mean dead larvae increased to 38.4 ± 2.30 per day with the highest predator density (1:10). The number of eggs and larvae consumed increased with the predator density. A linear relationship was detected between earwig density and the consumption rates of tick eggs (R2 = 0.99; p = 0.0001) and larvae (R2 = 0.96; p = 0.003). In conclusion, M. arachidis was first recorded in Egypt. This earwig has predation capability on R. annulatus tick eggs and larvae.

Remarks on Eimeria labbeana infection in domestic pigeons "Columbia livia domestica".

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 × 104 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.

Haemonchus contortus Susceptibility and Resistance to Anthelmintics in Naturally Infected Egyptian Sheep.

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 "ß-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.

Thymol efficacy against coccidiosis in pigeon (Columba livia domestica).

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 × 103sporulated 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.

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.

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.

Protective potential of diclazuril-treated oocysts against coccidiosis in layer chicks.

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 × 104 non-diclazuril treated-sporulated oocysts. Groups 4 (G4) was immunized per os with 0.2% diclazuril treated-unsporulated oocysts (1.0 × 104) in which diclazzuril didn't affect sporulation. Group 5 (G5) was immunized per os with 0.2% diclazuril treated-sporulated oocysts (1.0 × 104). Chicks of G2, G3, G4, and G5 were challenged with 7.5 × 104 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.

In vitro activity of natural and chemical products on sporulation of Eimeria species oocysts of chickens.

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 (105oocysts/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.

Molecular characterization of eimeria species naturally infecting egyptian baldi chickens.

BACKGROUND: Coccidiosis is a serious protozoal disease of poultry. The identification of Eimeria species has important implications for diagnosis and control as well as for epidemiology. The molecular characterization of Eimeria species infecting Egyptian baladi chickens was investigated. METHODS: Eimeria species oocysts were harvested from intestines of naturally infected Egyptian baldi chickens. The morphometry characterization of oocysts along with COCCIMORPH software was done. The DNA was extracted initially by freezing and thawing then the prepared samples was subjected to commercial DNA kits. The DNA products were analyzed through conventional polymerase chain reaction by using amplified region (SCAR) marker. RESULTS: The PCR results confirmed the presence of 7 Eimeria species in the examined fecal samples of Egyptian baldi breed with their specific ampilicon sizes being E. acervulina (811bp), E. brunette (626bp), E. tenella (539bp), E. maxima (272bp), E. necatrix (200bp), E. mitis (327bp) and E. praecopx (354bp). A sequencing of the two most predominant species of Eimeria was done, on E. tenella and E. máxima. Analysis of the obtained sequences revealed high identities 99% between Egyptian isolates and the reference one. Similarly, E. maxima isolated from Egyptian baldi chickens showed 98% nucleotide identities with the reference strain. Only single nucleotide substitution was observed among the Egyptian E. tenella isolates (A181G) when compared to the reference one. The Egyptian isolates acquired 4 unique mutations (A68T, C164T, G190A and C227G) in compared with the reference sequence. CONCLUSION: This is the first time to identify the 7 species of Eimeria from Egyptian baladi chickens.