Molecular characterization of the unique Mesostephanus appendiculatus (Trematoda: Cyathocotylidae) by small ribosomal RNA from Egypt.

M esostephanus appendiculatus (Family: Cyathocotylidae) is one of the unique trematodes that complete their cycles in human and animal intestines in many countries of the world. The main source of its transmission is eating raw or undercooked infected fish muscle. Earliest analyses of genes to different parasites supported the analysis of helminthes either biological or morphological. This paper detected M. appendiculatus sequence with GenBank accession number gb (KY026782). Comparison of M. appendiculatus with other helminthes using BioEdit 7 and MEGA7 program shows some similarity in different points along its sequence. The phylogenetic analysis clarifies that it was closely related to both trematodes (Clinostomum complanatum and Echinochasmus japonicus) and some cestodes of fish origin such as Polyonchobothrium polypteri, Bothriocephalus sp., and Haplobothrium globuliforme. The obtained results provide a good source for genome analysis of M. appendiculatus in relation to other Platyhelminthes.

New approach to molecular characterization of Paramphistomum cervi and Carmyerius gregarius and comparative analyses with selected trematodes.

The two ruminant parasites, Paramphistomum cervi and Carmyerius gregarius, were collected from fresh-slaughtered native cattle at local abattoirs in Sadat district, Menoufia province and identified morphologically, then molecularly by sequencing the nucleotides of 18S ribosomal RNA gene (18S rRNA). The nucleotide sequences of the two isolates were 456 (P. cervi) and 401 bases for (C. gregarius). The data were used along with those of several other helminth species from the GenBank to identify these two species genetically. The nucleotide sequences were aligned using multiple sequence alignments of nucleotides by Clustal W 12.1 V and construct their relationship. Neighbor-joining analytical method was used showing sister relationship between C. gregarius from Sadat district and Gastrodiscoides hominis (EF027096) with relative identity of (98%) due to the presence of single nucleotides polymorphisms (SNPs) in the form of indels as nine nucleotides positions. But when clustering of P. cervi Sadat isolate with Paramphistomoidea sp. S4 isolate P5 (GU735643), this relationship shows complete identity (99%) between them. The homology and diversity was done using Bayesian analyses in MrBayes v3.1. This work will give a useful guide for other researchers for the molecular taxonomic position of Paramphistomatidae spp. in Sadat district among the different species around the world.

Efficacy of deltamethrin, diazinon, and ivermectin on Boophilus annulatus ticks (in vitro and in vivo study).

Tick infection is considered a cause of major concern as it is a vector for some disease transmission. The use of chemicals to control tick infection is increasing in farm systems. The efficacy of three chemicals was studied on the tick Boophilus annulatus. In vitro and in vivo studies were done. The active ticks were collected from naturally infected cattle for in vitro study. They were incubated with the three chemicals which are commercially used. An in vitro study recorded that the highest effect of the three chemicals was 100% at 3 h postexposure (p.e.) time for deltamethrin and 6 h for diazinon and ivermectin on the adult ticks. Egg batches were less affected. In vivo results showed more plain efficacy. The efficacy of deltamethrin was increased gradually until complete cessation of ticks showed within 3rd day posttreatment (d.p.t.), 100% efficacy. But the tick population begins to reappear gradually within 7 d.p.t., while diazinon showed 100% efficacy at 7 d.p.t. and the ticks reappear again within 14 d.p.t. The most preferred results were obtained with ivermectin which showed 100% efficacy at 7 d.p.t., and the cattle was still free from infection until 21 d.p.t. only. Ticks begin to reappear within 28 d.p.t. in slight few numbers. This concluded that the powerful and safe chemical which is commercially used was ivermectin. Even so, it is used also as an anthelmintic drug.

Anthelmintic activity of ginger, curcumin, and praziquentel against Raillietina cesticillus (in vitro and in vivo).

This work evaluates the anthelminitic activity of ginger and curcumin on the cestode Raillietina cesticillus. Live parasites were collected from intestine of naturally infected chickens in PBS 0.9 % and then incubated at 37 °C in media containing ginger extract at three different concentrations (125, 250, and 500 mg); every concentration was dissolved in 10 ml media. The curcumin extract was used at three different concentrations (250, 500, and 1000 mg); each was dissolved in 10 ml media. Praziquantel at a concentration of 600 mg was added to 10 ml media. A control one without extract was reported. Regression of worms increased gradually in all concentrations. At 500 mg ginger (50 ± 0 %), worms were regressed at 48 h post-exposure (h.p.e.). Also (50 ± 1.8 %), worms were regressed at 1000 mg curcumin at the same time. On the other hand, praziquantel showed the highest regression (65 ± 2.3 %). The extract efficacy was exhibit as concentration-time-dependent mainly at higher concentrations used after 48 h. In vivo effects of ginger and curcumin were lower than those in vitro.

Acaricidal Efficacy of Thirty-Five Egyptian Plants Against the Camel Tick, Hyalomma Dromedarii.

PURPOSE: Alternative and affordable tick control strategies are crucial to control and prevent tick bites and tick-borne diseases. METHODS: In this study, we evaluated the acaricidal efficacy of 35 aqueous plant extracts (17%) against the camel tick, Hyalomma dromedarii. RESULTS: The phytochemical profile indicated the presence of various secondary substances. Plants were classified into three groups according to their mortality percentage 15 days post-treatment with 17%. This highly effective group (91%-95%) comprised Ocimum basilicum, Mespilus germanica, and Viola alpine followed by Carum carvi, Cucurbita pepo (peel), and Peganum harmala. A moderately effective group (80%-90%) included Acacia nilotica, Apium graveolens, Capsicum annuum, Ceratonia siliqua, Cucurbita pepo (seeds), Equisetum arvense, Eruca sativa, Ginkgo biloba, Plantago psyllium, Phyllanthus emblica, Punica granatum, and Ziziphus spinachristi. The 20 remaining plants were assigned to the less effective group (< 80%). Viscum album (58.3%), which was the least effective reference plant. The high potency of six plant extracts as acaricides may be attributed to the high content of active principles, e.g., phenols, flavonoids, and tannins. CONCLUSION: All of these highly effective plants are recommended for use as an acaricide, in case of facing acaricidal resistance or limited options for tick control.

Molecular characterization of Ascaridia galli infecting native chickens in Egypt.

Family: Ascaridae as a whole is distributed among Africa and adjacent regions and in many areas of the world. The nematode Ascaridia galli is one of the most pathogenic and economically important parasites of poultry. The adult affect the small intestine of the hosts feeding on digested food materials. Its control costs million dollars annually. The genomic DNA was extracted from nematode parasites, A. galli, from specific host, native chickens. The polymerase chain reaction (PCR) was applied to ensure that the DNA content aids in the further studies. Two primers were used in the PCR reactions. The two primers were screened, only the second primer gave total amplified fragment markers 818 bp. The gene sequences obtained from Egyptian A. galli was compared with another one of accession number (AY587609) showing that the sequence was similar in some points from 346 to 1244 sequence, to make a phylogenetic relationships of A. galli with other nematodes on the data base showing that it was to some extent similar to Heterorhabditis spp.

Molecular identification and phylogenetic analysis of Heterakis gallinae from native chickens in Egypt.

The genomic DNA was extracted from nematode parasites, Heterakis gallinae from specific host, native chickens. The polymerase chain reaction (PCR) was applied to make amplification of the DNA content that aids in the further studies. The primer was used in the PCR reactions that have the following sequences: forward (5'-GTTTCCGTAGGTGAACCTGC-3') and reverse (5'-ATATGCTTAAGTTCAGCGGGT-3'). It gave total amplified fragment markers at 914 bp. The gene sequences obtained from Egyptian H. gallinae was to make phylogenetic relationships of Egyptian H. gallinae with other nematodes on the database of an accession number (AJ876757), showing that it was nearly similar to Heterakis gallinarum.

In vitro and in vivo screening of anthelmintic activity of ginger and curcumin on Ascaridia galli.

Intestinal helmintic infection, continue to be a cause of major concern in several parts of the world, particularly in the developing nations. The use of plant extracts to control poultry helminths is increasing in different rearing systems. The anthelmintic activity of ginger and curcumin was studied on the nematode Ascaridia galli. In vitro and in vivo studies were allocated. Live parasites for in vitro studies were collected from the intestine of naturally infected chickens. Some living worms were incubated at 37 °C in media containing ginger at three concentration levels (25, 50, and 100 mg/ml), and others were incubated in media containing curcumin at the same concentration levels. Another living worm group was incubated in media containing albendazole at a dose of 7.5 mg/ml. The extracts' efficacy was exhibited in a concentration-time-dependent manner mainly at 100 mg/ml and after 48 h. The in vivo study takes place on experimentally infected chickens. Group of infected chickens was treated with ginger extract at dose of 100 mg, another group was treated with curcumin extract at dose of 100 mg, and a third group was treated with albendazole at dose of 7.5 mg. In vivo study of ginger and curcumin recorded lower mortality rates than the in vitro study. It is concluded that ginger and curcumin extracts have potential anthelmintic properties against A. galli. Ginger in all concentrations used exhibited a higher death rate observed than curcumin. Their wormicidal effect is concentration-time dependent.

Pathological and genetic characterization of foot and mouth disease viruses collected from cattle and water buffalo in Egypt.

Foot-and-mouth disease (FMD), a highly contagious viral disease caused by FMD virus (FMDV) that threatens Egypt's livestock industry. FMDV causes severe economic losses in the livestock, with restriction of international trade from endemic regions. Surveillance for FMDV serotypes circulating in Egypt is urgently needed to assess the epidemiological situation in the country. FMD outbreaks reported in Egypt in between December 2016 and January-March 2017. A cross-sectional study was conducted to identify the FMDV serotypes responsible for the outbreaks and to collect information on the virus's morphopathological effects. Postmortem tissue and clinical samples (oral swabs, vesicular fluids from ruptured vesicles, and blood) were collected from recently deceased and infected animals. Pathological examination revealed classical FMD lesions as vesicular and erosive lesions on epithelial tissues with non-suppurative lymphoplasmacytic myocarditis. Phylogenetic and sequencing analyses demonstrated that FMDV serotype O, EA-3 topotype, VP1 is the prevalent serotype responsible for the pathological alterations and the high mortality in young calves, adult cattle, and water buffalo. The outcomes indicate continuous mutations in the circulating FMDV, which result in the occasional failure of vaccination. Based on these findings, extensive continuous monitoring and serotyping of the existing circulating FMDV isolates and regular vaccination with reevaluation of the currently used vaccine in Egypt are recommended to prevent the recurrence of such outbreaks.

ELISA and some biochemical tests of heterophyidae infection in laboratory animals.

Heterophyiasis is an important food-borne parasitic zoonosis in Egypt, among the inhabitants living around brackish-water lakes especially fishermen, and it is a common human parasite in the Nile Delta. The experiment was done on two laboratory animals (rats and dogs), and the time of sample collection was done periodically at 6, 9, 15, 21, and 28 days post-infection to evaluate different tests required. Whole blood was collected with heparin or ethylenediamine tetra-acetic acid as anticoagulant to help in the hematological studies such as red blood cells count (RBCs), white blood cells count, packed cell volume (PCV), and hemoglobin (Hb). Only marked increase in the total leuckocytic count was recorded while RBCs, PCV, and Hb were decreased in most of the results obtained. Total protein and globulin decreased while albumin and A/G ratio increased. Liver enzymes showing marked increase in aspartate aminotransferase and increase in alanine aminotransferase in dogs and rats denoting that liver has a role in the response to that infection. Kidney-function tests, urea, and creatinine showed slight increase at 6 days post-infection (d.p.i.). After preparation of different Ag (antigen) from different collected helminthes, the protein content of each was determined. The sera of infected animals were collected to find antibodies in their blood against the parasite using enzyme-linked immunosorbent assay and using crude heterophyid antigen collected from their intestines after scarification. The worms washed, homogenized, and then centrifuged to collect supernatant fluid as antigens. The results indicated that antibody starts to appear at 9 d.p.i. and increases till 21 and 28 d.p.i. and detection depends on antigen concentration.

Onchocerca flexuosa. sp. (Nematoda: Filarioidea) in Japanese Wild Sika Deer (Cervus nippon): Pathological and molecular identification.

Subcutaneous nodular onchocercosis was detected and investigated in 17 Japanese Sika deer (Cervus nippon), captured in Gifu and Shiga Prefectures/Japan, in the period between 2016 and 2017. The worms were found in all the seventeen deer within characteristics subcutaneous nodules dispersed mainly in the back (especially in the lumbar region and flanks), with few scattered nodules were located at the forelimbs and neck. The all collected nodules were examined stereo-microscopically. The parasites were extracted from the nodules and identified through morphological and histopathological examinations. Molecular identification through sequencing of the following genes; internal transcribed spacer subunit 2 (ITS2)-28S ribosomal RNA (28S rRNA), cytochrome c oxidase subunit 1 (cox1) and mitochondrially encoded NADH dehydrogenase subunit 2 (NAD2) were performed. The histopathological, molecular and phylogenetic analysis demonstrated that, the filarial nematode isolated from Gifu and Shiga Prefectures in Japan is O. flexuosa. This is the first report about presence of O. flexuosa in Japanese Sika deer (Cervus nippon) in Gifu and Shiga Prefectures. Supplementary Information: The online version supplementary material available at 10.1007/s12639-021-01453-3.

First report of Posthodiplostomum minimum metacercariae and resulting histopathological changes in Bagrus bajad from Lake Nasser, Egypt.

Parasitological studies of 100 Bagrus bajad fish were collected from Lake Nasser, Aswan, Egypt. Out of them 75 were infected with metacercariae in two forms; encysted in most visceral organs and free only in air bladder. These were identified morphologically as Posthodiplostomum minimum metacercariae (Strigeidida: Diplostomidae). The intensity of white grub in B. bajad was high in the intestine (30 encysted metacercariae/g). The highest prevalence season observed in summer and the lowest in winter. The histopathological examination revealed encysted metacercariae of different sizes within the intestinal musculature accompanied with granular infiltration with eosinophilia. Degenerative changes were observed in liver and kidney cells, as well as congestion of hepatic sinusoids. Spleen showed both lymphoid and melanomacrophages cells depletion.

Epidemiology of bovine fascioliasis in the Nile Delta region of Egypt: Its prevalence, evaluation of risk factors, and its economic significance.

AIM: This study focuses on the risk factors associated with the prevalence of Fasciola affecting cattle population in three provinces belonging to the Nile Delta of Egypt and to estimate the economic losses as a result of fascioliasis. MATERIALS AND METHODS: From January 2015 to end of December 2015, records of 21 farms (4976 cattle) were analyzed to screen the prevalence of fascioliasis among cattle farms, to identify its associated risk factors and its economic impacts on Nile Delta region of Egypt. RESULTS: The overall prevalence of fascioliasis in the Nile Delta region of Egypt was 9.77%. The prevalence of fascioliasis was found to be statistically significantly associated with age, sex, breed, and type of farms. The highest prevalence was observed in <2 age group (10.91%), and the lowest was >3 age groups (8.35%). In terms of body condition scores, cattle with medium and poor conditions were associated with fascioliasis more than those with good body condition. Besides, cattle raised in organic farms were associated with lower risk of fascioliasis than those in conventional farms. The prevalence of fascioliasis was noted more prominent in districts with moderate temperatures and with relative humidity (>60%). The annual overall costs for fascioliasis were estimated to be 221.2 USD/cow due to the significant reduction in body weight, reduction in milk production, and the treatment costs for fascioliasis. CONCLUSION: The results provided could be helpful for improving the control and preventive strategies.