In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts.

Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

In-vitro evaluation of Nigella sativa and Punica granatum effect on protoscolices of hydatid cysts / Avaliação in vitro do efeito de Nigella sativa e Punica granatum em protoescólices de cistos hidáticos

Abstract Cystic echinococcosis (CE) are commonly found in the liver and lungs of affected hosts. The treatment approach is usually surgical, or giving drugs in conjunction before surgery to kill protoscolices, to avoid anaphylactic shock from leakage of hydatid fluid into the peritoneum and to decrease opportunities for recurrences. The present study was to evaluate the in vitro scolicidal efficacy of hydroalcoholic extract of Punica granatum peel and Nigella sativa, on the protoscolices of CE that collected from the lungs of infected camels. Different concentrations of extracts with different exposure times were used and a viability assay was applied to measure the scolicidal effect. N. sativa showed its highest scolicidal efficacy at 100 mg/mL and 10 mg/mL concentrations after 30 and 60 min. P. granatum peel extract showed its maximum scolicidal efficacy at 100 mg/mL concentration after 120 min. All experiments of the current study revealed that the extracts of both N. sativa and P. granatum had a scolicidal effects on the protoscolices of camel hydatid cysts. It could be concluded that N. sativa extract is more potent than P. granatum peel extract regarding scolicidal effect, but the efficacies of both extracts were of moderate significant correlation to exposure time and concentrations.

Resumo Os cistos hidáticos (equinococose cística, CE) são comumente encontrados no fígado e nos pulmões dos hospedeiros afetados. A abordagem do tratamento geralmente é cirúrgica, e algumas drogas são administradas em conjunto antes da cirurgia para matar protoscólices e evitar choque anafilático devido ao vazamento de fluido hidático no peritônio e diminuir as oportunidades de recorrência. O presente estudo foi avaliar a eficácia in-vitro do extrato hidroalcoólico de casca de Punica granatum e Nigella sativa, sobre os protoescólices de cistos hidáticos, que foram coletados dos pulmões de camelos infectados. Concentrações dos extratos com diferentes tempos de exposição foram utilizados e um ensaio de viabilidade foi aplicado para medir o efeito escolicida. A N. sativa apresentou sua maior eficácia escolicida nas concentrações de 100 mg/mL e 10 mg/mL após 30 e 60 min. O extrato de casca de P. granatum mostrou sua máxima eficácia escolicida na concentração de 100 mg/mL após 120 min. Todos os experimentos do presente estudo revelaram que os extratos de N. sativa e P. granatum tiveram efeito escolicida dependente da dose e do tempo nos protoescólices dos cistos hidáticos de camelo. Pode-se concluir que o extrato de N. sativa é mais potente que o extrato de casca de P. granatum em relação ao efeito escolicida, mas a eficácia de ambos os extratos foi de correlação significativa moderada com o tempo de exposição e as concentrações.

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.

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.

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.

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.

Apicoplast-targeting antibacterials inhibit the growth of Babesia parasites.

The apicoplast housekeeping machinery, specifically apicoplast DNA replication, transcription, and translation, was targeted by ciprofloxacin, thiostrepton, and rifampin, respectively, in the in vitro cultures of four Babesia species. Furthermore, the in vivo effect of thiostrepton on the growth cycle of Babesia microti in BALB/c mice was evaluated. The drugs caused significant inhibition of growth from an initial parasitemia of 1% for Babesia bovis, with 50% inhibitory concentrations (IC(50)s) of 8.3, 11.5, 12, and 126.6 µM for ciprofloxacin, thiostrepton, rifampin, and clindamycin, respectively. The IC(50)s for the inhibition of Babesia bigemina growth were 15.8 µM for ciprofloxacin, 8.2 µM for thiostrepton, 8.3 µM for rifampin, and 206 µM for clindamycin. The IC(50)s for Babesia caballi were 2.7 µM for ciprofloxacin, 2.7 µM for thiostrepton, 4.7 µM for rifampin, and 4.7 µM for clindamycin. The IC(50)s for the inhibition of Babesia equi growth were 2.5 µM for ciprofloxacin, 6.4 µM for thiostrepton, 4.1 µM for rifampin, and 27.2 µM for clindamycin. Furthermore, an inhibitory effect was revealed for cultures with an initial parasitemia of either 10 or 7% for Babesia bovis or Babesia bigemina, respectively. The three inhibitors caused immediate death of Babesia bovis and Babesia equi. The inhibitory effects of ciprofloxacin, thiostrepton, and rifampin were confirmed by reverse transcription-PCR. Thiostrepton at a dose of 500 mg/kg of body weight resulted in 77.5% inhibition of Babesia microti growth in BALB/c mice. These results implicate the apicoplast as a potential chemotherapeutic target for babesiosis.

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.