Acta Parasitologica

Morphological and Molecular Characteristics of the Gastro-Intestinal Nematode Parasite Ascaridia columbae Infecting the Domestic Pigeon Columba livia domestica in Saudi Arabia Abstract Background Parasitism is a complex problem that is often ignored in companion animals, including birds, unless it develops into a severe clinical disorder. The present study was, therefore, aimed to investigate the presence of the gastrointestinal nematode infecting the domestic pigeon and provide a complete morphological description and clarify its taxonomic position through phylogenetic analysis of the ITS1-5.8s-ITS2 rDNA gene region. Materials and Methods During the current study, a total of twenty-six domestic pigeons, Columba livia domestica, were collected and internal organs examined for helminth detection. Using light and scanning electron microscopy, the recovered parasite species are studied. In addition, the selected gene region was obtained and sequenced using appropriate primers that aid in the formation of the phylogenetic dendrogram for the recovered parasite species with others retrieved from GenBank. Results Morphological examination showed that this nematode parasite belongs to the Ascaridiidae family within the genus Ascaridia. The material was assigned to the previously described Ascaridia columbae by providing all the characteristic features as the presence of a mouth opening surrounded by three tri-lobed lips; each lip has two triangular teeth with a spoon-like structure, cephalic papillae and amphidal pores on lips surface, presence of lateral cuticular alae and pre-cloacal sucker, 10 pairs of caudal papillae, and two equal spicules in male worms. The morphological investigations of this species were supplemented by molecular analysis of ITS1-5.8s-ITS2 rDNA gene region. The data showed that the present A. coulmbae is deeply embedded in the Ascaridia genus with a 74–99% sequence similarity to other species in the Chromadorea class. Ascaridiidae appears as monophyly and represented as a sister group to Heterakidae. The ascaridiid species examined belong to the Ascaridia genus and displaced a close relationship with the previously described A. coulmbae (gb| KF147909.1, gb| AJ001509.1, gb| KC905082.1, gb| JQ995321.1, gb| JX624729.1) as putative sister taxa. Conclusion The present study revealed that the species Ascaridia is the first account of this genus as an endoparasite from the domestic pigeon inhabiting Saudi Arabia. Therefore, the combination of morphological and molecular studies helps to identify this species correctly and identified as Ascaridia columbae.

In vitro Assessment of Camphor Hydrazone Derivatives as an Agent Against Leishmania amazonensis Abstract Purpose Due to serious problems with the treatment of leishmaniasis all around the world, here is an urgent need in the search for new drugs that are more effective and safer for the treatment of the various forms of leishmaniasis. Actual therapy is limited and lacks sufficient efficacy due to incomplete elimination of the parasites form of patients. In this sense, we decided to evaluate, by first-time, a series of seventeen camphor hydrazone derivatives (2a–2p) against Leishmania amazonensis. Methods The compounds previously synthesized from camphor, an abundant natural compound, were evaluated in vitro against the extra and intracellular forms of Leishmania amazonensis, and murine macrophages. Results The majority of compounds, fourteen, displayed activity against the intracellular form of the parasite (amastigote) with IC50 values ranging from 21.78 to 58.23 µM, being six compounds active for both forms of the parasite. The compound 2i exhibited higher activity against the amastigote form with the value of IC50 (21.78 µM) close to standard utilized miltefosine (12.74 µM) and selectivity index of at least 6.9. Six compounds displayed activity against promastigote form of Leishmania amazonensis 2g, 2j–2n (41.17–69.59 µM), with the compound 2m being the more active with IC50 = 41.17 µM, 1.9 times less active than the reference drug (IC50 = 21.39 µM). The compound 2m was the more selective to this form, with a selectivity index of at least 3.6. All the compounds were non-cytotoxic to macrophages. Conclusions Most compounds showed activity against amastigote form of Leishmania amazonensis, being that they were not cytotoxic to macrophage at the maximum tested concentration, showing the selective property of these compounds. Since amastigotes are the parasite stages that cause the disease in humans, these results highlight the antileishmanial effect of the compounds. This study indicates the possible development of candidates to leishmanicidal drugs from an abundant natural compound of easy access.

The genomic survey of Tc1-like elements in the silkworm microsporidia Nosema bombycis Abstract Background Microsporidia Nosema bombycis is the destructive pathogen in the production of sericulture. The Tc1/mariner elements belong to important component of DNA transposon. Methods The genomic data of N. bombycis and related Nosema species were screened to identify the Tc1-like elements and analyzed the phylogenetic relationship, based on bioinformational analysis. High-throughput data of transcriptomes and small RNAs were used to evaluate the expressed level and potential rasiRNAs for the Tc1-like elements of N. bombycis. Results Twelve complete Tc1-like elements belonging to DD34,E clade is confirmed in the whole genome of N. bombycis, and divided into two branches. Six of them are sole in N. bombycis and thereby would be the molecular marker to differentiate this species from others Nosema spp. Most of the elements have the transcriptional active and are the source of sRNAs. Conclusion Abundant Tc1-like elements in N. bombycis reflect the expansion of transposons for this genomic characters, comparing with others Nosema spp. The finding of distribution, phylogeny and potential functional activity for Tc1Nbs in N. bombycis will help understanding the role of the DNA transposon in genomic evolution of microsporidia.

Molecular Diagnosis and Pathological Study of Toxoplasma gondii in Aborted Caprine and Ovine Fetuses in Borderline of Iran–Iraq Abstract Background Infection with the protozoan parasite Toxoplasma gondii is widely prevalent in animals and humans. In goats and sheep this infection has a high economic impact as it causes abortions and lamb losses. Although there are several studies reporting seroprevalence in small ruminants in Iran, molecular-based information is scarce. Purpose This study aimed to screen caprine and ovine aborted fetuses for T. gondii infection by PCR and histopathology. Methods Brains of 121 aborted fetuses (10 caprine and 111 ovine) were collected from different parts of the Kordestan province, bordering with Iraq. Gestational age and the general status of each fetus such as freshness, autolysis, mummification and presence of macroscopic lesions in the fetus and foetal membranes was recorded. Individual brain tissues of fetuses were subjected to nested-PCR targeting the B1 gene, and histopathological sections prepared from brains were examined microscopically. Results PCR results revealed T. gondii-associated abortion in one caprine and nine ovine fetuses (8.3%). Microscopically, pathological lesions included non-purulent meningitis associated with gliosis, focal necrosis, and occasionally tissue cyst. Conclusion This paper reports for the first time T. gondii-associated abortion in goats in Iran. As organs from aborted fetuses do not necessarily show lesions, molecular confirmation is the unique diagnostic method and should be used in situations of an abortion epidemic.

Genotyping by Sequencing of Acanthamoeba and Naegleria Isolates from the Thermal Pool Distributed Throughout Turkey Abstract Purpose The main goal of this study was genotyping of free-living parasites and sub-grouping of pathogenic or non-pathogenic amebae obtained from Turkey’s thermal springs. In so doing, distribution and abundance of possible pathogenic or causative strain for humans, which are caused by Acanthamoeba and Naegleria strains, would be elaborated. The number of extensive studies on the general occurrence and distribution of parasitic strains is very high worldwide, but there has been a paucity of information with regard to Turkey. Methods From a total of 434 obtained thermal pool samples, free-living amebas were isolated from 148 water samples using the non-nutrient agar (NNA) culture method. Subsequently, the cultivated samples were used for DNA isolation; then 102 obtained DNA samples were subjected to PCR amplification using various primers for samples of genera Acanthamoeba and Naegleria. Ultimately, estimation of genotype or subtype was evaluated by sequencing. Results About 29 samples that belong to Acanthamoeba and Naegleria were estimated from a total of 102 amplified PCR samples. These eukaryotic PCR products which have Acanthamoeba genus appearance, generated 26 subtypes and 3 Naegleria samples. Among the 26 Acanthamoeba genotypes, 22 aligned sequences were matched with various GenBank reference samples, while the 4 divergent genotypes were not elaborated and marked as ND. Most of the Acanthamoeba genera were determined as likely dominating groups and clustered as T form within totally eight groups. Eight, seven and three subtypes were found as T4A, T15 and T11 genotypes, respectively while the remainings were ultimately found in four groups. Results confirming the predominance of T4A, which is known the most causative form, the presence in the pools. Despite being uncommon, N. fowleri, lovaniensis and australiensis were also observed among the surveyed pools. Conclusion The present study is descriptive and is not unique. However, this is the most comprehensive study of the molecular distribution sampling of thermophilic Acanthamoeba and Naegleria that confirmed and demonstrated their ubiquitous presence throughout Turkey. By this estimation, in some spas, the most and likely causative form Acanthamoeba including T4 and Naegleria fowleri has also been confirmed.

Molecular Prevalence and Epidemiology of Trypanosoma evansi Among Cattle in Peninsular Malaysia Abstract Background Animal trypanosomiasis (Surra) caused by Trypanosoma evansi (T. evansi) is known to be one of the important haemoprotozoan parasites that causes great economical loss on animal production due to mortality and loss of condition. Methods A cross-sectional study was designed to evaluate the prevalence and risk factors associated with T. evansi infection among cattle in Peninsular Malaysia. Polymerase chain reaction (PCR) was employed on 1045 blood samples collected from 43 farms. A well-structured questionnaire was used to collect data on risk factors associated with T. evansi prevalence. The RoTat 1.2 set of primers was used to amplify products of 205 base pair. Results The overall prevalence was found to be 17.9% (187/1045; 95% CI = 15.66–20.31). Trypanosoma evansi was detected among cattle in all the States of Peninsular Malaysia. Breeds of cattle and closeness to waste area, where the risk factors significantly (p < 0.05) associated with the PCR positivity of T. evansi among cattle in Peninsular Malaysia. Conclusion This appears to be the first time a comprehensive survey on the prevalence and risk factors of T. evansi infection in cattle using molecular tools is been carried out in the entire states of Peninsular Malaysia. The findings from this study will provide baseline information on the molecular prevalence of the haemoflagellate and its associated risk factors among cattle for an improved beef and dairy production in Peninsular Malaysia.

Effectiveness of Repeated Administration of Praziquantel with Disodium Glycyrrhizinate and Two Enantiomers of Praziquantel on Opisthorchis felineus (Rivolta, 1884) Abstract Background Nowadays, it is still important to develop effective anti-opisthorchiasis agents. In this work, we tested a complex of praziquantel (PZQ) with a plant origin compound—disodium glycyrrhizinate—in the ratio 1:10 PZQ:Na2GA, containing 11-fold less of the active ingredient. Our aim was to study various ways to treat trematode Opisthorchis felineus with this complex in vitro. Additionally, an in vitro comparison of the anthelmintic action was made among racemic-PZQ, (R)-PZQ, and (S)-PZQ on juvenile and adult maritae of O. felineus. Methods Worms extracted from the hamsters were subjected to various regimens of administration of the complex: once a day for 3 days or three times within 1 day. Moreover, mature maritae and juvenile worms of O. felineus were subjected to the comparison the anthelmintic effectiveness of racemic-PZQ, (R)-PZQ, and (S)-PZQ. Results The O. felineus maritae that received PZQ:Na2GA (1:10) thrice within 1 day were most strongly affected by the drug. Their motility substantially decreased already on the second day after the last dose, and the percentage of live worms by the end of the experimental period was the lowest. These results indicate a cumulative anthelmintic effect of this substance under the regimen “three times within 1 day.” For the first time, we report that among the three substances (racemic-PZQ and two enantiomers), (R)-PZQ has the highest anthelmintic activity, toward both juvenile and sexually mature maritae of O. felineus. Conclusion These findings suggest that the development of a supramolecular complex of (R)-PZQ with disodium glycyrrhizinate and administration of this complex three times within 1 day are promising approaches.

Correction to: Saving the Manx Shearwater Flea Ceratophyllus ( Emmareus ) fionnus (Insecta: Siphonaptera): The Road to Developing a Recovery Plan for a Threatened Ectoparasite Due to a processing error, the presentation of Table 1 was incorrect. The corrected table is given below. The original article has been corrected.

Correction to: Persistence of High In Vivo Efficacy and Safety of Artesunate–Amodiaquine and Artemether–Lumefantrine as the First- and Second-Line Treatments for Uncomplicated Plasmodium falciparum Malaria 10 Years After Their Implementation in Gabon Unfortunately two errors appeared in this article.

Retrospective Study of Cystic Echinococcosis (CE) Based on Hospital Record from Five Major Metropolitan Cities of Pakistan Abstract Background Human cystic echinococcosis (CE) is an important zoonotic parasitic disease. It is one of the NTDs having strong roots in Pakistan. There are limited studies on the prevalence of CE in Pakistan. Method A retrospective study was conducted to determine the geographical distribution of human CE in Punjab (Lahore, Multan, and Faisalabad) and Sindh (Karachi and Hyderabad) provinces of Pakistan. Clinical records of infected patients from nine major hospitals were retrieved from 2008 to 2018. Results A total of 188 surgically confirmed cases were included from selected hospitals during the study period, which showed the annual frequency of 18.8 cases/year. The number of reported cases was higher in Sindh province (67.55%) compared to Punjab (32.45%). Total number of CE cases/city was higher in Karachi (47.34%) compared to Hyderabad city (20.21%), while in Punjab province, this frequency was higher in Lahore (25%) followed by Multan (5.32%) and Faisalabad (2.13%). Out of total 188 patients, 73 (38.83%) were males and 115 (61.7%) were females. In surveyed hospitals, high burden of infection was reported in young adults (21–30 years; 29.79%) followed by 21.81% and 17.02% in 31–40 and 41–50 age groups, respectively. Data retrieved from the hospital record of diagnosed cases of CE showed liver (36.17%, n = 68) as the most affected organ followed by lungs (26.06%, n = 49). In this survey, some unusual sites of cyst formation were also observed. Conclusion Current study is a step forward towards filling the gap of knowledge for prevalence and regional distribution of hydatidosis in Pakistan reporting the CE cases in humans for the first time in Multan and Faisalabad.