Published by De Gruyter

Volume 63 Issue 2

Issue of Acta Parasitologica

Contents
Journal Overview

April 13, 2018

Morphological, histological and molecular characterization of Myxobolus kingchowensis and Thelohanellus cf. sinensis infecting gibel carp Carassius auratus gibelio (Bloch, 1782)

Bo Zhang, Yanhua Zhai, Zemao Gu, Yang Liu

Page range: 221-231

Abstract

A Myxobolus species and a Thelohanellus species infecting Carassius auratus gibelio (Bloch, 1782) were redescribed by their morphological, histological and molecular characterization. In the present study, the Myxobolus species infecting the muscle was identified as Myxobolus kingchowensis Chen et Ma, 1998 by the morphological and molecular data. Histologically, mature spores of M . kingchowensis were observed in the intercellular and connective tissue of muscle, though the plasmodia were not found. In addition, scattered spores also occurred in the intercellular of haematopoietic cells, intraepithelial of the renal tubules and interior of the melano-macrophage centres. Phylogenetic analysis showed that M . kingchowensis clustered in the clade of muscle-infecting Myxobolus species, further supporting muscle as the infection site of M . kingchowensis . The present Thelohanellus species infecting the gills was identified conspecific as Thelohanellus sinensis reported in Sun (2006) (mark it as T . sinensis -Sun)based on spore morphology, biological traits (host specificity and organ specificity), and molecular data. However, compared with the original description of T . sinensis Chen et Hsieh, 1960, the present Thelohanellus species and T . sinensis -Sun both infecting the gills of gibel carp are distinguishable from the original description in the host and infection site, which made the validity of T . sinensis -Sun dubious. Due to the absence of molecular data in the original description of T. sinensis , we suggest marking the present species and T . sinensis -Sun as T . cf. sinensis to avoid the confusion until T . sinensis is obtained from the type host and type infection site.

April 13, 2018

Cystic echinococcosis in animals and humans of Maharashtra State, India

V.M. Vaidya, R.J. Zende, A.M. Paturkar, M.L. Gatne, D.G. Dighe, R.N. Waghmare, S.L. Moon, S.S. Bhave, K.G. Bengale, N.V. Nikale

Page range: 232-243

Abstract

Echinococcosis/hydatidosis which is a neglected parasitic zoonosis in the developing country like India. The study was conducted during April, 2010 to March, 2017 to determine the prevalence among slaughtered food animals, dogs and human. The samples were collected from the various slaughterhouses situated in different regions of Maharashtra state. A total of 9464 cattle (male), 3661 buffalo, 47189 sheep, 33350 goats and 13579 pigs were scientifically examined during PM inspection at different slaughterhouses. The study revealed that the prevalence of disease in cattle (3.00%) was highest followed by buffalo (2.05%), pig (1.28%), sheep (0.09%) and goat (0.01%), by PM examination. The average estimated economic losses (direct and indirect) due to hydatidosis were Rs. 8,65,83,566 in cattle, Rs. 44,33,93,900 in buffalo, Rs. 7,24,50,615 in sheep, Rs. 1,88,29,359 in goat and Rs. 5,20,49,081 in pigs. Dog faecal samples analyzed and showed the prevalence of echinococcosis as 4.34% (19/438) by sedimentation method and positive samples were confirmed by PCR assay, whereas in high risk human, 11.09% sera samples were found to be positive for echinococcosis. However, based on data collection for seven years, 58 patients were found to be surgically operated for hydatid cyst removal. The results of the present study indicated that cystic echinococcosis/hydatidosis is prevalent in both human and animal population in study areas which attracts serious attention from veterinary and public health authority to reduce economic burden and in designing appropriate strategy for prevention and control of disease.

April 13, 2018

Structural predication and antigenic analysis of Toxoplasma gondii ROP20

Jian Zhou, Wanchun Wang, Pengxia Song, Lin Wang, Yali Han, Jingjing Guo, Zhen Hao, Xi Zhu, Qiong Zhou, Xiadong Du, Gang Lu, Shenyi He, Yingquan Luo

Page range: 244-251

Abstract

Toxoplasma gondii infects almost all the warm-blooded animals. ROP20 protein is expressed in the rhoptry of Toxoplasma gondii . In this study, the secondary structure of ROP20 was analyzed using SMART software. We constructed and analyzed the 3D model of ROP20 protein using SWISS-MODEL online procedure and Visual Molecular Dynamics (VMD) software. The structure analysis fully indicated that ROP20 protein is an important member of the ROP family. Furthermore, We used DNASTAR software and Epitope Database online service to analyze liner-B cell epitopes and T-cell epitopes of ROP20 protein. All the analysis results of ROP20 protein can provide positive information on treatment and vaccine for toxoplasmosis. Moreover, ROP20 gene was obtained from PCR, and a recombinant eukaryotic expression vector (pEGFP-C1-ROP20) was constructed in the following study. After restriction enzyme digestion, the constructed plasmid was transfected into HEK 293-T cells. The RT-PCR result indicated that the recombinant plasmid could transcribe successfully in HEK 293-T cell. The results of western blotting indicated the expressed proteins can be recognized by anti-STAg mouse sera.

April 13, 2018

Molecular cloning of enolase from Trichinella spiralis and the protective immunity in mice

Xuliang Zhang, Lixin Xu, Xiaokai Song, Xiangrui Li, Ruofeng Yan

Page range: 252-260

Abstract

Trichinella spiralis , the main pathogen of trichinosis, infects a wide range of mammalian hosts and is one of the most widespread parasites worldwide. For parasites, glycolysis is the most important way to generate energy. Previous studies showed that some enzymes involved in the glycolytic pathway play roles in regulation the host immunity. In this paper, enolase from T. spiralis was cloned and the protective potentials were studied. One hundred and sixty ICR mice were divided into four groups and vaccinated with recombinant enolase (pET-ENO), eukaryotic recombinant plasmid encoding enolase (pVAX1-ENO) and negative controls (pVAXl and PBS), respectively. Two weeks after the second immunization, each mouse was challenged orally with 200 muscle larvae (MLs) of T. spiralis . Results showed that mice vaccinated with pET-ENO and pVAX1-ENO induced specific antibodies of IgG, IgA, IgM, but no IgE. Subclasses of IgG antibodies showed that mice immunized with recombinant protein and recombinant plasmids induced a Th1/Th2 immune response. Concentrations of serum cytokines were detected and showed significant increase of IFN-γ, IL-4 and TGFβ1, while IL-17 in each group was not significantly different. Flow cytometric analysis showed significant increase of CD4+ and CD8+ T lymphocytes in the groups immunized with recombinant protein and recombinant plasmids. Challenge infection demonstrated that immunized groups had a reduced number of worm burdens. The reductions of larvae per gram muscle (LPG) in pET-ENO and pVAX1-ENO group were 17.7% and 15.8% when compared with PBS control.

April 13, 2018

A new species of Cacatuocotyle (Monogenea, Dactylogyridae) parasitizing Astyanax spp. (Characiformes, Characidae) from Brazil, including molecular data and a key to species identification

Aline Cristina Zago, Lidiane Franceschini, Maria Isabel Müller, Reinaldo José da Silva

Page range: 261-269

Abstract

The present study describes Cacatuocotyle papilionis n. sp. (Monogenea, Dactylogyridae) from the skin of the characid fishes Astyanax lacustris (Lütken, 1875) (= Astyanax altiparanae Garutti & Britski, 2000) and Astyanax fasciatus (Cuvier, 1819) (Characiformes, Characidae) from the Southeast of Brazil, supported by morphological and molecular data. The new species differs from all congeners, mainly due to the morphology of the ventral bar (resembling a butterfly), accessory piece, and the number of rings of the male copulatory organ (MCO), comprising a coiled tube with 4.5-5.5 counterclockwise rings. The first molecular data for this monogenean genus is provided in this study, using the partial sequences of the ribosomal gene (28S), as well as providing an identification key to the species.

April 13, 2018

Entamoeba histolytica L220 induces the in vitro activation of macrophages and neutrophils and is modulated by neurotransmitters

Fabiola del Rocío Villalobos-Gómez, Mario García-Lorenzana, Galileo Escobedo, Patricia Talamás-Rohana, Rogelio Salinas-Gutiérrez, Verónica-Ivonne Hernández-Ramírez, Esperanza Sánchez-Alemán, María del Rosario Campos-Esparza, Martín Humberto Muñoz-Ortega, Javier Ventura-Juárez

Page range: 270-279

Abstract

The neuroimmunoregulation of inflammation has been well characterized. Entamoeba histolytica provokes an inflammatory response in the host in which macrophages and neutrophils are the first line of defense. The aim of this study was to analyze the effect of the 220 kDa lectin of Entamoeba histolytica on stimulation of human macrophages and neutrophils, especially the secretion of cytokines and the relation of these to neurotransmitters. Human cells were interacted with L220, epinephrine, nicotine, esmolol and vecuronium bromide. The concentrations of IL-1β, IFN-γ, TNF-α and IL-10 were determined by ELISA at, 4 h of interaction. L220 has a cytokine stimulating function of macrophages and neutrophils for secretion of IL-1β, and IL-10 only by macrophages, which was modulated by the effect of vecuronium on cholinergic receptors in this immune cells.

April 13, 2018

Characterization of the complete mitochondrial genome of Metastrongylus salmi (M. salmi) derived from Tibetan pigs in Tibet, China

Kun Li, Muhammad Shahzad, Hui Zhang, Khalid Mehmood, Xiong Jiang, Houqiang Luo, Lihong Zhang, Xiaoqian Dong, Jiakui Li

Page range: 280-286

Abstract

The present study was designed to determine and analyze the mt genomes of Metastrongylus salmi ( M . salmi ), and reveal the phylogenetic relationships of this parasite using mt DNA sequences. Results showed that the complete mt genome of M . salmi was 13722 bp containing 12 protein-coding genes (cox1-3, nad1-6, nad4L, atp6 and cytb), 22 transfer RNA genes, and 2 ribosomal RNA genes (rrnL and rrnS). The overall A+T content was 73.54% and the nucleotide composition was A (23.52%), C (6.14%), G (19.60%), T (50.02%), and N (UCAG) (0.73%). A total of 4237 amino acids are encoded from the Tibetan isolates of M. salmi mt genomes. The ATA was predicted as the most common starting codon with 41.7% (5/12 protein genes); and 11 of the 12 protein genes were found to have a TAG or TAA translation termination codon. By clustering together the phylogenetic trees of Tibetan M . salmi and Austrian M . salmi , the M . salmi isolated from Tibetan pigs was found to be highly homological with that stemmed from Austrian one. This information provides meaningful insights into the phylogenetic position of the M . salmi China isolate and represents a useful resource for selecting molecular markers for diagnosis and population studies.

April 13, 2018

Balantioides coli: morphological and ultrastructural characteristics of pig and non-human primate isolates

Alynne da Silva Barbosa, Helene Santos Barbosa, Sandra Maria de Oliveira Souza, Laís Verdan Dib, Claudia Maria Antunes Uchôa, Otilio Machado Pereira Bastos, Maria Regina Reis Amendoeira

Page range: 287-298

Abstract

Balantioides coli is a ciliated protozoon that inhabits the intestine of pigs, non-human primates and humans. Light microscopy studies have described over 50 species of the genus Balantioides but their validity is in doubt. Due to the limited information about this genus, this study is aimed to identify morphological characteristics of Balantioides coli isolated using fluorescence microscopy and both scanning (SEM) and transmission electron microscopy (TEM). Trophozoites isolated from the feces of pig and macaque were washed and subjected to centrifugation. These cells were fixed with paraformaldehyde for immunofluorescence. Other aliquots of these trophozoites were fixed with glutaraldehyde, post fixed with osmium tetroxide and processed for SEM and TEM. Immunofluorescence studies revealed microtubules with a longitudinal distribution to the main axis of the parasite and in the constitution of cilia. SEM demonstrated a high concentration of cilia covering the oral apparatus and a poor presence of such structures in cytopyge. TEM revealed in the plasma membrane, several associated structures were observed to delineate the cellular cortex and mucocysts. The cytoskeleton of the oral region was observed in detail and had an organization pattern consisting of microtubules, which formed files and nematodesmal networks. Organelles such as hydrogenosomes like and peroxisomes were observed close to the cortex. Macronuclei were observed, but structures that were consistent with micronuclei were not identified. Ultrastructural morphological analysis of isolates confirms its similarity to Balantioides coli . In this study were identified structures that had not yet been described, such as hydrogenosomes like and cytoskeletal structures.

April 13, 2018

Genetic diversity of Taenia hydatigena in the northern part of the West Bank, Palestine as determined by mitochondrial DNA sequences

Kamel Adwan, Alaa Jayousi, Sameh Abuseir, Ibrahim Abbasi, Ghaleb Adwan, Naser Jarrar

Page range: 299-303

Abstract

Cysticercus tenuicollis is the metacestode of canine tapeworm Taenia hydatigena , which has been reported in domestic and wild ruminants and is causing veterinary and economic losses in the meat industry. This study was conducted to determine the sequence variation in the mitochondrial cytochrome c oxidase subunit 1 ( coxl ) gene in 20 isolates of T . hydatigena metacestodes (cysticercus tenuicollis) collected from northern West Bank in Palestine. Nine haplotypes were detected, with one prevailing (55%). The total haplotype diversity (0.705) and the total nucleotide diversity (0.0045) displayed low genetic diversity among our isolates. Haplotype analysis showed a star-shaped network with a centrally positioned common haplotype. The Tajima’s D, and Fu and Li’s statistics in cysticercus tenuicollis population of this region showed a negative value, indicating deviations from neutrality and both suggested recent population expansion for the population. The findings of this study would greatly help to implement control and preventive measures for T . hydatigena larvae infection in Palestine.

April 13, 2018

Patterns of the parasite communities in a fish assemblage of a river in the Brazilian Amazon region

Raimundo Rosemiro Jesus Baia, Alexandro Cezar Florentino, Luís Maurício Abdon Silva, Marcos Tavares-Dias

Page range: 304-316

Abstract

This paper characterizes the pattern of ectoparasite and endoparasite communities in an assemblage of 35 sympatric fish from different trophic levels in a tributary from the Amazon River system, northern Brazil. In detritivorous, carnivorous, omnivorous and piscivorous hosts, the species richness consisted of 82 ectoparasites and endoparasites, but protozoan ectoparasites such as Ichthyophthirius multifiliis , Piscinoodinium pillulare and Tripartiella sp. were dominant species predominated, such that they were present in 80% of the hosts. The taxon richness was in the following order: Monogenea > Nematoda > Digenea > Crustacea > Protozoa > Acanthocephala = Cestoda > Hirudinea. Among the hosts, the highest number of parasitic associations occurred in Satanoperca jurupari , Aequidens tetramerus , Hoplerythrinus unitaeniatus , Hoplosternum littorale , Cichlasoma amazonarum , Chaetobranchus flavescens , Squaliforma emarginata , Chaetobranchopsis orbicularis and Hoplias malabaricus . A weak positive correlation between ectoparasite abundance and length of the hosts was observed. Ectoparasite communities of detritivorous, carnivorous and omnivorous hosts were similar, but these differed from the communities of piscivorous hosts. Larval endoparasite species with low host specificity were the main determinants of the parasite infracommunity structure of the fish assemblage. Fish assemblage had few species of helminth that were specialist endoparasites, while many were parasites at the larval stage, infecting intermediate and paratenic hosts. Finally, carnivorous and omnivorous hosts harbored endoparasite communities that were more heterogeneous than those of detritivorous and piscivorous hosts. This result lends supports to the notion that the feeding habits of the host species are a significant factor in determining the endoparasites fauna.

April 13, 2018

Histological description of Cercaria etgesiiShchenkov, 2017 daughter sporocysts (Trematoda: Pleurogenidae): compartmentalization of the brood cavity and germinal mass morphology

Georgii A. Kremnev, Sergei V. Shchenkov

Page range: 317-324

Abstract

Morphology of the daughter sporocysts of Cercaria etgesii Shchenkov, 2017 (Pleurogenidae) has been described according to conventional histological techniques and subsequent 3D reconstruction with Bitplanelmaris® software. Novel data were obtained concerning the spatial organization of the brood cavity of Pleurogenidae daughter sporocysts. It was shown that the laminated structures of the endocyst are lining the brood cavity and carry out its compartmentalisation by formation of the chambers around developing and disintegrating embryos of cercariae. First description of the germinal mass of the Pleurogenidae daughter sporocysts is given. No mitoses in germinal mass were evident although apparent production of the cercariae embryos by the sporocysts take place. The analogous principle of daughter sporocysts morphofunctional organisation was noted for the studied representatives of closely related taxa of trematodes: Pleurogenidae ( C . etgesii ) and Microphallidae ( Microphallus ‘pygmaeus’ group).

April 13, 2018

Isolation, identification and characterization of the nematophagous fungus Arthrobotrys (Monacrosporium) sinense from China

Ya-Juan Xue, Er-Le Li, Cai-Xia Jing, Li Ma, Kui-Zheng Cai

Page range: 325-332

Abstract

With the development of anthelmintic resistance of parastic nematodes, it is necessary to isolate and study nematophagous fungi to screen out the native isolates for their potential in the biocontrol of domestic animal nematodosis. This study aimed to isolate the Arthrobotrys sinense ( Monacrosporium sinense ) of nematophagous fungus, to characterize representative molecular isolates using scanning electron microscope (SEM), and to determine the effect of the temperature and pH values on radial growth of the isolate. Five isolates were isolated from 1532 samples of different types, and their occurrence frequencies were 0.32% of the total samples. They were identified as A. sinense by means of morphology and the sequence of the 5.8S, 18S, and 28S rDNA, as well as internal transcribed spacers 1 and 2. The isolate NBS003 could grow from 11°C to 35°C and had optimal growth at 30°C. The isolate could grow at pH 4 to 11, and its optimal value was obtained at pH 9. SEM results showed that 6 h after their addition, the second stage larvae (L2) and the third stage infective larvae (L3) of Haemonchus contortus were captured. L2 and L3 were penetrated by the fungus at 18 and 24 h post-capture, respectively. L2 and L3 were completely digested at 84 and 90 h post-capture, respectively. The NBS003 of the A. sinense should have a certain potential to be used for capturing the free-living stage of nematodes in sheep.

April 13, 2018

First molecular characterization and morphological aspects of Sarcocystis fusiformis infecting water buffalo Bubalus bubalis in Egypt

Kareem Morsy, Fathy Abdel-Ghaffar, Saad Bin Dajem, Rewaida Abdel-Gaber, Fatma El Gazar

Page range: 333-345

Abstract

Fresh muscle samples from water buffalo ( Bubalus bubalis ) aged 2–15, from Giza Province, Egypt; were examined for Sarcocystis infection. Macroscopic ovoid sarcocysts embedded in the muscle tissues of the examined buffaloes were detected; they measured 152–230 (210 ± 7) μm in length and 37–119 (95 ± 3) μm in width. The esophagus was the most infected organ followed by the diaphragm, and tongue, while the heart muscles were the least infected. The cyst cavity was compartmentalized by septa derived from the ground substance located under the primary cyst wall. Using transmission electron microscopy, the primary cyst wall bordered sarcocysts were determined to be 0.08–0.22 μm in thickness, raised from the parasitophorous vacuolar membrane, and surrounded by a secondary cyst wall of host origin. The primary cyst wall had irregular wall folds with numerous cauliflower-like projections of variable sizes and shapes accompanied by knob-like electron-dense elevations. 18S rRNA gene expression studies confirmed that the present parasite isolates belonged to the genus Sarcocystis . The sequence data showed significant identities (>90%) with archived gene sequences from many Eimeriidae organisms, and a dendogram showing the phylogenetic relationship was constructed. The most closely related species was Sarcocystis fusiformis KR186117, with an identity percentage of 98%. The recovered sequences were deposited in the GenBank under the accession number MG572125. The present study, to our knowledge, is the first collective ultrastructural and molecular study that confirmed the taxonomy of sarcocysts isolated from water buffaloes in Egypt as Sarcocystis fusiformis .

April 13, 2018

Prevalence and risk factors associated with Tritrichomonas foetus infection in cattle in the state of Paraíba, Brazil

Ruy Brayner de Oliveira Filho, Karla Campos Malta, Jonas de Melo Borges, Pollyanne Raysa Fernandes de Oliveira, Givanildo Jacinto dos Santos Filho, Glaucia Grazielle Nascimento, Rinaldo Aparecido Mota, José Wilton Pinheiro Júnior

Page range: 346-353

Abstract

The objective of this study was to determine the prevalence of Tritrichomonas foetus infection and to evaluate risk factors associated with this infection among cattle in the state of Paraíba in northeastern Brazil. Samples of cervicovaginal mucus from 290 females and smegma from 59 males [beef, 31; mixed aptitude (beef and dairy), 10; and dairy, 18] from 31 farms were collected. Modified Diamond’s medium and polymerase chain reaction (PCR) were used for the laboratory diagnosis of T. foetus infection. Univariate analysis and logistic regression were performed to test for potential risk factors in addition to prevalence mapping. No sample was positive for T. foetus in culture, and the prevalence of T. foetus infection using PCR was 3.7% (13/349) [confidence interval (CI) 95%, 2.1%–6.4%]. In total, 19.3% (6/31) of the farms had at least one animal positive for T. foetus . The contact of females with males from other farms [Odds ratio, 5.9; 95% CI, 1.5–22.4; p = 0.009] was identified as a risk factor for T. foetus infection. This study demonstrates that T. foetus infection is prevalent among dairy cows in the state of Paraíba, Brazil. Sexual resting, removal of positive females, and avoiding contact of females with males from other farms are recommended to reduce the risk of infection.

April 13, 2018

The development of Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae) in its intermediate host Cypridopsis vidua in Brazil

Felipe de Sousa Lourenço, Germán Augusto Murrieta Morey, José Celso de Oliveira Malta

Page range: 354-359

Abstract

The family Neoechinorhynchidae includes seven species of Neoechinorhynchus from freshwater fishes of Brazil. Although several Neoechinorhynchus species are cited infecting different fish species in Brazil, there is a lack of information concerning to their life cycle and the identification of the intermediate hosts. Thus, the aim of the present study was to describe the development of Neoechinorhynchus buttnerae in its intermediate host collected in a fish farm located in Rio Preto da Eva, Amazonas, Brazil. To verify the presence of N. buttnerae in the fish pond, twenty Colossoma macropomum were captured and analyzed, being corroborated the presence of this parasite species. Samples of plankton were also collected, finding the ostracod Cypridopsis vidua as the intermediate host. For the description of the larvae development, a laboratory experimental procedure was conducted by feeding the collected ostracods with the eggs of the adult specimens taken from the sampled fish. To observe the stages of development an artificial hatch was performed. Every stage of development was photographed, measured, drawn and described. The time of development of the immature stages of N. buttnerae was 29 days, reporting the stages: acanthor, acanthella (with eight developmental changes) and cystacanth. As high infections by N. buttnerae causes morphological damages to the intestine and may compromise the quality of C. macropomum and in consequence the production of fish farmers in the Brazilian Amazon region, the knowledge of its intermediate host and the understanding of its life cycle represents a useful information to prevent and combat infections by this parasite.

April 13, 2018

Primary culture of cat intestinal epithelial cells in vitro and the cDNA library construction

Gui hua Zhao, Ye Liu, Yun tang Cheng, Qing song Zhao, Xiao Qiu, Chao Xu, Ting Xiao, Song Zhu, Gong zhen Liu, Kun Yin

Page range: 360-367

Abstract

Felids are the only definitive hosts of Toxoplasma gondii . To lay a foundation for screening the T . gondii -felids interaction factors, we have developed a reproducible primary culture method for cat intestinal epithelial cells (IECs). The primary IECs were isolated from a new born cat’s small intestine jejunum region without food ingress, and respectively in vitro cultured by tissue cultivation and combined digestion method with collagenase XI and dispase I, then purified by trypsinization. After identification, the ds cDNA of cat IECs was synthesized for constructing pGADT7 homogenization three-frame plasmid, and transformed into the yeast Y187 for generating the cDNA library. Our results indicated that cultivation of primary cat IECs relays on combined digestion to form polarized and confluent monolayers within 3 days with typical features of normal epithelial cells. The purified cells cultured by digestion method were identified to be nature intestinal epithelial cells using immunohistochemical analysis and were able to maintain viability for at least 15 passages. The homogenizable ds cDNA, which is synthesized from the total RNA extracted from our cultured IECs, distributed among 0.5–2.0 kb, and generated satisfying three-frame cDNA library with the capacity of 1.2 × 10 6 and the titer of 5.2 × 10 7 pfu/mL. Our results established an optimal method for the culturing and passage of cat IECs model in vitro , and laid a cDNA library foundation for the subsequent interaction factors screening by yeast two-hybrid.

April 13, 2018

Evaluation of the protective effect of a prime-boost strategy with plasmid DNA followed by recombinant adenovirus expressing BmAMA1 as vaccines against Babesia microti infection in hamster

Guanbo Wang, Longzheng Yu, Artemis Efstratiou, Paul Franck Adjou Moumouni, Mingming Liu, Huanping Guo, Yang Gao, Shinuo Cao, Mo Zhou, Jixu Li, Aaron Edmond Ringo, Xuenan Xuan

Page range: 368-374

Abstract

In the present study, we have investigated the protective effect of a heterologous prime-boost strategy with priming plasmid DNA followed by recombinant adenovirus, both expressing BmAMA1, against Babesia microti infection. Four groups consisting of 3 hamsters per group were immunized with pBmAMA1/Ad5BmAMA1, pNull/Ad5BmAMA1, pBmAMA1/Ad5Null and pNull/Ad5Null, followed by challenge infection with B . microti . Our results showed that hamsters immunized with plasmid and adenovirus expressing BmAMA1 developed a robust IgG and IgG2a antibody response against BmAMA1, suggesting the DNA vaccine or viral vector vaccine tend to induce a Th1-biased response. Compared to the control hamsters, the hamsters vaccinated either with the prime-boost strategy or one of the two “vaccines” exhibited no significant protection against B . microti challenge. Although a slight difference in terms of parasitemia and hematocrit values at days 14–16 post challenge infection was observed, no other statistical difference was detected. Our results indicate that the prime-boost vaccination strategy of injection of plasmid and adenovirus expressing BmAMA1 is not efficient in protecting against B . microti infection.

April 13, 2018

A new species of Chondracanthus (Cyclopoida: Chondracanthidae) parasitic on deep-sea Dibranchus spongiosa(Lophiiformes: Ogcocephalidae) from the Eastern Central Pacific

Samuel Gómez, Hugo Aguirre-Villaseñor, Francisco Neptali Morales-Serna

Page range: 375-385

Abstract

A total of 228 sampling stations were visited for benthic fauna during a series of oceanographic cruises in the Gulf of California, west coast of the Baja California Peninsula, and Eastern Central Pacific from year 1991 to 2014. Among others, three fish species of the genus Dibranchus were caught in 28 stations. Of these, D . spongiosa was the most common and abundant. Close inspection of this fish revealed the presence of a new species of parasitic copepod, Chondracanthus dibranchi sp. nov., found in the gill cavity of seven specimens of D . spongiosa . Chondracanthus dibranchi sp. nov. seems to be morphologically related to C . psetti and C . janebennettae . The female of C . janebennettae can be separated from these other two congeners by the general shape of the head and abdomen, by the number of teeth on the mandibular blade, and by the general body shape. Chondracanthus psetti and C . dibranchi sp. nov. share the relative lengths of legs 1 and 2, the relative size and shape of the genito-abdomen, and the conical attenuating lateral processes on the trunk of the female. The females of these two species can be separated by the shape and armature of the antennule, shape of the antenna, the claw of the maxilliped, the rami of leg 1 and 2 and posterior processes, the head region, and by the position of the lateral processes of the trunk. An amendment to Tang’s (2007) key to the species of Chondracanthus is proposed.

April 13, 2018

Evaluation of four commercial DNA extraction kits for the detection of Microsporidia and the importance of pretreatments in DNA isolation

Ülfet Çetinkaya, Arzuv Charyyeva, Eda Sivcan, Esra Gürbüz

Page range: 386-392

Abstract

Microsporidia are obligate intracellular parasitic protozoa infecting the wide variety of hosts and are commonly known as a cause of chronic diarrhea particularly in immunocompromised individuals. Molecular-based tests have high sensitivity and specificity in disease diagnosis. However, these tests’ performance relies on the isolation of DNA in a good concentration. The standard procedures of commercial DNA extraction kits are usually insufficient for this purpose due to the tough walls of spores. This study aimed to test the significance of pretreatments by glass beads and freeze-thawing processes in DNA isolation from microsporidia spores. The parasite was cultured in growing Vero cells and seven serial dilutions were prepared from the collected spores. DNA purification was performed according to different tissue kits and stool kit procedures with and without any pretreatment. Concentration of isolated DNA samples were evaluated by real-time PCR. As a result of this study, the detectable amount of spores is minimum 10 spores in each 100 μ! sample according to the different tissue kits’ standard protocols. However, according to the DNA stool mini kit, the detectable amount of spores was found to be 1,000 spores/100 μl of stool sample when pretreated with both the freeze-thawing and glass beads methods.In conclusion, the current study demonstrated that further pretreatments are an essential process for DNA extraction from the stool specimens in order to avoid possible false negativity in the diagnosis of microsporidiosis.

April 13, 2018

Presence of Acanthamoeba in the ocular surface in a Spanish population of contact lens wearers

Javier Rodríguez-Martín, Pedro Rocha-Cabrera, María Reyes-Batlle, Atteneri López-Arencibia, Ines Sifaoui, Aitor Rizo-Liendo, Carlos J. Bethencourt-Estrella, José E. Piñero, Jacob Lorenzo-Morales

Page range: 393-396

Abstract

Pathogenic strains of Acanthamoeba are causative agents of a sight-threatening infection of the cornea known as Acanthamoeba keratitis (AK) which mainly affects contact lens wearers and it is commonly related to poor hygiene of contact lenses and their cases. Moreover, treatment of AK is complex due to the existence of a highly resistant cyst stage and if not diagnosed early has poor prognosis, leading to blindness and/or keratoplasty. Even though AK is increasing worldwide as well as awareness among patients and clinicians, it is still a poorly studied pathogen. Additionally, a remaining question to be answered is whether these opportunistic pathogens are present in the ocular surface of healthy contact lens wearers since they are the main group at risk.In order to carry out this study, sterile Schirmer strip tests were collected from a group of individuals all of them contact lens wearers who were attending a local ophthalmology clinic in Tenerife, Canary Islands, Spain. The collected samples (100 eyes of 50 patients) were cultured in 2% Non-Nutrient Agar (NNA) plates and positive plates (16) were then cultured in axenic conditions for further analyses. Molecular analysis classified all isolated strains belonged to Acanthamoeba genotype T4 and osmotolerance and thermotolerance assays revealed that all strains were potentially pathogenic. In conclusion, the ocular surface of contact lens wearers included in this study was colonized by potentially pathogenic strains of Acanthamoeba and should be considered as a risk for AK infection in this region and worldwide.

April 13, 2018

Genotyping and virulence analysis of Toxoplasma gondii isolates from a dead human fetus and dead pigs in Jiangsu province, Eastern China

Zhaofeng Hou, Yonghua Zhou, Dandan Liu, Shijie Su, Zhenxing Zhao, Jinjun Xu, Jianping Tao

Page range: 397-411

Abstract

Toxoplasma gondii is an obligate intracellular parasite with worldwide distribution. Virulence of T . gondii is a multigenic trait. Genetic and virulence data for T . gondii isolates from humans and animals in China have been reported. However, almost all biological materials used for genotyping of T . gondii from humans and pigs were DNA samples prepared from tissues, and T. gondii strains used for virulence analysis were isolated mainly from cats. In this study, one isolate from a dead human fetus was identified as type I (ToxoDB #10) while the two isolates from dead pigs were type Chinese I (ToxoDB #9) with PCR-restriction fragment length polymorphism using 10 markers ( SAG1 , SAG2 , SAG3 , BTUB , GRA6 , c22 – 8 , c29 – 2 , L358 , PK1 and Apico). Three isolates were comfirmed as virulent strains in mice. By cloning and sequences analysis, all isolates contained a Pvu II restriction site (572–577 bp) in the KHB fragment and five tandem repeats in the 5′ UTR region of SAG1 , which were associated with T . gondii virulence. The type Chinese I isolates contained two deletions of 15 and 3 bp at positions 635 to 649 and 658 to 660 in the GRA6 , which were correlated with genotype, but not with virulence. To our knowledge, this is the first report on the systematic analysis of murine virulence of type Chinese I strain from pigs, and the associations of sequences of the KHB fragment and SAG1 with virulence of type Chinese I strain. The Chinese I genotype was more closely related to type II strains.

Research note

April 13, 2018

Detection of Dirofilaria immitis antigen and antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi and Ehrlichia canis in dogs from ten provinces of China

Jiawei Wang, Patrick Kelly, Jilei Zhang, Zhengsheng Shi, Chunlian Song, Xiaofeng Zheng, Yi Zhang, Yongqing Hao, Haiju Dong, Heba S. El-Mahallawy, Wenbin Xiong, Heng Wang, Jianji Li, Xinjun Zhang, Chengming Wang

Page range: 412-415

Abstract

Despite the fact vector-borne diseases (VBDs) have been increasingly reported in dogs worldwide, there are only limited reports on VBDs in dogs in China with most being based on molecular detection of active infections. To provide further data on the exposure of dogs in China to VBD agents, we used commercial immunochromatographic assays to test plasma from 637 apparently healthy indoor and breeding colony dogs from 21 veterinary clinics in 10 provinces in China and a commercial dog breeding facility for circulating antigen of Dirofilaria immitis , and for circulating antibodies against Ehrlichia spp., Anaplasma spp., and Borrelia burgdorferi . Overall, we found only low levels of exposure to Ehrlichia spp. (4.7%; 30/637), Anaplasma spp. (1.4%; 9/637), B . burgdorferi (0.9%; 6/637) and D . immitis (0.2%; 1/637) with most of the positive animals coming from the commercial breeding colony (26/103; 25.2%) where ectoparasites were most commonly noted. At least one vector-borne agent was found in dogs from 6 of the 10 provinces investigated. Our results confirm exposure of dogs from around China to a variety of VBDs, even indoor pets seldom observed to harbor ectoparasites.

Research note

April 13, 2018

Evaluation of frequency of antibodies against Toxoplasma gondii,Neospora caninum and Sarcocystis spp. and transmission routes in sheep from Humid Pampa, Argentina

Yanina P. Hecker, Fernando Mogaburu Masson, Joaquín I. Armendano, Juan Cora, Carlos Flores Olivares, Ignacio Gual, Lais Pardini, Dadín P. Moore, Gastón Moré, Germán J. Cantón

Page range: 416-421

Abstract

The aim of this study was to describe the frequency of ovine specific antibodies to Toxoplasma gondii , Neospora caninum and Sarcocystis spp. and to estimate different transmission routes of these infections. One hundred and thirty Texel sheep and their 117 Texel lambs were included in the study. Serum samples were tested for antibodies to T . gondii , N . caninum and Sarcocystis spp. using IFAT. Toxoplasma gondii seroprevalence was 10.00% in sheep (IC 95% : 4.80–15.20%), being higher in adult sheep (≥12 year) than in younger sheep (OR 1.30; 95% CI, 1.10–1.50). N . caninum and Sarcocystis spp. seroprevalences were 1.54% (IC 95% : 0.00–5.70) and 72.09% (IC 95% : 67.70–82.70), respectively, with no association between age and seropositivity in sheep ( P >0.05). T . gondii seroprevalence in lambs was 4.27% (IC 95% : 0.61–7.94). No association between T . gondii serological status in sheep and their lambs was detected ( P = 0.07). Two T . gondii and Sarcocystis spp. seropositive lambs were euthanized and T . gondii and Sarcocystis spp. DNA was detected by PCR in their tissues. In conclusion, the increase of T . gondii seropositivity in relationship with sheep age and the lack of association between sheep-lamb serological status, suggest that horizontal infection is the main transmission route in this flock as reported before. Due to the low number of N . caninum -seropositive ewes no assumptions can be done about the impact of this parasite in this flock. According with previous reports, the main transmission route for Sarcocystis spp. in this species in the present study was horizontal.

Case report

April 13, 2018

First report of Cryptosporidium parvum in a dromedary camel calf from Western Australia

Alireza Zahedi, Gary K.C. Lee, Telleasha L. Greay, Audra L. Walsh, David J.C. Blignaut, Una M. Ryan

Page range: 422-427

Abstract

Cryptosporidium is an important enteric parasite that can contribute large numbers of infectious oocysts to drinking water catchments. As a result of its resistance to disinfectants including chlorine, it has been responsible for numerous waterborne outbreaks of gastroenteritis. Wildlife and livestock play an important role in the transmission of Cryptosporidium in the environment. Studies conducted outside Australia have indicated that camels may also play a role in the transmission of zoonotic species of Cryptosporidium . Despite Australia being home to the world’s largest camel herd, nothing is known about the prevalence and species of Cryptosporidium infecting camels in this country. In the present study, C. parvum was identified by PCR amplification and sequencing of a formalin-fixed intestinal tissue specimen from a one-week old dromedary camel ( Camelus dromedarius ). Subtyping analysis at the glycoprotein 60 ( gp60 ) locus identified C. parvum subtype IIaA17G2R1, which is a common zoonotic subtype reported in humans and animals worldwide. Histopathological findings also confirmed the presence of large numbers of variably-sized (1–3 µm in diameter) circular basophilic protozoa – consistent with Cryptosporidium spp.– adherent to the mucosal surface and occasionally free within the lumen. Further analysis of the prevalence and species of Cryptosporidium in camel populations across Australia are essential to better understand their potential for contamination of drinking water catchments.

Case report

April 13, 2018

Virulent T4 Acanthamoeba causing keratitis in a patient after swimming while wearing contact lenses in Southern Brazil

Laura Fuhrich Fabres, Vinicius José Maschio, Denise Leal dos Santos, Sergio Kwitko, Diane Ruschel Marinho, Bruno Schneider de Araújo, Claudete Inês Locatelli, Marilise Brittes Rott

Page range: 428-432

Abstract

Several strains of free-living amoebae belonging to the genus Acanthamoeba can cause a painful sight-threatening disease of the cornea known as Acanthamoeba keratitis (AK). The numbers of AK cases keep rising worldwide mainly due to an increase in contact lens wearers and lack of hygiene in the maintenance of contact lenses and their cases. We report a case of AK in a healthy young woman admitted to the Hospital de Clinicas in Porto Alegre, southern Brazil. Corneal scrapings were examined for the presence of Acanthamoeba strains. The initial isolate was characterized by morphological and genotypic properties. The isolate belonged to group III according to Pussard and Pons’ cyst morphology. Analysis of its 18S rDNA sequence identified the isolate as genotype T4. The T4 genotype is the most commonly reported among keratitis isolates and the most common in environmental samples.

Erratum

April 13, 2018

Transovarial persistence of Babesia ovata DNA in a hard tick, Haemaphysalis longicornis, in a semi-artificial mouse skin membrane feeding system

Rika Umemiya-Shirafuji, Takeshi Hatta, Kazuhiro Okubo, Moeko Sato, Hiroki Maeda, Aiko Kume, Naoaki Yokoyama, Ikuo Igarashi, Naotoshi Tsuji, Kozo Fujisaki, Noboru Inoue, Hiroshi Suzuki

Page range: 433-433

Erratum

April 13, 2018

Erratum

Page range: 434-434

About this journal

This journal is now published by Springer.