bluuebunny/crossref_metadata_2025 huggingface dataset:
Cystic echinococcosis in animals and humans of Maharashtra State, India
Authors: Vaidya V.M. , Zende R.J. , Paturkar A.M. , Gatne M.L. , Dighe D.G. , Waghmare R.N. , Moon S.L. , Bhave S.S. , Bengale K.G. , Nikale N.V.
Published: April 2018

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.
Structural predication and antigenic analysis of Toxoplasma gondii ROP20
Authors: Zhou Jian , Wang Wanchun , Song Pengxia , Wang Lin , Han Yali , Guo Jingjing , Hao Zhen , Zhu Xi , Zhou Qiong , Du Xiadong , Lu Gang , He Shenyi , Luo Yingquan
Published: April 2018

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.
Molecular cloning of enolase from Trichinella spiralis and the protective immunity in mice
Authors: Zhang Xuliang , Xu Lixin , Song Xiaokai , Li Xiangrui , Yan Ruofeng
Published: April 2018

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.
A new species of Cacatuocotyle (Monogenea, Dactylogyridae) parasitizing Astyanax spp. (Characiformes, Characidae) from Brazil, including molecular data and a key to species identification
Authors: Zago Aline Cristina , Franceschini Lidiane , Müller Maria Isabel , Silva Reinaldo José da
Published: April 2018

The present study describesCacatuocotyle papilionisn. sp. (Monogenea, Dactylogyridae) from the skin of the characid fishesAstyanax lacustris(Lütken, 1875) (=Astyanax altiparanaeGarutti & Britski, 2000) andAstyanax 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.
Entamoeba histolytica L220 induces the in vitro activation of macrophages and neutrophils and is modulated by neurotransmitters
Authors: Villalobos-Gómez Fabiola del Rocío , García-Lorenzana Mario , Escobedo Galileo , Talamás-Rohana Patricia , Salinas-Gutiérrez Rogelio , Hernández-Ramírez Verónica-Ivonne , Sánchez-Alemán Esperanza , Campos-Esparza María del Rosario , Muñoz-Ortega Martín Humberto , Ventura-Juárez Javier
Published: April 2018

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.
Characterization of the complete mitochondrial genome of Metastrongylus salmi (M. salmi) derived from Tibetan pigs in Tibet, China
Authors: Li Kun , Shahzad Muhammad , Zhang Hui , Mehmood Khalid , Jiang Xiong , Luo Houqiang , Zhang Lihong , Dong Xiaoqian , Li Jiakui
Published: April 2018

The present study was designed to determine and analyze themtgenomes ofMetastrongylus salmi(M.salmi), and reveal the phylogenetic relationships of this parasite usingmtDNA sequences. Results showed that the completemtgenome ofM.salmiwas 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 ofM. salmi mtgenomes. 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 TibetanM.salmiand AustrianM.salmi, theM.salmiisolated 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 theM.salmiChina isolate and represents a useful resource for selecting molecular markers for diagnosis and population studies.
Balantioides coli: morphological and ultrastructural characteristics of pig and non-human primate isolates
Authors: Barbosa Alynne da Silva , Barbosa Helene Santos , Souza Sandra Maria de Oliveira , Dib Laís Verdan , Uchôa Claudia Maria Antunes , Bastos Otilio Machado Pereira , Amendoeira Maria Regina Reis
Published: April 2018

Balantioides coliis a ciliated protozoon that inhabits the intestine of pigs, non-human primates and humans. Light microscopy studies have described over 50 species of the genusBalantioidesbut their validity is in doubt. Due to the limited information about this genus, this study is aimed to identify morphological characteristics ofBalantioides coliisolated 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 toBalantioides coli. In this study were identified structures that had not yet been described, such as hydrogenosomes like and cytoskeletal structures.
Patterns of the parasite communities in a fish assemblage of a river in the Brazilian Amazon region
Authors: Baia Raimundo Rosemiro Jesus , Florentino Alexandro Cezar , Silva Luís Maurício Abdon , Tavares-Dias Marcos
Published: April 2018

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 asIchthyophthirius multifiliis,Piscinoodinium pillulareandTripartiellasp. 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 inSatanoperca jurupari,Aequidens tetramerus,Hoplerythrinus unitaeniatus,Hoplosternum littorale,Cichlasoma amazonarum,Chaetobranchus flavescens,Squaliforma emarginata,Chaetobranchopsis orbicularisandHoplias 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.
Histological description of Cercaria etgesii Shchenkov, 2017 daughter sporocysts (Trematoda: Pleurogenidae): compartmentalization of the brood cavity and germinal mass morphology
Authors: Kremnev Georgii A. , Shchenkov Sergei V.
Published: April 2018

Morphology of the daughter sporocysts ofCercaria etgesiiShchenkov, 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).
Isolation, identification and characterization of the nematophagous fungus Arthrobotrys (Monacrosporium) sinense from China
Authors: Xue Ya-Juan , Li Er-Le , Jing Cai-Xia , Ma Li , Cai Kui-Zheng
Published: April 2018

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 theArthrobotrys 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 asA. sinenseby 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) ofHaemonchus contortuswere 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 theA. sinenseshould have a certain potential to be used for capturing the free-living stage of nematodes in sheep.
First molecular characterization and morphological aspects of Sarcocystis fusiformis infecting water buffalo Bubalus bubalis in Egypt
Authors: Morsy Kareem , Abdel-Ghaffar Fathy , Bin Dajem Saad , Abdel-Gaber Rewaida , El Gazar Fatma
Published: April 2018

Fresh muscle samples from water buffalo (Bubalus bubalis) aged 2–15, from Giza Province, Egypt; were examined forSarcocystisinfection. 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 genusSarcocystis. 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 wasSarcocystis fusiformisKR186117, 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 asSarcocystis fusiformis.
Prevalence and risk factors associated with Tritrichomonas foetus infection in cattle in the state of Paraíba, Brazil
Authors: Filho Ruy Brayner de Oliveira , Malta Karla Campos , Borges Jonas de Melo , Oliveira Pollyanne Raysa Fernandes de , Filho Givanildo Jacinto dos Santos , Nascimento Glaucia Grazielle , Mota Rinaldo Aparecido , Júnior José Wilton Pinheiro
Published: April 2018

The objective of this study was to determine the prevalence ofTritrichomonas foetusinfection 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 ofT. foetusinfection. Univariate analysis and logistic regression were performed to test for potential risk factors in addition to prevalence mapping. No sample was positive forT. foetusin culture, and the prevalence ofT. foetusinfection 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 forT. 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 forT. foetusinfection. This study demonstrates thatT. foetusinfection 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.
The development of Neoechinorhynchus buttnerae (Eoacanthocephala: Neoechinorhynchidae) in its intermediate host Cypridopsis vidua in Brazil
Authors: Lourenço Felipe de Sousa , Morey Germán Augusto Murrieta , Malta José Celso de Oliveira
Published: April 2018

The family Neoechinorhynchidae includes seven species ofNeoechinorhynchusfrom freshwater fishes of Brazil. Although severalNeoechinorhynchusspecies 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 ofNeoechinorhynchus buttneraein its intermediate host collected in a fish farm located in Rio Preto da Eva, Amazonas, Brazil. To verify the presence ofN. buttneraein the fish pond, twentyColossoma macropomumwere captured and analyzed, being corroborated the presence of this parasite species. Samples of plankton were also collected, finding the ostracodCypridopsis viduaas 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 ofN. buttneraewas 29 days, reporting the stages: acanthor, acanthella (with eight developmental changes) and cystacanth. As high infections byN. buttneraecauses morphological damages to the intestine and may compromise the quality ofC. macropomumand 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.
Primary culture of cat intestinal epithelial cells in vitro and the cDNA library construction
Authors: Zhao Gui hua , Liu Ye , Cheng Yun tang , Zhao Qing song , Qiu Xiao , Xu Chao , Xiao Ting , Zhu Song , Liu Gong zhen , Yin Kun
Published: April 2018

Felids are the only definitive hosts ofToxoplasma gondii. To lay a foundation for screening theT.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 respectivelyin vitrocultured 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 × 106and the titer of 5.2 × 107pfu/mL. Our results established an optimal method for the culturing and passage of cat IECs modelin vitro, and laid a cDNA library foundation for the subsequent interaction factors screening by yeast two-hybrid.
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
Authors: Wang Guanbo , Yu Longzheng , Efstratiou Artemis , Moumouni Paul Franck Adjou , Liu Mingming , Guo Huanping , Gao Yang , Cao Shinuo , Zhou Mo , Li Jixu , Ringo Aaron Edmond , Xuan Xuenan
Published: April 2018

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, againstBabesia microtiinfection. Four groups consisting of 3 hamsters per group were immunized with pBmAMA1/Ad5BmAMA1, pNull/Ad5BmAMA1, pBmAMA1/Ad5Null and pNull/Ad5Null, followed by challenge infection withB.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 againstB.microtichallenge. 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 againstB.microtiinfection.
A new species of Chondracanthus (Cyclopoida: Chondracanthidae) parasitic on deep-sea Dibranchus spongiosa(Lophiiformes: Ogcocephalidae) from the Eastern Central Pacific
Authors: Gómez Samuel , Aguirre-Villaseñor Hugo , Morales-Serna Francisco Neptali
Published: April 2018

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 genusDibranchuswere caught in 28 stations. Of these,D.spongiosawas the most common and abundant. Close inspection of this fish revealed the presence of a new species of parasitic copepod,Chondracanthus dibranchisp. nov., found in the gill cavity of seven specimens ofD.spongiosa.Chondracanthus dibranchisp. nov. seems to be morphologically related toC.psettiandC.janebennettae. The female ofC.janebennettaecan 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 psettiandC.dibranchisp. 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 ofChondracanthusis proposed.
Evaluation of four commercial DNA extraction kits for the detection of Microsporidia and the importance of pretreatments in DNA isolation
Authors: Çetinkaya Ülfet , Charyyeva Arzuv , Sivcan Eda , Gürbüz Esra
Published: April 2018

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.
Presence of Acanthamoeba in the ocular surface in a Spanish population of contact lens wearers
Authors: Rodríguez-Martín Javier , Rocha-Cabrera Pedro , Reyes-Batlle María , López-Arencibia Atteneri , Sifaoui Ines , Rizo-Liendo Aitor , Bethencourt-Estrella Carlos J. , Piñero José E. , Lorenzo-Morales Jacob
Published: April 2018

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.
Genotyping and virulence analysis of Toxoplasma gondii isolates from a dead human fetus and dead pigs in Jiangsu province, Eastern China
Authors: Hou Zhaofeng , Zhou Yonghua , Liu Dandan , Su Shijie , Zhao Zhenxing , Xu Jinjun , Tao Jianping
Published: April 2018

Toxoplasma gondiiis an obligate intracellular parasite with worldwide distribution. Virulence ofT.gondiiis a multigenic trait. Genetic and virulence data forT.gondiiisolates from humans and animals in China have been reported. However, almost all biological materials used for genotyping ofT.gondiifrom humans and pigs were DNA samples prepared from tissues, andT. gondiistrains 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,PK1and Apico). Three isolates were comfirmed as virulent strains in mice. By cloning and sequences analysis, all isolates contained aPvuII restriction site (572–577 bp) in the KHB fragment and five tandem repeats in the 5′ UTR region ofSAG1, which were associated withT.gondiivirulence. The type Chinese I isolates contained two deletions of 15 and 3 bp at positions 635 to 649 and 658 to 660 in theGRA6, 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 andSAG1with virulence of type Chinese I strain. The Chinese I genotype was more closely related to type II strains.
Detection of Dirofilaria immitis antigen and antibodies against Anaplasma phagocytophilum, Borrelia burgdorferi and Ehrlichia canis in dogs from ten provinces of China
Authors: Wang Jiawei , Kelly Patrick , Zhang Jilei , Shi Zhengsheng , Song Chunlian , Zheng Xiaofeng , Zhang Yi , Hao Yongqing , Dong Haiju , El-Mahallawy Heba S. , Xiong Wenbin , Wang Heng , Li Jianji , Zhang Xinjun , Wang Chengming
Published: April 2018

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 ofDirofilaria immitis, and for circulating antibodies againstEhrlichiaspp.,Anaplasmaspp., andBorrelia burgdorferi. Overall, we found only low levels of exposure toEhrlichiaspp. (4.7%; 30/637),Anaplasmaspp. (1.4%; 9/637),B.burgdorferi(0.9%; 6/637) andD.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.
Evaluation of frequency of antibodies against Toxoplasma gondii,Neospora caninum and Sarcocystis spp. and transmission routes in sheep from Humid Pampa, Argentina
Authors: Hecker Yanina P. , Masson Fernando Mogaburu , Armendano Joaquín I. , Cora Juan , Olivares Carlos Flores , Gual Ignacio , Pardini Lais , Moore Dadín P. , Moré Gastón , Cantón Germán J.
Published: April 2018

The aim of this study was to describe the frequency of ovine specific antibodies toToxoplasma gondii,Neospora caninumandSarcocystisspp. 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 toT.gondii,N.caninumandSarcocystisspp. using IFAT.Toxoplasma gondiiseroprevalence was 10.00% in sheep (IC95%: 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.caninumandSarcocystisspp. seroprevalences were 1.54% (IC95%: 0.00–5.70) and 72.09% (IC95%: 67.70–82.70), respectively, with no association between age and seropositivity in sheep (P>0.05).T.gondiiseroprevalence in lambs was 4.27% (IC95%: 0.61–7.94). No association betweenT.gondiiserological status in sheep and their lambs was detected (P= 0.07). TwoT.gondiiandSarcocystisspp. seropositive lambs were euthanized andT.gondiiandSarcocystisspp. DNA was detected by PCR in their tissues. In conclusion, the increase ofT.gondiiseropositivity 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 ofN.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 forSarcocystisspp. in this species in the present study was horizontal.
First report of Cryptosporidium parvum in a dromedary camel calf from Western Australia
Authors: Zahedi Alireza , Lee Gary K.C. , Greay Telleasha L. , Walsh Audra L. , Blignaut David J.C. , Ryan Una M.
Published: April 2018

Cryptosporidiumis 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 ofCryptosporidiumin the environment. Studies conducted outside Australia have indicated that camels may also play a role in the transmission of zoonotic species ofCryptosporidium. Despite Australia being home to the world’s largest camel herd, nothing is known about the prevalence and species ofCryptosporidiuminfecting camels in this country. In the present study,C. parvumwas 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 identifiedC. parvumsubtype 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 withCryptosporidiumspp.– adherent to the mucosal surface and occasionally free within the lumen. Further analysis of the prevalence and species ofCryptosporidiumin camel populations across Australia are essential to better understand their potential for contamination of drinking water catchments.
Virulent T4 Acanthamoeba causing keratitis in a patient after swimming while wearing contact lenses in Southern Brazil
Authors: Fabres Laura Fuhrich , Maschio Vinicius José , Santos Denise Leal dos , Kwitko Sergio , Marinho Diane Ruschel , Araújo Bruno Schneider de , Locatelli Claudete Inês , Rott Marilise Brittes
Published: April 2018

Several strains of free-living amoebae belonging to the genusAcanthamoebacan cause a painful sight-threatening disease of the cornea known asAcanthamoebakeratitis (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 ofAcanthamoebastrains. 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.
Beyond ‘(non) native-speakerism’: Being or becoming a native-speaker teacher of English
Authors: Leonard Josie
Published: April 2018

The labelling of teachers of English as either ‘native’ or ‘non-native’ speakers in the field of English Language Teaching continues to promote ideals of ‘native-speakers’ that impact negatively on the teaching lives of those teachers using English as another language. In this paper, I explore constructs of ‘native-speakerism’ (Holliday, Adrian. 2015. Native-speakerism: Taking the Concept Forward and Achieving Cultural Belief. In Anne Swan, Pamela Aboshiha & Adrian. Hollliday (eds.),Encountering Native-speakerism: Global perspectives, 11–25. Basingstoke: Palgrave Macmillan) by examining them as networks or assemblages formed through interactions of people, technologies, discourses and other material objects integral to teaching and learning environments. Drawing on ‘Actor-network theory’, I analyse unique influences of ‘native’ and ‘non-native’ networks as experienced by individual teachers of English from different contexts. The data collected in this qualitative study shows how ‘native-speaker’ networks form and exert power to reinforce the ideal of ‘native-speaker’ teachers, and restrict the agency of those who are classed as ‘non-native’. By unravelling these networks, I challenge the notions on which they are constructed, and show how the categorising of teachers in this way undermines the legitimacy of those classed as ‘non-native’, and limits their professional development. I therefore argue that moving beyond these labels is an essential step for English Language Teaching to move forward as a profession.
Language Choice and Researcher’s Stance in a Multilingual Ethnographic Fieldwork
Authors: Gallego-Balsà Lídia
Published: April 2018

In bilingual settings language choice represents one of the main resources individuals have to construct their identities. According to Jaffe (2009), it is also a way in which individuals position themselves towards the languages of the sociolinguistic context. However, in the context of researching multilingual settings, very little attention has been paid to how the researcher’s language choice(s) interact with the participants’ language choices, and how the linguistic features that s/he activates contribute to the construction of the researcher’s identity and her/his stance towards the languages of the sociolinguistic matrix within which the research is embedded. This paper is based in the context of Catalonia and explores how the researcher projects and is ascribed a stance as a consequence of her own linguistic practices. First, it shows that these practices lead to the construction of relationships of (dis)affiliation between the researcher and the participants, which ultimately influence the type of data collected. Second, it suggests that language choice is an essential lens through which to look at the researcher’s positionality in multilingual settings and provide more transparent accounts of multilingual ethnographic research.
Doing ethnography among Latinos in London and Barcelona: languages, genders, and ethnicities
Authors: Corona Victor , Kelsall Sophie
Published: April 2018

Based on two ethnographic studies of people of Latin American descent in global cities, this article explores how language, gender, and ethnicity shape field relations, community membership, and data collection. It examines some of the implications of being positioned intersectionally as an outsider or insider of the community, and being sexualised by a male gatekeeper. It suggests that gender roles are a powerful aspect of conducting ethnographic research among Latinos, while pointing to the challenge of dealing with, and potentially contributing to, essentialising discourses in the field. It argues that the notion of ‘being Latino’ is imagined and constructed interactionally and contextually, in reaction to social pressures, as well as local and historical narratives.
The relationship between English proficiency and humour appreciation among English L1 users and Chinese L2 users of English
Authors: Chen Xuemei , Dewaele Jean-Marc
Published: April 2018

Humour appreciation involves split second detection and resolution of cultural and pragmatic incongruities. Second language (L2) users may need more time and effort to understand and appreciate L2 humour. Previous studies have mostly used decontextualized verbal jokes and reported a linear relationship between L2 proficiency and humour appreciation. The present study strives for more ecological validity by using audiovisual-based, multimodal humorous stimuli. A total of 272 Chinese L2 users of English and 94 English L1 users rated the funniness and the ease of understanding of two short video extracts and then completed an English vocabulary size test, LexTALE. The findings suggest that L2 users need to reach a certain threshold in L2 linguistic, pragmatic and sociocultural knowledge before a positive linear relationship emerges between proficiency and appreciation of multimodal humorous stimuli. Also, advanced L2 users demonstrated similarities with English L1 users in humour processing.
Biogas digestate – benefits and risks for soil fertility and crop quality – an evaluation of grain maize response
Authors: Przygocka-Cyna Katarzyna , Grzebisz Witold
Published: April 2018

The agricultural usability of biogas digestate solids (BDS) as a soil amendment depends upon its impact on soil fertility and the content of minerals in the edible part of the grown crop. This hypothesis was verified in a series of field experiments with maize conducted between 2014 and 2016 at Brody, Poland. The two-factorial experiment consisted of the DBS application method (broadcast and row) and its rate: 0, 0.8, 1.6, 3.2 t ha–1. The post-harvest analysis of soil fertility showed that BDS can, at least partly, replace mineral fertilizers. The supply of N-NO3 to maize as a growth driving factor was significantly limited by a shortage of iron, potassium and, to some extent, magnesium. As recorded in 2016, the shortage of available Fe resulted in a low/pool of N-NO3, thus significantly decreasing the yield of grain. The shortage of K supply to grain created a pathway for the accumulation of other elements, including heavy metals. The disadvantage of the N-NO3 pool increase, due to the DBS application, was concomitant with the enhanced intake of cadmium and lead, which consequently exceeded their permissible concentration limits in grain. These unfavorable results of biogas digestate impact on the quality of maize grain can be ameliorated by incorporating zinc into the biogas type of soil amendment and keeping a sufficiently high level of available potassium and iron. The shortage of K can be partly overcome by a better sodium supply, however, its accumulation in grain results in an enhanced accumulation of cadmium and lead.
Optimum Conversion of Major Ginsenoside Rb1 to Minor Ginsenoside Rg3(S) by Pulsed Electric Field-Assisted Acid Hydrolysis Treatment
Authors: Lu Chengwen , Yin Yongguang
Published: April 2018

Ginsenoside Rg3(S) is a primary bioactive component in ginseng, which has pharmacological effects and nutritional activities. In the present study, pulsed electric field (PEF)-assisted acid hydrolysis processing was used to convert major ginsenoside Rb1 to minor ginsenoside Rg3(S). The optimum parameters of PEF assisted acid hydrolysis were analyzed by response surface methodology (RSM). The optimum processing conditions were: electric field intensity, 20 kVcm−1; acid concentration, 0.25 mol/L; pulse number, 10. The conversion rate of ginsenoside Rg3(S) achieved 68.58%, in accordance to the predicted value. The structure of hydrolyzed product was confirmed by 13C-NMR. The results suggested that PEF-assisted acid hydrolysis significantly enhanced conversion rate of ginsenoside Rg3(S).
Relationships between diatoms and environmental variables in industrial water biotopes of Trzuskawica S.A. (Poland)
Authors: Malinowska-Gniewosz Agnieszka , Czerwik-Marcinkowska Joanna , Massalski Andrzej , Kubala-Kukuś Aldona , Majewska Urszula , Jankowski Michał
Published: April 2018

The heterogeneous nature and widespread anthropogenic impacts on industrial water biotopes in the Trzuskawica S.A., pose challenges to biomonitoring of this habitat. Generally, the concentration of trace elements in the industrial water biotopes reflects the anthropogenic impacts. With X-ray fluorescence method (TXRF) in waters 17 elements:P,S,Cl,K,Ca,Ti,Cr, Mn,Fe,Ni,Cu,Zn,Br,Rb, Sr, Ba, Pb were revealed. High amounts of Ca, from 300 ppm to 198 ppm in May and from 999 ppm to 231 ppm in September 2015, was determined. A total of 36 diatoms were found in two reservoirs and drainage ditch, but only three taxa: Cymatopleura radiosa, Navicula upsaliensis and Nitzschia angustata were present in all 7 sampling sites. These species are known to be tolerant to organic pollution, eutrophication, and also characteristic for limestone waters. The results of CVA showed that dintoms in the water reservoir stocked with fish were distinguished by highest species richness. The relationships between diatoms and environmental variables confirm the positive correlation with the currently functioning industrial plant (despite the increased water temperature and large content of trace elements). Our results suggest that, though heterogeneity in both diatoms and selected elements in industrial waters, diatoms can be useful indicators of habitat conditions.