Volume 60, Issue 2

Demands for effective vaccines to control parasitic diseases of humans and livestock have been recently exacerbated by the development of resistance of most pathogenic parasites to anti-parasitic drugs. Novel genomic and proteomic technologies have provided opportunities for the discovery and improvement of DNA vaccines which are relatively easy as well as cheap to fabricate and stable at room temperatures. However, their main limitation is rather poor immunogenicity, which makes it necessary to couple the antigens with adjuvant molecules. This paper review recent advances in the development of DNA vaccines to some pathogenic protozoa and helminths. Numerous studies were conducted over the past 14 years of 21 st century, employing various administration techniques, adjuvants and new immunogenic antigens to increase efficacy of DNA vaccines. Unfortunately, the results have not been rewarding. Further research is necessary using more extensive combinations of antigens; alternate delivery systems and more efficient adjuvants based on knowledge of the immunomodulatory capacities of parasitic protozoa and helminths.

During the studies on ecology of Trichoptera of anthropogenic water bodies we have unexpectedly discovered the parasitic larvae of water mites of the species Tiphys torris on the pupa of Triaenodes bicolor. This is the first documented case of the parasitism of water mites on the caddisfly pupa as well as the first ever record of the species which is regarded as a dipteran parasite on caddisflies. The situation is very untypical for preimaginal stages of caddisflies are used by phoretic and not parasitic water mite larvae. Parasitism has been confirmed in this case by the formation of stylostomes and enlarged sizes of the bodies of the larvae. This is probably the case of facultative parasitism in which the pupa has served as a substitute of the adult form of a caddisfly.

Only about 20 species of microsporidia have been described from mites. All except one species produce typical spores with a long polar filament and a polaroplast. This paper is the first study of an atypical microsporidium infection in a feather mite (Falculifer rostratus). The infection of the pigeon feather mite is restricted to the colon epithelium where it leads to hypertrophy of the concerned cells. During sporogony, a multinucleate plasmodial aggregate is formed within a sporont (endogenous sporogony resulting in a polysporophorous vesicle). The cisterns delimiting the single sporoblasts later form the spore walls. Sporogonial stages are in direct contact to the host cell cytoplasm. Merogonial stages were not present. Spores are tiny (3.6 μm × 2.6 μm), broad oval in form and monokaryotic. The spore wall of mature spores consists of a three-layered endospore and a thin, electron-dense, wavy exospore. The polar filament is anisofilar and completely coiled in 3-4 turns. In cross-sections, it has a star-like appearance because the electron-dense core forms rounded compartments of lucent material at its surface. In superficial sections, this results in a honeycomb-like pattern. A polaroplast is missing. The polar filament arises subapically at a polar sac that lacks an internal anchoring disk. These atypical spore structures clearly classify the species from the feather mite as a member of the order Chytridiopsida. It could not be clearly affiliated to one of the known genera, so we created a new genus, Acarispora, with the species A. falculifera.

Fasciolosis is an important parasitic zoonosis considered the most important helminth infection of ruminants in tropical countries. The aim of this study was to develop a PCR assay for the sensitive and specific detection of F. hepatica in formalin preserved sheep faeces. A 405-bp fragment of the cytochrome c oxidase subunit 1 gene of F. hepatica was amplified from stool samples of infected sheep. The PCR assay showed a detection limit of 20 pg of F. hepatica DNA. No cross-reactions were observed with samples containing coccidian oocysts or gastrointestinal nematodes eggs. Our PCR technique showed to be effective for specific detection of F. hepatica infections in sheep.

This study analyzed the relationship between intermittent preventive treatment with sulfadoxine-pyrimethamine (SP) (IPTp-SP), the rate of multiple resistant parasites and of submicroscopic gametocyte carriage among pregnant women at the beginning of IPTp implementation in Gabon (2005) and six years after (2011). The detection of pfdhfr and pfdhps gene mutations was performed by PCR-RFLP in Plasmodium (P.) falciparum positive samples collected from pregnant women in 2005 and 2011. Gametocytes carriage was detected by Pfs25mRNA amplification using QT-NASBA. Data were analyzed according to the time of collection (study period) and IPTp-SP doses. The proportion of isolates with at least a triple Pfdhfr mutation (n = 39/42, 92.9% versus 100%, n = 78/78)) and of those isolates with the S108N/C59R/N51I/S436A/A437G multiple mutation (17.9% versus 75.6%) significantly increased between 2005 and 2011 (p<0.01). Mutations I164L and A581G were not found, while higher proportions of 436 and 437 mutations were detected in 2011.A trend toward a higher frequency of isolates with five mutations was observed in women who received two SP doses (p<0.01). Pfs25mRNA was found in 6.8 % (n = 3/44) and 34.6% (n = 27/78) of the samples collected in 2005 and 2011 respectively (p<0.01). In 2011, 74.0% (n = 20/27) of women with detected submicroscopic gametocytes carried parasites with the S108N/C59R/N51/S436A/A437G multiple mutation. All the ten delivering women who received three IPTp-SP doses had a submicroscopic Plasmodium falciparum infection, but none had detected gametocytes. Following IPTp-SP implementation, an increase in the frequency of multiple mutant parasites and of submicroscopic gametocyte carriage was observed among pregnant women living in Gabon.

Based on light and scanning electron microscopical studies, one known and one previously undescribed Procamallanus (Spirocamallanus) species (Nematoda: Camallanidae) are described from specimens collected in the intestine of freshwater fishes in the Lower Congo River, Democratic Republic of Congo: P. (S.) daleneae (Boomker 1993) from Synodontis acanthomias Boulenger (Mochokidae, Siluriformes) (new host and geographical records) and P. (S.) parachannae sp. nov. from Parachanna insignis (Sauvage) (Channidae, Perciformes). The new species is most similar to P. (S.) serranochromis Moravec et Van As 2015, a parasite of African cichlids (Serranochromis spp.), differing from it mainly in the absence of any outgrowths on the female tail tip, a distinctly longer (546 μm) right spicule, a different length ratio of both spicules (1:2.6) and in the morphology of fourth-stage larvae. P. (S.) daleneae, a specific parasite of squeakers (Synodontis spp.), is redescribed; in contrast to the original description, the number of postanal papillae was found to be identical (6 pairs) with that of other related species.

Podocotyle bathyhelminthos n. sp. (Opecoelidae: Plagioporinae) is described from the cusk-eel, Luciobrotula corethromycter Cohen, 1964 (Ophidiiformes: Ophidiidae), collected at depths of 622-1,280 m in the northern Gulf of Mexico and Caribbean Sea off Colombia. The new species is distinguished by possessing a combination of the following features: an elongate oval body shape, a sucker width ratio of 1:1.4-1.9, deeply lobed and irregularly-shaped testes, a cirrus-sac that extends just in to the hindbody, a trilobed ovary and vitellarium that extend to the ventral sucker level. Several unique features in P. bathyhelminthos n. sp. were not present in all, or almost all, recognized species of Podocotyle including a conspicuous deep cleft at the posterior end of the worm, a small transverse ridge on the ventral surface immediately anterior to the ventral sucker, uterine loops extending ventral to the caeca and, at times, lateral to the caeca, a thick-walled metraterm extending 1/3 to 1/2 the length of the cirrus-sac and P. bathyhelminthos n. sp. parasitizes a deep water piscine host. Podocotyle etheostomae Aliff, 1973 is declared a nomen nudum. A brief discussion of Podocotyle Dujardin, 1845 in deep waters is presented, and a gastropod, caridean shrimp and cusk-eel are hypothesized as hosts in the life cycle of P. bathyhelminthos n. sp. in the deep sea.

Plasmodium vivax is the most common cause of malaria worldwide as well as southeastern Turkey. After the implementation of a successful national elimination program that the local malaria cases were not reported in 2011, malaria returned to county of Savur located in southeastern Turkey in summer of 2012. The present study aimed to determine the prevalent P. vivax genotypes isolated from southeastern Turkey. Genetic polymorphism in P. vivax CSP gene was analyzed by PCR-RFLP to assess the ratio of VK210 and VK247 types. Blood samples were obtained from 15 patients who lived in southeastern between 2005-2006. According to the results, VK210 type was detected in 10 samples (66.6%), VK247 type was observed in three samples (20%). Remaining two samples showed mixed infection (13.3%). The results of the present study first time showed the ratio of P. vivax genotypes in southeastern Turkey before the elimination in 2011. The results of the present study will be enable researchers to compare the new isolates with the previously detected ones and design new treatment and/elimination strategies.

A new feather mite species Dubininia microhieracis sp. n. (Analgoidea: Xolalgidae) is described from the Black-thighed Falconet Microhierax fringillarius (Drapiez) (Falconiformes: Falconidae) from Java. This is the second species of the genus Dubininia found on falconiform hosts; most known species of this genus are associated with parrots (Psittaciformes).

The first data on the parasites of the giant trahira, Hoplias aimara (Characiformes: Erythrinidae), a little known, but popular fish for sport angling, are provided. In H. aimara from the Xingu River in Brazil, two new species of gill monogeneans of the family Dactylogyridae were found and their description is provided herein. Urocleidoides aimarai n. sp. differs from all 18 congeners by the presence of a large accessory piece of the male copulatory organ and based by morphology of anchors. Urocleidoides xinguensis n. sp. is distinguished by unique combination of copulatory complex and haptoral structures.

Cosmocerca microhylae sp. nov., recovered from the rectum of a red narrow-mouthed frog, Microhyla rubra (Jerdon, 1854) (Anura: Microhylidae), collected from Bolpur in the Birbhum district of West Bengal, India, is described and figured. This species is similar to C. acanthurum, C. banyulensis, C. cruzi, C. japonica, C. kalesari, C. novaeguineae, C. ornata, C. paraguayensis, C. parva, C. podicipinus and C. travassosi in having 5 pairs of plectanes supporting preanal papillae but differs from these species by smaller size, absence of somatic papillae in females and having only one pair of adanal papillae in males and one pair postanal papillae in females. Cosmocerca microhylae sp. nov. represents 27th species assigned to the genus, and 4 th species from India.

Hepatozoon species are the most common hemoparasites of snakes. In this study, Hepatozoon parasites were examined for the first time in king rat snakes (Elaphe carinata) from Shanghai, China. All 10 snakes were found to be infected with Hepatozoon gamonts. The gamonts were folded back in a hook-wise fashion for about 3 μm at one end. Parasitemia levels ranged from 4-43 infected erythrocytes per 1,000 examined. The gamonts changed the morphology of the parasitized erythrocytes. Although the gamonts showed some distinct variations in both the parasite and its nucleus, phylogenetic analysis indicated that all the E. carinata in this study formed a monophyletic group, and were distinct from all other published Hepatozoon species. A new species, Hepatozoon chinensis, was proposed based on the molecular and morphologic evidence.

Neospora caninum is considered one of the major causes of abortion in cattle. The aim of this study was to examine and quantify the extent of the infection in cattle in a representative region of Argentina (La Pampa, province). An average sample size of 36 sera per herd was selected from 97 beef and 24 dairy herds. A total of 4334 serum samples were tested for specific anti- Neospora caninum IgG using an indirect-ELISA and 302 seropositive-ELISA sera were re-examined using an Avidity-ELISA procedure for N.caninum. The overall estimated seroprevalence for N.caninun was 9.6% (95%CI: 8.7%; 10.5%). Levels of seroprevalence were significantly different in beef 7.0% and dairy 20.3% cattle. Disease distribution seems to be associated with climatic conditions as well as the management system. Cows in the east and central regions were at a 4.5-fold and 2.0-fold higher risk, respectively, of being N. caninum seropositive compared with cows in west region. Levels of recent infection were evaluated through an avidity ELISA in seropositive animals, being registered a 0.56% and a 1.71% of recent infection in beef and dairy cattle respectively (p = 0.006). The results revealed that dairy cows had 3.1(95%CI: 1.4; 7.0) higher risk of contracting Neoporosis through horizontal transmission than beef cows. A relationship between Brucella abortus and N. caninum seroprevalence was also observed. The risk of being N. caninum seropositive was two times higher where Brucellosis seroprevalence was >3.5%. These results reveal the distribution of N. caninum infection in the cattle population in La Pampa, Argentina

The members of the Anopheles maculipennis complex have been incriminated for the transmission of the malaria in Europe, which was endemic until the middle of the century. The global warming and the intensification of the intercontinental travel constitute a risk of the re-emergence of the malaria in Europe, given the presence of the Anopheles vectors. The study has attempted the identification by using the PCR (Polymerase Chain Reaction) of the members of the Anopheles maculipennis complex from the North-eastern area of Romania from the city of Iaşi. In total there have been identified by using the PCR amplifying the ITS2 sequence of the ribosomal DNA, 217 specimens belonging to the complex of A. maculipennis among which: 58 A. atroparvus, 18 A. melanoon, 2 A. labranchiae, 52 A. maculipennis and 87 A. messeae. The ITS2 sequences of the ribosomal DNA have been compared to those of the species belonging to the A. maculipennis available in GenBank. The Species A. labranchiae is reported for the first time in Romania, being identified in the larval stage IV. The adaptation of a new species to the climatic conditions present in the North-eastern Romania, confirms the phenomenon of global warming and also the intensification of the travelling. As a result of the analysis of the A. labranchiae sequence, this one corresponds to the extent of 96% to the species from Italy, registered in GenBank, given the fact that a high number of the inhabitants of the municipality of Iaşi are working in this country.

We identified the ectoparasites and helminth fauna of yellow-legged gulls (Larus michahellis michahellis), breeding near to a solid waste landfill, and compared infection levels with those of other yellow-legged gull colonies. Moreover, we analysed correlations between parasites and sex and body condition of yellow-legged gulls, co-infections and the helminth community structure in order to propose the role of this species as reservoir of certain parasites. We also discuss the potential transmission of parasites between the yellow-legged gull and the endangered Audouin’s gull, because interactions between these two species, such as kleptoparasitism and predation, occur frequently around colonies. The following species were recorded: Ornithodorus capensis (Arthropoda); Cosmocephalus obvelatus, Paracuaria adunca, Eucoleus contortus, Tetrameres skrjabini and Contracaecum sp. (Nematoda); Tetrabothrius cylindraceus (Cestoda); Acanthotrema armata, Cardiocephaloides longicollis and Ornithobilharzia intermedia (Digenea). Tetrabothrius cylindraceus, A. armata and O. capensis are new parasite records for this host. The dependence of yellow-legged-gulls on fishery discards is supported by the dominance of parasites transmitted through marine intermediate hosts with interest to fisheries in the study area. However, the shift in diet from natural resources to food derived from human activities seems not to affect the parasitic fauna of yellow-legged gull. Besides of direct physical contact between individuals in nesting and resting habitats, the high availability of fishery discards could increase the risk of Audouin’s gulls to be infected by common parasites of yellow-legged gull.

Phyllodistomum spinopapillatum sp. nov. is described from the urinary bladder of the Oaxaca killifish, Profundulus balsanus Ahl (Profundulidae) in Rio Pueblo Viejo and Rio Santa Cruz, Oaxaca, southwestern Mexico. The new species is described based on evidence gathered from morphology, scanning electron microscopy (SEM), and partial sequences of the 28S rRNA gene. Diagnostic characters of the new species of Phyllodistomum Braun 1899 are the presence of spines on the entire body surface and having a ventral sucker almost half the size of oral sucker. The new species possess a large number of dome-like papillae on the body surface with scattered distribution along the hindbody, and these papillae are characteristically spinulated. Phyllodistomum spinopapillatum sp. nov. most closely resembles P. inecoli Razo-Mendivil, Perez-Ponce de Leon and Rubio- Godoy, 2013, a parasite of the twospot livebearer Pseudoxiphophorus bimaculatus (Heckel) from Veracruz, in the Atlantic slope of Mexico. In addition to the new species, specimens of P. inecoli were also found parasitising the urinary bladder of cyprinodontiforms such as the Mexican molly Poecilia sphenops Valencienes in a pond at Santa Maria Coyotepec, and in Profundulus sp. in Rio Templo, both in Oaxaca, and in the Porthole livebearer Poeciliopsis gracilis (Heckel) in Rio San Juan, as well as in Profundulus punctatus (Gunter) from Rio Nueva Francia, both in Chiapas. The distribution and host range of P. inecoli is extended to freshwaters of the Pacific slope of Mexico, and to other cyprynodontiforms.

A new species of Coronocestus Caira, Marques, Jensen, Kuchta and Ivanov, 2013 is described from Iago omanensis (Norman) from the Gulf of Oman. Coronocestus ehsanentezarii sp. nov. differs from C. musteli (Pintner, 1889) Caira, Marques, Jensen, Kuchta and Ivanov, 2013 in that its ovary is H- rather than U-shape. The new species is easily distinguished from C. diamanti (Ivanov and Lipshitz, 2006) Caira, Marques, Jensen, Kuchta and Ivanov, 2013 by the number of spines per column on the cephalic peduncle (24-36 vs 95-118). It differs from C. hormozganiense (Haseli, Malek, Palm and Ivanov, 2012) Caira, Marques, Jensen, Kuchta and Ivanov, 2013 based on a greater number of spines per column on the cephalic peduncle (24-36 vs 18-21). The new species differs from C. notoguidoi (Ivanov 1997) Caira, Marques, Jensen, Kuchta and Ivanov, 2013 in that of its scolex is craspedote rather than acraspedote. Unlike C. coronatum (Robinson 1959) Caira, Marques, Jensen, Kuchta and Ivanov, 2013, C. ehsanentezarii sp. nov. possesses 29-35 rather than 20 apical hooks in each dorso-ventral group. The new species possesses lateral hooklets with two rows (a and b designations) in each cluster. This character had been presented earlier only for Andocadoncum Abbott and Caira, 2014. Furthermore, a new locality record is presented for C. diamanti from Iago omanensis in the Gulf of Oman. Thus, I. omanensis certainly hosts two diphyllidean species simultaneously in the Gulf of Oman. The generic diagnosis of Coronocestus is also revised to include new data.

In Colombia it is estimated that about 900,000 persons are infected with T. cruzi. There are 25 triatomine species and 5 of them have been reported infected with T. cruzi in the Colombian Caribbean region. In order to obtain more information about the triatomine populations in this region, 89 wild triatomines were collected from four Colombian Departments. The most frequent specie collected was Rhodnius pallescens (65%), followed by Rhodnius prolixus (20%), Panstrongylus geniculatus (10.1%) and Triatoma dimidiata (1%), found in Bolivar, Córdoba, Atlántico/Sucre, and Bolívar Departments, respectively. The majority of triatomines (95.5%) were captured in the arboreal ecotope and 76.4% were found infected with T. cruzi. Interestingly, some of these triatomine species were captured in Departments in which they had not previously been reported and also new finding of triatomine species infected with T. cruzi. These results are relevant, because they can be consequence of a continued geographical expansion of this parasite, not only in the Colombian Caribbean region, but even in all Latin America. The information presented here will contribute in the surveillance and control strategies of the vectors infected with T. cruzi that circulate in four department of Colombian Caribbean region in order to interrupt the transmission to human dwelling.

A sizeable Indian equine population is considered to be pre-immune carrier of Theileria equi infection. In this study we confirmed the presence of T. equi specific DNA in Hyalomma anatolicum ticks which were infested on sero-positive horses. Fifty two Indigenous horses were randomly selected from endemic areas and their blood and tick samples were collected. Tick salivary glands and blood samples were processed for separation of DNA and serum, respectively. Serum samples were analyzed by EMA-2 ELISA and nine horses were found positive for T. equi specific antibodies. Species-specific primers were designed from EMA-2 gene of T. equi, so as to amplify 398 bp fragment in PCR. The gene fragment was amplified in PCR on the DNA samples (from blood) from these nine sero-positive horses. Corresponding six tick’s DNA samples collected from these nine seropositive animals were observed positive in PCR. Further, qPCR assay demonstrated presence of T. equi DNA in infected tick’s salivary glands, which was also confirmed by microscopic examination of infected acinar. This study concluded that Hyalomma anatolicum ticks infested on T. equi seropositive horses have sporozoite developmental stage in their salivary glands, which is an evidence for transmitting potential of these tick among Indian horse population.

A new microsporidium, Nosema sp. PM-1, was first isolated from Papilio machaon Linnaeus. The spore shape of the PM-1 isolate was a long oval with an average size of 3.22 μm × 1.96 μm. Ultrastructure observation showed that PM-1 had a typical Nosema common diplokaryotic nuclei structure with 10-13 polar filament coils, spore wall, plasma membrane, and anchoring disk. The complete rRNA gene sequences were obtained by polymerase chain reaction amplification and each rRNA unit was arrayed as follows: 5′-LSU (2497 bp)-ITS (179 bp)-SSU (1232 bp)-IGS (278 bp)-5S (115 bp)-3′, which was the same as typical Nosema. The phylogenetic trees of rRNA, DNA-directed RNA polymerase II subunit, and tubulin genes all show that PM-1 was a sister to the clade comprising Nosema bombycis, Nosema spodopterae, and Nosema sp. PX1. The spore morphology, ultrastructure, and complete rRNA structure indicate that this isolate assigned to the ˝true˝ Nosema group, can parasitized in Papilio machaon Linnaeus, which provides a wider host range for Nosema.

The probability of infection with fungi, as well as parasitic nematodes or arthropods may increase in overcrowded population of animals and human. The widespread overuse of drugs and immunosuppressants for veterinary or medical treatment create an opportunity for many pathogenic species. The aim of the review is to present the common molecular characteristics of such pathogens as fungi and nematodes and other chitin bearing animals, which may both activate and downregulate the immune response of the host. Although these pathogens are evolutionary distinct and distant, they may provoke similar immune mechanisms. The role of chitin in these phenomena will be reviewed, highlighting the immune reactions that may be induced in mammals by this natural polymer.

We aimed to study the effectiveness of hyperbaric oxygen therapy (HOT) (100% oxygen at 2 ATA for 70 minutes each session for 20 consecutive days) on BALB/c male mice infected with Leishmania major. Fifty-one mice were assigned to six groups. Group 1 was treated with HOT from 1 day after the inoculation. In Groups 2-5, treatment began when the cutaneous lesions appeared. Group 2 received HOT only, Group 3 received topical therapy with Leshcutan only, Groups 4 and 5 received a combination of HOT and Leshcutan for 5 and 10 days respectively, and Group 6 did not receive any treatment (control group). When comparing the control group with Group 1, treatment with HOT in Group 1 did not significantly affect the time of the appearance of the lesions. In contrast, mice treated with Leshcutan demonstrated a significant difference in lesion size and spleen dimensions as compared to the rest of the mice (p<0.001). The results show that HOT treatment has no positive effect on the course of Leishmaniasis in a BALB/c mice model infected with Leishmania major. Further studies are needed with a mouse model closer to humans and with different HOT protocols.

The European rabbit Oryctolagus cuniculus is a keystone species from the Iberian Peninsula where viral diseases have played a prominent role in regulating their populations. Coccidiosis, a parasitic disease caused primarily by Eimeria spp., is also thought to have important negative effects. However, few studies have investigated the impact of coccidia on wild European rabbit populations on the Iberian Peninsula. Here we estimate coccidian prevalence in rabbit faecal samples collected along transects established in two ecological regions. Six Eimeria species, with different pathogenicity, were identified (E. coecicola, E. perforans, E media, E. magna, E. irresidua and E. flavescens). Species diversity varied significantly between regions although mean oocyst excretion levels were generally low in both areas (57.61 s.d.±78.07 and 17.03 s.d.±27.72, oocyst per gram of rabbit faeces). This study is the first to describe the composition of the Eimeria spp. assemblage for wild rabbit populations on the Iberian Peninsula and provides fundamental information for future studies on the potential interaction of viral and parasitic diseases.

Angiostrongylus vasorum (Nematoda: Angiostrongylidae) infection was detected at post-mortem examination in the pulmonary arteries and hearts of 34/102 (33,3%) of red foxes (Vulpes vulpes) from the Campania Region in southern Italy. Pathological changes consisted of granulomatous interstitial pneumonia caused by larvae and intravascular pulmonary adult nematodes. These changes confirm that angiostrongylosis infection in red foxes has a mainly chronic course, in which the infected host may disperse parasite larvae in the environment over its lifetime. Results suggest that the life cycle of A. vasorum is well established in the red fox in the Campania Region representing a potential infection risk for dogs.