Volume 60, Issue 1

The paper presents an overview of the use of natural therapeutic agents in combating parasitic diseases. Nowadays there is increasing demand for proven plant therapies, which often are found to be more effective than synthetic pharmaceuticals in chronic diseases. In many cases herbal preparations perfectly supplement the conventional treatment and at the same time do not cause side effects. On the pharmaceutical market there are many drugs of plant origin which have been applied in the treatment of parasitic diseases. However, researchers are still looking for new plants, or specific substances isolated from them, which can be used in therapy. In this paper, drugs of plant origin used in the treatment of amoebiasis, giardiasis, malaria, leishmaniasis, trypanosomiasis and acanthamoebiasis are described.

Fish is an important food source for an estimated 300,000 people inhabiting the shores of Lake Turkana, Kenya. Despite its large size (7560 km2) and importance, the lake remains one of the least studied in the Great Lakes Region of Africa. This study describes the parasite community of the elongate tigerfish Hydrocynus forskahlii (Cuvier, 1819) and the redbelly tilapia Tilapia zillii (Gervais, 1848). A total of 87 individuals (43 H. forskahlii and 44 T. zillii) were dissected and examined for parasites. Two taxa infected H. forskahlii, the dominant one being an anisakid nematode Contracaecum sp. (L3) (P = 83.7%, mean intensity (MI) = 46.0, abundance (A) = 38.5). Twelve parasite taxa were recovered from T. zillii, with metacestodes of the gryporhynchid cestode Amirthalingamia macracantha being dominant (P = 79.5, MI = 10.3, A = 8.2). This was the first report of a mixture of merocercoids and plerocercoids in the same host. Fish size was positively correlated with the major parasite infection levels, but, the prevalence of Contracaecum sp. in H. forskahlii was negatively correlated with size, probably reflecting ontogenetic shifts in habitats and diets with age. Fish sex was not a significant influencing factor, with the exception of Contracaecum sp. in H. forskahlii, where prevalence differed significantly between sexes (p<0.05), with the females having a higher prevalence than the males. We conclude that H. forskahlii had a poor parasitic community but that the infection levels with Contracaecum sp. were high. T. zillii had a rich parasite fauna, although, most parasites occurred at low intensities.

Although vector-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on these conditions in Costa Rica. In PCRs of blood from dogs in Costa Rica, we did not detect DNAs of Rickettsia (R.) felis and Coxiella (C.) burnetii but we did find evidence of infection with Dirofilaria (D.) immitis (9/40, 22.5%), Hepatozoon (H.) canis (15/40, 37.5%), Babesia spp. (10/40, 25%; 2 with B. gibsoni and 8 with B. vogeli), Anaplasma (A.) platys (3/40, 7.5%) and Ehrlichia (E.) canis (20/40, 50%). Nine dogs (22.5%) were free of any vector-borne pathogens while 14 (35%) were infected with a single pathogen, 11 (27.5%) with two, 4 (10%) with three, 1 (2.5%) with four, and 1 (2.5%) with five pathogens. Dogs in Costa Rica are commonly infected with vector-borne agents

Cryptosporidium is a genus of zoonotic pathogens transmissible from a variety of animals to humans and is a considerable public health concern. It is a significant cause of diarrheal disease in developing and industrialized nations. Cryptosporidium parvum and Cryptosporidium hominis are the main agents of cryptosporidiosis in humans. In this study we identified the genotypes of the Cryptosporidium isolates from clinical samples from diarrheic children using polymerase chain reaction (PCR) amplification and restriction fragment length polymorphism (RFLP) analyses of the TRAP-C2 gene (Thrompodin Related Adhesive Protein). A total of 430 fecal specimens from 1 to 14 years children were collected from inpatient and outpatient clinics of Benha University, Educational and Children Specialized Hospitals, Benha, Qalubyia, and were microscopically examined for Cryptosporidium spp. All infected samples were also analyzed using nested PCR. A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the (266-366 bp) of TRAP-C2 gene was also used to detect and identify Cryptosporidium spp. in PCR- positive samples. The results showed that 50 (11.63%) of the specimens were positive for Cryptosporidium spp. Genomic amplification and restriction digestion of the PCR products by BstETI, Hae III for TRAP-C2 gene restriction enzymes revealed that 82% (41/50) had C. parvum, 12% (6/50) had C. hominis, and three (3/50) samples (6%) had mixed infections. In conclusion, elevated prevalence of C. parvum, suggesting animal-human (zoonotic) transmission and further investigations are required to determine the subgenotypes of C. parvum to clarify the mode of transmission in order to improve the control measures.

Larvae of Metastrongylus spp. lungworms infect wild boar (Sus scrofa) definitive hosts through earthworms (Lumbricidae). We compared the abundance and Metastrongylus spp. larval infection measures of earthworms between two areas (both in Zselic, Hungary, 2012) characterized by markedly different wild boar population densities. Estimated wild boar density was 0.03 animal/ha in free range area and 1.03 animal/ha in enclosure. The mean abundance of earthworm populations (mostly Allolobophora, Aporrectodea, and Lumbricus spp.) was assessed by analysing 140-140 soil samples. The assesment of Metastrongylus spp. larval infection measures was based on cca 100-100 earthworms derived from the two areas. The abundance of earthworms and their Metastrongylus spp. larval infection measures (prevalence and mean intensity) were significantly lower in the free range than in the enclosure. Furthermore, using a finer scale within the enclosure, we compared wild boar feeding sites (n = 30) to other sites (n = 75). Earthworm populations were significantly more abundant and carried significantly more prevalent and more abundant larval lungworm infections at the former sites. These results suggest that high wild boar density and forage supplementation in enclosures increase both the abundance and the larval Metastrongylus infections of earthworms

We studied the long-term infection of Bithynia troscheli (Paasch, 1842) snails with trematodes and estimated the influence of trematode parthenites on the individual fecundity of female snails from the Kargat River (Chany Lake, Russia). The prevalence of B. troscheli females infected by trematode parthenites varied from 7.12% to 17.35% in different years. Eleven redioid species from 5 families and 9 sporocystoid species from 5 families of trematodes were found during this investigation. Snails’ fecundity was analysed in relation to the type of infection (redioid or sporocystoid species). Fecund females of B. troscheli were uninfected or they had pre-patent infections from 5 families of trematodes (Psilostomidae, Notocotylidae, Prosthogonimidae, Lecithodendriidae, and Cyathocotylidae). 89.9% of infected B. troscheli females were infertile. Moreover, 13.57% and 1.29% egg capsules (of infected and uninfected females, respectively) were without embryos (χ2 = 323.24, p<0.001). The results of the two-way analysis of variability confirmed that trematode parthenites influenced significantly the individual fecundity of B. troscheli. The age of B. troscheli females alone did not alter the individual fecundity, however age in combination with infection by tremathode parthenites influenced significantly the number of normal egg capsules. We also found that under unfavorable environmental conditions the proportion of fertile females increased by 23.7% among uninfected and by 219% among infected females.

Microsporidiosis is considered to be emerging opportunistic infection in immunocompromised individuals worldwide. The aim of this study was to identify the specific serum antibodies to intestinal microsporidia Encephalitozoon cuniculi and Encephalitozoon intestinalis in women with Human Papillomavirus HPV and without HPV by the indirect immunofluorescence (IFA). From total number of 669 examined women, 225 were HPV positive and 444 women HPV negative. Overall the study comprised of 10.8% women with positive result for presence of E. cuniculi antibodies. In group 1 (HPV-positive women) it was more than 28% and in group 2 (HPV-negative women) it was less than 2% (p<0.001). E. intestinalis infection was found in total of 4.48% women, in group 1 it was present in less than 6% and in group 2 in less than 4% of women.

Apyrase encoding metal-ions activated plasma membrane protease is present in animal and plant tissues. This enzyme can hydrolyze ADP and ATP pyrophosphate bond, resulting in AMP and free phosphate groups, and plays an important role for insects and parasites to evade host immune system. However localization and function of apyrase in the canine hookworm, Ancylostoma caninum, remains unknown. To analyze apyrase gene in A. caninum (a eukaryotic parasitic hookworm), a pair of primers was designed according to the previous EST data. The full-length cDNA of apyrase gene was amplified from A. caninum by RT-PCR. The partial cDNA of apyrase encodes 249 amino acid protein was expressed in Escherechia coli. The recombinant protein was induced to express under proper conditions and the molecular size was as expected. The recombinant protein was purified. The transcripts of apyrase in different stages of A. caninum were analyzed by the Real-time PCR assay, and Immuno-localization assays were used to research the protein expression in different stages of A. caninum

An isolate of Trypanosoma cruzi obtained from P. megistus captured in the peridomicile area of a home in Santo Antonio do Jardim city in the State of Sao Paulo, denominated T. cruzi Mogi, was characterized biologically and molecularly. The RFLP analysis of the D7 divergent domain in the 24Sα rDNA and of the mini-exon positioned the T. cruzi isolate within the TcI group. Phylogenetic analysis performed with the trypanosomatid barcode confirmed that the isolate belongs to the TcI group, with high homology to the 3014 c1 T.cruzi strain. The biological characterization of the isolate in rats showed a prepatent period of about 8 days, low parasitemia and tropism for cardiac, skeletal and colonic muscles. In Swiss mice the T. cruzi Mogi isolate showed a prepatent period of about 22 days, intermittent parasitemia in some animals, and tropism for cardiac and colonic muscles. Despite the inherent difficulty of identifying correlations amongst the molecular and biological characteristics of different T. cruzi groups, the tropism for colonic muscle demonstrated by T. cruzi Mogi represented a peculiarity of this isolate within the TcI group.

A Gyrodactylus parasite infected juveniles of brown trout (Salmo trutta L.) in a hatchery in Lake Janisjarvi, Russian Karelia. Molecular identification by ITS1-5.8S-ITS2 of ribosomal DNA indicated that the infection was caused by a non-segregating hybrid clone between unknown Gyrodactylus species and Gyrodactylus teuchis Lautraite, Blanc, Thiery, Daniel & Vigneulle, 1999, described from brown trout collected in France. The mitochondrial CO1 was sequenced from the hybrid, but it is not available from pure G. teuchis from type locality. The mitochondrial DNA was an independent clade among the wageneri group parasites, differing from the nearest relative G. derjavinoides by 19%. Morphometric measurements of the Janisjarvi parasite were compared with separate (host specific) G. salaris strains from farmed rainbow trout and salmon in Lake Onega, and with available data on G. teuchis from Western Europe, Austria and Poland. All isolates were distinguishable by morphometry, and the measurements are a useful primary diagnostic tool for the salmonid parasites in Karelian great lakes and fish farms. Pure G. teuchis is not found on Russian territory, and the other parent of the hybrid has not been discovered yet in any other country. A mini review of Gyrodactylus on salmonids in the Russian Karelia is presented.

Mitochondrial DNA was explored to study phylogeography of the nematode parasite Heligmosomum mixtum and elucidate molecular evolution pattern of cytochrome b gene. The size of cyt b gene ranged from 511 bp to 591 bp and the average of GC contents was 28.9%. The overall transition/transversion ratio R was 5.773 indicating that the transitions are more frequent than transversion. The aligned sequences allowed identifying 54 mtDNA haplotypes among the 119 examined individuals. The genetic divergence registered among the populations of H. mixtum was low (0.3% to 1.5%). Neighbor-joining and maximum Likelihood trees evidenced a huge polytomy and unstructured phylogeographic pattern among the studied populations. The demographic analyses tend to evidence a recent and rapid expansion of H. mixtum. Our results imply a positive selection and the genetic hitchhiking effect is unlikely. Parameters performed supported scenario of sweep selection and recent expansion of H.mixtum populations. Both positive selection and demographic histories have jointly contributed to the observed patterns of nucleotide diversity and haplotypes structure. The comparison of the phylogeographical pattern of H. mixtum with the one of its most common rodent host M. glareolus, confirmed a strong incongruence between the two species. These results strongly suggest that the parasite would not be specific to M. glareolus and that it would switch easily from one rodent species to another. The mitochondrial diversity seems to be unstructured with any biogeographic repartition of the variability and that the genetic structure of H. mixtum is probably associated with weak host specificity.

The purpose of this paper was to present the current data on the risk of toxocarosis in humans in Poland and to give an overview of the clinical and diagnostic aspects of the disease. The number of reported clinical cases of Toxocara infection in children in Poland in medical literature has increased recently. The results of field surveys aimed to evaluate the soil contamination with geohelminth eggs conducted during the last few years showed that Toxocara is the most common zoonotic agent in urban public sites and in rural settlements. The questionnaire revealed rural inhabitants’ low awareness of zoonotic parasite threats to humans. In particular parents should be advised as to what proper preventive measures to undertake to eliminate the toxocarosis risk factors for children in rural environment. Prevention of initial environment contamination with Toxocara canis and T. cati eggs, which includes proper treatment regimes to eliminate patent infections in dogs and cats and preventing pets from defecating in public areas and private households is vital. To provide the public with suitably presented information as well as pet owners with uniform recommendations, a close collaboration between veterinary and public health professionals is crucial.

Multidrug resistance Plasmodium falciparum is the major health problem in Thailand. Discovery and development of new antimalarial drugs with novel modes of action is urgently required. The aim of the present study was to investigate the antimalarial interaction of 9-hydroxycalabaxanthone and α-mangostin with the standard antimalarial drugs mefloquine and artesunate in chloroquine sensitive (3D7) and chloroquine resistant (K1) P. falciparum clones in vitro. Median (range) IC50 (drug concentration which produces 50% parasite growth inhibition) values of the 9-hydroxycalabaxanthone, α-mangostin, artesunate and mefloquine for 3D7 vs K1 clones were 1.5 (0.9-2.1) vs 1.2 (1.1-1.6) μM, 17.9 (15.7.0-20.0) vs 9.7 (6.0-14.0) μM, 1.0 (0.4-3.0) vs 1.7 (1.0-2.5) nM, and 13.3 (11.1-13.3) vs 7.1 (6.7-12.2) nM, respectively. Analysis of isobologram and combination index (CI) of 9-hydroxycalabaxanthone with artesunate or mefloquine showed synergistic and indifference antimalarial interaction, respectively. α-mangostin-artesunate combination exhibited a slight antagonistic effect of antimalarial interaction, whereas α-mangostin and mefloquine combination showed indifference interaction in both clones. The combination of 9-hydroxycalabaxanthone with α-mangostin showed the synergistic antimalarial interaction in both clones

Oswaldofilaria acanthosauri sp. nov. from the body cavity of the Cardamom Mountain horned agamid, Acanthosaura cardamomensis (Sauria: Agamidae), collected in Pursat Province, Cambodia is described. Of the 14 species assigned to Oswaldofilaria, O. acanthosauri sp. nov. is most similar to those species with spicular ratio of less than 2, namely, O. brevicaudata and O. chlamydosauri. Oswaldofilaria acanthosauri sp. nov. is easily separated from these 2 species in that O. brevicaudata is a South American species and in O. chlamydosauri the distal ends of the spicules are pointed not blunt. Mature individuals of 2 additional species of Nematoda, Meteterakis singaporensis and Orneoascaris sandoshami, as well as larvae assignable to Ascariidae were found. Acanthosaura cardamomensis represents a new host record for Meteterakis singaporensis, Orneoascaris sandoshami and Ascariidae (larvae).

Blastocystis is a common parasite and regarded as one of the etiologic agents of irritable bowel syndrome, colitis and chronic diarrhea. Our study was undertaken in order to identify different subtypes of Blastocystis isolated in patients with irritable bowel syndrome and colitis, as well as with chronic diarrhea and to evaluate their pathogenic potential. Seventy-three subjects (10 asymptomatic infected subjects, 49 subjects harboring Blastocystis or associated with other etiologic agents like bacteria, yeasts, protozoa, helminthes and 14 subjects with unknown etiologic agents) were investigated by in vitro parasitological and bacteriological stool samples followed by PCR subtyping of Blastocystis using STS primers, immunological markers (total serum IgA and IgE antibody levels), Helicobacter pylori antigen rapid test and fecal occult blood test. Also, among 49 subjects, there were 12 subjects harboring Blastocystis as the single etiologic agent. Subtyping proved that only three subtypes of Blastocystis were identified as following: subtype II (66.66%) in single infected subjects, subtype I (16.66%) in mixed infection with subtype II and subtype IV (8.33%) in single infected subjects. Total serum IgA and IgE antibody levels were in normal range. Subtype II was the most frequent subtype followed by subtype I and subtype IV of Blastocystis isolates in patients with irritable bowel syndrome, colitis, and chronic diarrhea as well as in asymptomatic infected group. Our results suggest that the severity of clinical manifestations depend on factors involving the host and possible parasitic density and not necessarily by isolated subtype.

The microphallids Microphallus szidati and Levinseniella cruzi parasitized at least three crustacean species as second intermediate host in Mar Chiquita coastal lagoon: the crabs Neohelice granulata and Cyrtograpsus angulatus and the grass shrimp Palaemonetes argentinus. Palaemonetes argentinus acts as host for both metacercariae in brackish and freshwater environments; however the seasonal variations of these digeneans in freshwater habitats remain unexplored. The aim of the present study was to analyze the seasonal dynamic of the larval digenean populations parasitizing P. argentinus in a lentic environment. From the total sample of 600 P. argentinus collected seasonally, 28.370 and 194 metacercariae of M. szidati and L. cruzi, respectively, were recovered. Microphallus szidati was characterized by high prevalence, 100% in all seasons. In contrast, L. cruzi presented low prevalence values, which increased towards summer. The two species of digeneans exhibit seasonal fluctuations in intensity of infection, which was related to host size. Temporal variation of larval digenean assemblage in the crustacean host is probably associated with the presence of the other hosts included in their life cycles. The high values of prevalence found for M. szidati in Nahuel Rucá lagoon suggest that P. argentinus could favor the contact between the larval digenean and the definitive hosts, establishing the presence of M. szidati also in freshwater environments and, thus, increasing the presence of the parasite in different areas of Mar Chiquita coastal lagoon.

Two new Dactylogyrus spp., D. angorae and D. turcicus have been described from the gills of the endemic cyprinid fishes, Capoeta angorae and C. mauricii from the Anatolian Region of Turkey. The shape of the anchors, hooks and the copulation organ of both species differ from those of the Dactylogyrus spp. found on other Capoeta species known from the Ponto-Caspian and Tigris-Euphrates regions.

Toxoplasma gondii is an obligate intracellular apicomplexan parasite that causes abortion and reproductive disorder in domestic animals. T. gondii is a common worldwide disease in homeothermic animals, including birds and humans. The aim of the present study was to determine the presence of antibodies against T. gondii in the armadillo Chaetophractus villosus in the province of La Pampa, Argentina. Serum samples were collected from 150 individuals (70 males and 80 females). For serological detection of T. gondii, a latex agglutination test was first performed and then positive sera were confirmed with an indirect hemagglutination test, using 1:4 to 1:64 dilutions. Results showed that 27% (41) of the samples presented titers for antibodies against T. gondii. There were not significant differences between the presence of antibodies against T. gondii and age or sexes of the armadillos. Results show that presence of T. gondii antibodies in armadillos were associated with presence of pigs, and sheep, however there was not association with chickens and dairy cattle in capture site. T. gondii has an important presence in C. villosus population, suggesting a potential zoonotic risk for humans and wildlife animals when C. villosus meats are consumed raw or undercooked. This is the first record of the presence of antibodies against T. gondii in C. villosus.

Cucullanus bagre Petter, 1974 (Cucullanidae) is redescribed based on the examination of newly collected material and the revaluation of type specimens, using light and for the first time scanning electron microscopy. Parasite specimens were collected from the intestine of the coco sea catfish Bagre bagre (Linnaeus) (Ariidae) off the South Atlantic coast, State of Rio de Janeiro, Brazil. Detailed morphological analysis revealed the presence of an unpaired papilla just anterior to the cloacal protrusion in male, that was overlooked in the original description. Descriptions of the adult female and eggs are provided for the first time; these gravid specimens have the general morphometry quite similar to that of juveniles. Additionally, detailed description and illustration of sclerotized plates present on the oesophastome is provided. The far posterior location of the excretory pore with thick-walled excretory duct, the conspicuous protrusions present on both anal and cloacal regions and the number and arrangement of caudal papillae in males are considered to be important diagnostic features for C. bagre. All the specimens from type material were together in one vial and not separated in two vials as stated in the original description, making it impossible to determine which is the holotype, allotype or paratypes. This is the first report of C. bagre in Brazil, which expands its geographical distribution to South Atlantic waters.

Toxascaris leonina (Linstow, 1902) is a most common intestinal parasite of various animals in Felidae and Canidae. In the present paper, light and scanning electron microscopy (SEM) were used to study the morphological aspects of adult worms of this nematode in detail, based on the material collected from Panthera leo (Linnaeus) and Felis lynx (Linnaeus) (Carnivora: Felidae) in China. The results showed that there were some morphometric differences between the present material and the previous studies, including the body size, the width and length of cervical alae, the number of denticles on each lip and the tail length of the female. Previously unreported morphological features were also revealed. These supplementary morphological and morphometric data contributed to a more accurate identification of this worldwide distributed ascarid nematode.

Species of South American subterranean rodents belonging to the genus Ctenomys (commonly called tuco-tucos) are widely distributed across the southern Neotropical region. Despite their relatively well-studied biology and reproductive physiology, current knowledge of their ectoparasite fauna is limited to a few ambiguous studies, based on scattered samples from a small number of host individuals. Ctenomys talarum is the most widely distributed species in the genus. Lice (Phthiraptera) were collected from these tuco-tucos throughout their entire coastal range. Two species, one chewing louse (Gyropus parvus), and one sucking louse (Eulinognathus americanus) were collected. The distribution ranges for both louse species were extended with new locality records. No lice were found in two host populations. Furthermore, co-occurrence of both ectoparasites was not detected.

The study presents the results of a two-year research of protozoan infections of cattles from the Northern-Bačka district (Voјvodina, Serbia). The research was conducted on cow-farms and in private cattle-breeding households, with variuos levels of hygenie. The study included 224 specimens (71 calves, 48 hifers and 105 cows). 22.91% of examined heifers and 15.23% of cows were infected with the ciliate Buxtonella sulcata. The presence of coccidiosis was detected in 45.07% of the calves, 14.58% of the heifers and 6.67% of the cows. The clinical signs of diarrhoea were established in all of the specimens which had more than 1,500 cysts of B.sulcata in 1g of faeces. In the examined calves the number of oocysts was less than 1500 in 1g of faeces, so the coccidiosis did not influence the incidence of the diarrhoea. The results show the first diagnostic record of the ciliate B.sulcata of bovines in Serbia, and this finding contributes to the existing knowledge about the etiopathogenesis of protozoan disease of cattle in this area.

Zuccagnia punctata Cav. (Fabaceae), commonly called jarilla macho or pus-pus, is being used in traditional medicine as an antiseptic, anti-inflammatory and to relieve muscle and bone pain. The aim of this work was to study the anthelmintic effects of three structurally related flavonoids present in aerial parts of Z. punctata Cav. The biological activity of the flavonoids 7-hydroxyflavanone (HF), 3,7-dihydroxyflavone (DHF) and 2´,4´-dihydroxychalcone (DHC) was examined in the free-living nematode Caenorhabditis elegans. Our results showed that among the assayed flavonoids, only DHC showed an anthelmintic effect and alteration of egg hatching and larval development processes in C. elegans. DHC was able to kill 50% of adult nematodes at a concentration of 17 μg/mL. The effect on larval development was observed after 48 h in the presence of 25 and 50 μg/mL DHC, where 33.4 and 73.4% of nematodes remained in the L3 stage or younger. New therapeutic drugs with good efficacy against drug-resistant nematodes are urgently needed. Therefore, DHC, a natural compound present in Z. punctata, is proposed as a potential anthelmintic drug.

In this study, we found unsporulated coccidia oocysts in passerines from the Nevado de Toluca National Park, Mexico. We captured birds and took samples of their droppings during three field visits. We examined a total of 72 fecal samples and found unsporulated coccidia oocysts in 10 samples from five passerine species: Atlapetes pileatus (3), Cardelina ruber (1), Mniotilta varia (1), Oreothlypis celata (2) and Regulus calendula (3). This appears to be the first recorded study of unsporulated coccidia oocysts in passerine species from Mexico