Insights about echinostomiasis by paleomolecular diagnosis

Echinostomiasis is a zoonosis caused by intestinal trematodes and transmitted by the ingestion of mollusks, crustaceans, fish, amphibians, and reptiles, either raw or poorly cooked. Today human infection is endemic in Southeast Asia and the Far East, but has been reported more recently in other regions of the world. Interestingly eggs identified as Echinostoma sp. were found in coprolites from a mummified body human in Brazil, dated 560 ± 40 BP (before present). However, the specific diagnosis based on morphology of the eggs has not been resolved at the species level. As a follow-up to the previous finding, the current study now aims to standardize the methodology for molecular diagnosis and apply it to the coprolite, using current Echinostoma paraensei-positive feces as the reference, and also the same fecal material dried in a stove as an experimental coprolite model. Isolated eggs of E. paraensei and adult worm were included to verify the sensibility and as positive control, respectively. An adult worm of E. luisreyi was used for comparison. PCR using primers in-house for ITS1 region (126 bp) and cox1 (123 bp) of Echinostoma spp. and subsequent nucleotide sequencing were performed. This is the first molecular paleoparasitological diagnosis for echinostomiasis. The methodology was able to amplify specific DNA fragments for the genus Echinostoma sp. in all samples: adult worm, feces, and a single egg of the parasite, in both the experimental coprolite and archaeological sample. Additionally we observed that ancient DNA can also be retrieved without rehydrating the material. The nucleotide sequences from E. paraensei and E. luisreyi are very similar in the fragment analyzed that difficult the differentiation these species, but DNA sequence analysis recovered in the parasite found in the mummy showed more similarity with the species E. paraensei.     

Helminths as heirlooms and souvenirs: a review of new world paleoparasitology

Evidence from studies on ancient human feces, intestinal contents and organs of preserved bodies has established that Enterobius vermicularis, Trichuris trichiura, Diphyllobothrium spp and probably Trichinella spiralis infected humans in the pre-Columbian New World. These species, and perhaps other common human helminths for which there is not yet convincing evidence, probably accompanied transberingeal immigrants and their dogs, and thus can be seen as heirloom parasites. Early humans on the American continents were affected by helminths of native animals such as Paragonimus and Cryptocotyle, and these have also been found in precontact human remains. Michael Kliks considers that the indigenous parasites infecting early Americans may be viewed as zoonotic souvenirs of some 50 millennia of migrations from Alaska to Patagonia. Among the most serious zoonoses would have been infection by cystic hydatid larvae of echinococcid tapeworms, the many enzootic filarial worms, and a variety of larval trematodes and nematodes.     

Status of intestinal parasites infection among primary school children in Kampongcham,Cambodia

A survey was made to find the extent of intestinal parasite infection in Kampongcham, Cambodia in February 2002. A total of 251 fecal specimens were collected from Tonlebat primary school children and examined by formalin-ether sedimentation technique. The overall infection rate of intestinal parasite was 54.2% (males, 57.3%; females, 50.8%). The infection rate of intestinal helminths by the species were as follows: Ascaris lumbricoides 26.3%, Echinostoma sp. 15.6%, hookworm 6.4%, Opisthorchis sp. 4.0%, Rhabditis sp. 2.4%, and Trichuris trichiura 0.4%. The infection rate of intestinal protozoa were as follows: E. coli 7.6%, G. lamblia 3.2%, I. butschlii 3.2%, and E. histolytica 0.8%. More than two different kinds of parasites were found in 16.7% of the stool samples. All the children infected were treated with albendazole, praziquantel and metronidazole according to parasite species. The results showed that intestinal parasites are highly endemic in this area.     

Complex interactions among a nematode parasite (Daubaylia potomaca), a commensalistic annelid (Chaetogaster limnaei limnaei), and trematode parasites in a snail host (Helisoma anceps)

Many biotic interactions can affect the prevalence and intensity of parasite infections in aquatic snails. Historically, these studies have centered on interactions between trematode parasites or between trematodes and other organisms. The present investigation focuses on the nematode parasite Daubaylia potomaca and its interactions with a commensal, Chaetogaster limnaei limnaei , and a variety of trematode species. It was found that the presence of C. l. limnaei indirectly increased the mean intensity of D. potomaca infections, apparently by acting as a restraint for various trematode parasites, particularly the rediae of Echinostoma sp. In turn, Echinostoma sp. rediae adversely affected the mean intensity of D. potomaca by their consumption of both juvenile and adult nematodes present in tissues of the snail. These organisms not only belong to 3 different phyla but occupy distinct trophic levels as well. The complex interactions among these 3 organisms in the snail host provide an excellent example of biotic interactions influencing the infection dynamics of parasites in aquatic snails.     

Human intestinal parasites in the past: new findings and a review

Almost all known human specific parasites have been found in ancient feces. A review of the paleoparasitological helminth and intestinal protozoa findings available in the literature is presented. We also report the new paleoparasitologic findings from the examination performed in samples collected in New and Old World archaeological sites. New finds of ancylostomid, Ascaris lumbricoides, Trichuris trichiura, Enterobius vermicularis, Trichostrongylus spp., Diphyllobothrium latum, Hymenolepis nana and Acantocephalan eggs are reported. According to the findings, it is probable that A. lumbricoides was originally a human parasite. Human ancylostomids, A. lumbricoides and T. trichiura, found in the New World in pre-Columbian times, have not been introduced into the Americas by land via Beringia. These parasites could not supported the cold climate of the region. Nomadic prehistoric humans that have crossed the Bering Land Bridge from Asia to the Americas in the last glaciation, probably during generations, would have lost these parasites, which life cycles need warm temperatures in the soil to be transmitted from host to host. Alternative routes are discussed for human parasite introduction into the Americas.     

Zoonotic and human parasites of inhabitants of Cueva de los Muertos Chiquitos, Rio Zape Valley, Durango, Mexico

We present the first reconstruction of the parasitoses among the people of the Loma San Gabriel culture, as represented by 36 coprolites excavated from the Cueva de los Muertos Chiquitos in Durango, Mexico. The coprolites date to approximately 1,400-yr-ago. Species identified based on eggs recovered include the trematode Echinostoma sp., the tapeworms Hymenolepis sp. and Dipylidium caninum , and the nematodes Ancylostoma duodenale, Enterobius vermicularis, and Trichuris trichiura. After rehydration and screening, 2 methods were used to recover eggs from these samples including spontaneous sedimentation and flotation. Samples were analyzed by 3 different laboratories for independent verification and comparison of methods. Spontaneous sedimentation resulted in the discovery of hymenolepidid eggs that were not found with flotation. Sedimentation was a more-sensitive indicator of prevalence as well. The modified method of flotation permitted estimation of egg concentration and resulted in the detection of a few specimens not found by sedimentation. The results of both methods showed that 19 (of 36) coprolites contained helminth eggs. Our results detected the presence of pathogenic helminths including hookworms and whipworms. The cestodes found do not cause severe pathology in humans. The early dates of hookworm and whipworm, relative to other findings in the southwest United States, indicate that these parasites arrived relatively late in prehistory in Arizona and New Mexico, probably moving into the area with travelers from Mesoamerica.     

Status of intestinal parasite infections among children in Bat Dambang, Cambodia

A survey was conducted to determine the extent of intestinal parasite infection in Bat Dambang, Cambodia in March 2004. A total of 623 fecal specimens was collected from kindergarten and schoolchildren and examined using the formalin-ether sedimentation technique. The overall infection rate of intestinal parasites was 25.7% (boys, 26.2%; girls, 25.1%), and the infection rates of intestinal helminthes by species were as follows: Echinostoma sp. 4.8%, hookworm 3.4%, Hymenolepis nana 1.3%, and Rhabditis sp. 1.3%. The infection rates of intestinal protozoa were; Entamoeba coli 4.8%, Giardia lamblia 2.9%, Iodamoeba butschlii 1.4%, Entamoeba polecki 1.1%, and Entamoeba histolytica 0.8%. There were no egg positive cases of Ascaris lumbricoides or Trichuris trichiura. All children infected were treated with albendazole, praziquantel, or metronidazole according to parasite species. The results showed that intestinal parasites are highly endemic in Bat Dambang, Cambodia.     

Hematological values and parasite fauna in free-rangingMacaca hecki and theM. hecki/M. tonkeana hybrid group of Sulawesi Island, Indonesia

Hematological values and parasitological fauna of free-rangingMacaca hecki and the hybrid group betweenM. hecki/M. tonkeana of Sulawesi Island, Indonesia, were investigated. The hematological values, especially the red cell number (RBC), were lower than those of other macaque species, indicating that Sulawesi macaques are slightly anemic. Several parasites including Plasmodium sp., trombiculid mites, andTrichuris trichiura were identified. Although infection by Plasmodium was observed with considerable frequency, no clear relationship between its infection and the occurrence of anemia was found. Trombiculid mites and eggs ofAnatrichosoma sp. were detected in foci of the ears of most monkeys. The infection with a trombiculid mite is the first recorded occurrence in free-ranging wild Sulawesi macaques. Gastrointestinal parasites were identified from their eggs in fecal samples, where five species of nematoda and one trematoda species were found.     

The finding of eggs of Diphyllobothrium in human coprolites (4,100-1,950 B.C.) from northern Chile

Twenty six coprolites from an archaeological site in the province of Iquique, northern Chile, were examined for parasites. Coprolites were found in two excavation units, I and II (Tiliviche site), dated respectively at 5,900 B.C. to 4,110 B.C. and 4,110 B.C. to 1,950 B.C., and identified as of human origin. Only at the unit II coprolites containing helminth eggs identified as Diphyllobothrium pacificum were found. The presence of this tapeworm, a parasite of the American Sea Lion, in human coprolites, points to a diet which included marine fishes and provides information on the antiquity of infection by Diphyllobothrium pacificum. It is interesting to note that Baer (1969) suggests the presence of this tapeworm in pre-Columbian populations when diagnosing the first human cases in today's population in Peru.     

Infection status with helminthes in feral cats purchased from a market in Busan, Republic of Korea

The present study was performed to investigate the infection status with helminth in a group of feral cats in Korea. More than 29 helminth species including adults or eggs were detected in visceral and fecal samples of the examined cats. Among these were a host of nematodes, including toxocarids, Ancylostoma sp. and the larva of Anisakis simplex; trematodes, including Clonorchis sinensis, Paragonimus westermani, Eurytrema pancreaticum, Pharyngostomum cordatum, Metagonimus spp., Heterophyes nocens, Pygidiopsis summa, Heterophyopsis continua, Stictodora fuscata, Stictodora lari, Acanthotrema felis, Stellantchasmus falcatus, Centrocestus armatus, Procerovum varium, Cryptocotyle sp., Echinostoma revolutum, Echinostoma hortense, Echinochasmus japonicus, Stephanoprora sp., Plagiorchis muris, Neodiplostomum sp. and diplostomulum. We also detected a variety of cestodes, including Spirometra erinacei, Taenia taeniaeformis and unidentified species of tapeworm. We also found examples of the acanthocephalan, Bolbosoma sp. In our assessment of the stools, we detected at least 12 species of helminth eggs. These findings confirmed that feral cats in Korea are infected with a variety of helminth parasite species. Furthermore, among the helminths detected, E. pancreaticum, S. fuscata, S. lari, A. felis, S. falcatus, C. armatus, P. varium, Cryptocotyle sp., E. revolutum, E. japonicus, Stephanoprora sp., P. muris, Neodiplostomum sp. and Bolbosoma sp. represent helminth fauna which have not been reported previously in feral cats in the Republic of Korea.     

Microsporidian Parasites of Trematode Larvae from Aquatic Snails in West Malaysia

In a survey of microsporidian parasites of trematode larvae of Malaysian fresh-water snails, species of Nose-matidae were found 7 times in rediae from 425 Lymnaea rubiginosa, 6 times in rediae from 97 Indoplanorbis exustus and not at all in other snails. There were 3 species of Microsporida in all: one, found in Echinoparyphium dunni, Echinostoma hystricosum and Echinostoma malayanum was identified as Nosema eurytremae Canning; another, which was found in E. dunni and Echinostoma audyi, had paired nuclei (diplokaryon form) in sporogony only and was named a new species, Nosema vasicola sp.n.; the 3rd, from E. hystricosum, lacked diplokaryon nuclei throughout development, was considered to belong to a new genus and was named Unikaryon piriformis gen.n., sp.n. Nosema eurytremae was transmitted experimentally to Fasciola hepatica rediae in Lymnaea truncatula.     

Changes in parasite transmission stage excretion after pheasant release

The production of parasite transmission stages was investigated in the faeces of 77 farm-bred ring-necked pheasants (Phasianus colchicus). Coccidian oocysts (Eimeria sp.), and nematode eggs (Heterakis sp., and Capillaria-like eggs) were recovered before and after release but all birds were treated prior to release. Treatment with fenbendazole significantly reduced the abundance of transmission-stage excretion for all parasites, and reduced the prevalence in the case of Eimeria sp. and Heterakis sp. Nonetheless, a significant increase in the excretion abundance for all parasites and in the prevalence of Eimeria sp. and Heterakis sp. was found after release. Eggs of Ascaridia sp. were found only after releasing, suggesting infection ocurred in the wild. A negative relationship was found between the pheasant body condition and Heterakis excretion abundance and a higher abundance of Capillaria sp. eggs in female birds. No significant relationship was found between parasite excretion abundance and pheasant survival. Despite this, results suggest that an increase in the excretion of parasite transmission stages follows the release of captive pheasants into the wild. This can in part explain restocking failures, but also means that autochtonous free-living birds may become exposed to new and potentially harmful pathogens. To avoid these risks it is proposed that improved prophylactic measures should be taken.     

Parasites with zoonotic potential found in commercially important fish in Tamaulipas,Northeastern Mexico

Human population is exposed to numerous parasitic ichthyozoonoses. Although Tamaulipas state (northeastern Mexico) is well known for its fishing and aquaculture industry, there are few reports of this type of zoonosis. Therefore, it is imperative to investigate whether the parasites that affect these fish may represent a zoonotic risk for the inhabitants of the area.The objective of this study was to identify molecular and/or morphologically muscle parasites of fish from coastal locations in Tamaulipas, Mexico, and assess the risk of infection for humans. Between 2017 and 2018, 764 individual fish belonging to 28 species were examined for parasites. Collected worms were processed for their identification using morphological characteristics. In addition, partial sequences of the large subunit (28S) ribosomal RNA gene were obtained from some species to corroborate their identity. Prevalence and mean intensity of all registered infections were calculated. A total of seven species of parasites were found: cestodes (Poecilancistrium caryophyllum), trematodes (Clinostomum tataxumui, Clinostomum cichlidorum), nematodes (Eustrongylides sp., Contracaecum sp.) and pentastomids (Sebekia purdieae, Sebekia sp.). Parasites infected 10 species belonging to different fish families (Ariidae, Centrarchidae, Centropomidae, Cichlidae, Eleotridae, Ictaluridae, Mugilidae and Sciaenidae). Congeneric species of parasites or related to those registered in this study have been identified as zoonotic agents in other regions of the world. Despite the low levels of infection (2.6–16.6% prevalence and 1–5.5 parasites per infected host), there is a latent risk of transmission to humans, so it is recommended to avoid eating raw or undercooked fish meat.     

生态旅游对川金丝猴肠道寄生虫的影响

非人灵长类接触人类或受人类活动干扰导致肠道寄生虫与宿主关系失衡,这种影响造成肠道寄生虫感染,进而增加肠道寄生虫种类和荷虫量。为检验生态旅游对川金丝猴（Rhinopithecus roxellana）肠道寄生虫的种类和承载是否有影响,本研究收集神农架国家级自然保护区川金丝猴投食群和野生群粪便样品,采用福尔马林乙酸乙酯沉淀法,共发现5种肠道寄生虫：阿米巴原虫（Amoeba）,鞭虫属（Trichuris sp.）,蛔虫（Ascaris lumbricoides）,蛲虫（Enterobius vermicularis）,钩虫（Hookworms）。结果显示,投食群的阿米巴和蛲虫的荷虫量高于野生群;蛔虫在投食群和野生群均有发现,但两者之间无显著性差异;鞭虫和钩虫只在投食群中发现。投食群部分肠道寄生虫具有高荷虫量和新增的两个种类是由于非人灵长类直接或间接与人类接触（生态旅游等）所导致的。     

Cryptosporidium sp. skunk genotype in wild raccoons (Procyon lotor) naturally infected with Baylisascaris procyonis from Central Germany

Cryptosporidium spp. are apicomplexan parasites of public health concern. They are one of the main causes of intestinal diseases in humans and animals. Contaminated water is among the main sources of infection for humans and mammals. Raccoons are an introduced species in Germany. They are anthropogenic adapters with a natural affinity for water bodies. We collected samples from wild raccoons in the Federal States of Saxony and Thuringia, Central Germany. Through molecular genotyping, we found Cryptosporidium sp. skunk genotype in one raccoon from Saxony (1/24) and in one animal from Thuringia (1/27). Both raccoons were also infected with the zoonotic nematode Baylisascaris procyonis. This is the first report of co-infection with these two parasites in raccoons from Germany. Our study highlights the potential of these animals as carriers of zoonotic pathogens. Since raccoons can thrive in human settlements, this study provides data that can be used as a baseline for preventive programs.     

Paleoparasitological remains revealed by seven historic contexts from "Place d'Armes", Namur, Belgium

Human occupation for several centuries was recorded in the archaeological layers of "Place d'Armes", Namur, Belgium. Preventive archaeological excavations were carried out between 1996/1997 and seven historical strata were observed, from Gallo-Roman period up to Modern Times. Soil samples from cesspools, latrines, and structures-like were studied and revealed intestinal parasite eggs in the different archaeological contexts. Ascaris lumbricoides, A. suum, Trichuris trichiura, T. suis. Taenia sp., Fasciola hepatica, Diphyllobothrium sp., Capillaria sp. and Oxyuris equi eggs were found. Paleoparasitology confirmed the use of structures as latrines or cesspit as firstly supposed by the archaeologists. Medieval latrines were not only used for rejection of human excrements. The finding of Ascaris sp. and Trichuris sp. eggs may point to human's or wild swine's feces. Gallo-Roman people used to eat wild boar. Therefore, both A. suum and T. suis, or A. lumbricoides and T. trichuris, may be present, considering a swine carcass recovered into a cesspit. Careful sediment analysis may reveal its origin, although parasites of domestic animals can be found together with those of human's. Taenia sp. eggs identified in latrine samples indicate ingestion of uncooked beef with cysticercoid larvae. F. hepatica eggs suggest the ingestion of raw contaminated vegetables and Diphyllobothrium sp. eggs indicate contaminated fresh-water fish consumption. Ascaris sp. and Trichuris sp. eggs indicate fecal-oral infection by human and/or animal excrements.     

Crab louse infestation in pre-Columbian America

Until now, Pthirus pubis infestation in ancient human populations had only been recorded in the Old World. We found crab lice on South American mummified bodies from the Atacama Desert region. Crab louse eggs were found attached to the pubic hairs of a 2,000-yr-old Chilean mummy. Well-preserved adults were found in sediment and clothing from a Peruvian mummy dated 1,000 yr ago. Paleoparasitological evidence expands the knowledge of the distribution of this ectoparasite in ancient populations. As with many other parasites, pubic lice recorded in Andean populations show the antiquity of this parasite in the New World. It is likely that P. pubis entered the continent with early human migration to the New World.     

Molecular paleoparasitological diagnosis of Ascaris sp. from coprolites: new scenery of ascariasis in pre-Colombian South America times

Paleoparasitological studies using microscopy showed that Ascarisand Trichuris trichiura are the human intestinal parasites most found in archaeological sites. However, in pre-Columbian South American archaeological sites, Ascaris is rare. In this work we standardized a molecular methodology for Ascaris diagnosis directly from ancient DNA retrieved from coprolites. Using cythochrome b gene (142 bp) target, ancient DNA sequences were retrieved from South American samples, negative by microscopy. Moreover, the methodology applied was sensitive enough to detect ancient DNA extracted from 30 Ascaris eggs from an European coprolite. These results revealed a new scenery for the paleodistribution of Ascaris in South America.     

Nematodirus nemorhaedi sp. n. (Nematoda: Trichostrongylidae) from the Amur goral Nemorhaedus caudatus raddeanus]

Nematodirus nemorhaedi sp. n. is described. The new species was found in the small intestine in three long-tailed gorals from the Maritime Territory. The species differs from other species of the genus, parasites of ungulates, by very large (over 1.5 mm) spicules and characteristic structure of their distal end in males; by long (about 1 mm) ovijector in females and by a specific combination of characters of synloph, genital bursa in males, sizes of body, structure of vulva, sizes and shape of eggs and tail in females.     

Immunodetection of Helicobacter sp. and the associated expression of ABO blood group antigens in the gastric mucosa of captive and free-living New World primates in the Amazon region

The histo-blood group ABH antigens were first described in humans. These antigens are only present on erythrocytes from great apes and humans, while in more primitive animals they are found in tissues and body fluids. The ABH antigens are mainly distributed in tissues exposed to the external environment and potentially serve as ligands for pathogens or inhibitors of tissue connections. The objective of this paper was two-fold: (i) to determine the presence of Helicobacter sp. in the gastric mucosa of 16 captive and 24 free-living New World monkeys and (ii) to evaluate the presence of histopathological alterations related to bacterial infection and the associated expression of ABH antigens in the tissue. Stomach tissues from 13 species of monkey were assessed using haematoxylin-eosin and modified Gram staining (Hucker) methods. An immunohistochemical analysis of the tissue revealed the presence of infectious bacteria that were characteristic of the genus Helicobacter sp. The results demonstrate that various species of monkey might be naturally infected with the Helicobacter sp. and that there is an increased susceptibility to infection. This study serves as a comparative analysis of infection between human and non-human primates and indicates the presence of a new species of Helicobacter.