Host feces, olfactory beacon guiding aggregation of intestinal parasites Gasterophilus pecorum (Diptera: Gasterophilidae).

The aim of this study was to identify the aggregation sites and transmission characteristics of Gasterophilus pecorum, the dominant pathogen of endangered equines in desert steppe. Therefore, we tested with a four-arm olfactometer the olfactory response of the G. pecorum adults to the odors that have a great impact on their life cycle, and also investigated the occurrence sites of the adults in the area where the Przewalski's horse (Equus przewalskii) roam frequently during the peak period of G. pecorum infection. The results of four-directional olfactory test showed that the fresh horse feces had a stronger attraction rate on both male (50.4%) and female flies (38.2%). Stipa caucasica, the only oviposition plant where G. pecorum lay eggs, had a better attraction effect on females than that on males. And the attraction rates of S. caucasica to G. pecorum females in the early growth stage (Stipa I) and mid-growth stage (Stipa II) were 32.8% and 36.8%, respectively. In addition, the two-directional olfactory test showed that the attraction rate of males to fresh horse feces (68.90%) was higher than that to Stipa II (31.10%), and females also showed similar olfactory responses. Moreover, in our field investigation, 68.29% of G. pecorum adults were collected from around the horse feces. The results of laboratory test and field investigation implied that the location mechanism of G. pecorum aggregation for mating is related to the orientation of horse feces. The horse feces and the vicinity are the key contamination areas of G. pecorum, and it is also the areas where horses are seriously infected with G. pecorum. Those fresh feces, which gather abundant information about the host, naturally had the greatest chance of contacting with the host; G. pecorum adults create the opportunity to enter directly into the host's mouth and infect the host by laying eggs on S. caucasica, which is the most favorite plant of the host in this area. These characteristics are one of the main reasons why G. pecorum has become the dominant species under the condition of sparse vegetation in desert steppe.

Parasitic communities of Hoplosternum littorale (Hancock, 1828) as indicators of environmental impact.

The parasites are indicative of several biological aspects of their hosts, including their diet, migration, recruitment and phylogeny, but they can also be direct indicators of the environmental quality. Hoplosternum littorale is a fish that has a high commercial value in some South American countries. The fishes were collected from January to July 2012, in two locations of the Peixe River in the municipality of Anhembi, State of São Paulo, Brazil, which had different environmental and limnological characteristics (river mouth and Lagoa Cabeça de Boi). An amount of 30 fish specimens were captured in each location for the analysis of their parasitic fauna. Nine species of metazoan parasites were collected. Both Brillouin`s diversity index and Margalef`s richness index were greater in the Lagoa Cabeça de Boi. The ratio of heteroxenous and monoxenous parasite species richness (SH/SM) was higher in the Lagoa Cabeça de Boi. The results of the physicochemical analysis of the water showed that the mouth of the Peixe River can be considered an area that suffers more from anthropogenic actions than the Lagoa da Cabeça de Boi. Proteocephalus sp., Gyrodactylus sp., Scleroductus sp. and Capillaria (Neocapillaria) pterophylli are being registered for the first time in this host.

[Metazoan parasites of Micropterus salmoides (Centrarchidae: Perciformes) from reservoirs of Nuevo León, México and their association with condition factor and gender]. / Metazoarios parásitos de Micropterus salmoides (Perciformes: Centrarchidae) de reservorios de Nuevo León, México y su asociación con el factor de condición y sexo.

The largemouth bass Micropterus salmoides is a very valuable fish species for aquaculture and sport fishing; however, there are no systematic studies on fish metazoan parasites in Mexico. The main objective of the present study was to describe the prevalence, abundance, and intensity of M. salmoides metazoan parasites, and their association with fish condition factor and gender. The sample size was composed by 672 hosts, collected between 2011-2013 from the following reservoirs of Nuevo Leon, México: Rodrigo Gómez dam ("La Boca", LB), Cuchillo-Solidaridad dam (CS), Salinillas lagoon (LS), Mariano Escobedo dam ("Sombreretillo", S), and José López Portillo dam ("Cerro Prieto", CP). Living fish were transported to the laboratory; sizes and weights were then recorded to calculate the Fulton condition factor (k). If possible, gender was also recorded. Parasites were detected under stereoscopy, recollected and preserved by traditional techniques. Statistical analysis of association between parasitic load, gender, and Fulton condition factor were calculated, using the X2 and the Student-t tests. Results showed that 12 different metazoans were identified, two flukes (Posthodiplostomum minimum centrarchi and Phyllodistomum pearsei), one tapeworm (Proteocephalus ambloplitis), three roundworms (Contracaecum sp., Spinitectus carolini and Philometra nodulosa), two acantocephalan (Neoechinorhynchus cylindratus and Arhythmorhynchus sp.), one leech (Myzobdella moorei), and three copepods (Ergasilus versicolor; Ergasilus arthrosis and Ergasilus cerastes). HSD Tukey test showed that infected fish from LB were significantly different than LS, CS, CP, and S (P<0.05). Parasites most commonly collected in all five locations were P. m. centrarchi, Contracaecum sp. and E. versicolor. The frequency of P. m. centrarchi was highly significant (P<0.001) than other parasites; furthermore, this parasite showed the highest prevalence (97.5 %), abundance (10.12-83.6), and intensity (15.44-88.5). Statistical dependence was found between intensity and abundance of metazoans with regard to female gender and size on five reservoirs (t=143.82, P=0.0001; t=2.6, P=0.01, respectively). In addition, the abundance and intensity showed statistical dependence with regard to k, which was proportionally significant. New locality records for Nuevo Leon include P. m. centrarchi, P. pearsei, P. ambloplitis, Contracaecum sp., Arhythmorhynchus sp., and E. cerastes. Furthermore, P. nodulosa, S. carolini, N. cylindratus, and E. arthrosis were new records for Mexico.

Parasites of wild rabbits (Oryctolagus cuniculus) from an urban area in Germany, in relation to worldwide results.

The European rabbit (Oryctolagus cuniculus) belongs to the most invasive and successful mammalian species, which is distributed nearly worldwide. In Europe, they inhabit broad parts of the mainland and subsequently reached several European islands via anthropogenic diversion. Rabbits can also serve as hosts for numerous parasite species. The parasite and pathogen fauna of O. cuniculus have been well documented in various European countries, although studies in Germany are scarce. Until now, a comprehensive survey combining recent international studies over parasite fauna of wild rabbits had not been conducted. We examined 50 wild rabbits from an urban area near Aachen (Germany) to identify their metazoan parasite fauna, and then compared our findings to previous international investigations. A total of nine parasite species were isolated consisting of four endoparasite species (Cittotaenia denticulata, Graphidium strigosum, Passalurus ambiguus, and Trichostrongylus retortaeformis) and five ectoparasite species (Cheyletiella parasitivorax, Ixodes ricinus, Leporacarus gibbus, Haemodipsus ventricosus, and Spilopsyllus cuniculi). Among the ectoparasites were two verifiable human pathogenic species and two potentially pathogenic species. In comparison to previous studies, a high number of similarities in composition of helminth species fauna were revealed. Furthermore, our results showed partial agreement with international surveys in prevalence and mean intensity of the parasites C. denticulata, G. strigosum, P. ambiguus, and T. retortaeformis.

The parasite fauna of the Patagonian toothfish Dissostichus eleginoides off the Falkland Islands.

The parasite fauna of juvenile Dissostichus eleginoides, while they inhabit the Falkland's shelf, was examined, giving new detailed information on spatial, ontogenic and seasonal variations. A total of 24,943 parasites from 15 different taxa were found in the stomach of 502 individual fish. Parasite species composition and abundance allowed separation of toothfish by area between the north-west and south-east of the Falklands. The digenean, Elytrophalloides oatesi, and the nematodes, Hysterothylacium spp. and Anisakis spp., were the most common, all with a prevalence >20%. For some seasons ontogenic changes in abundance were significant in these three parasite taxa, and this is discussed in terms of ontogenic and seasonal changes in diet. Elytrophalloides oatesi and Hysterothylacium spp. showed spatial and seasonal differences in abundance with greater numbers in the warmer waters of the north-west and during the summer months. Differences in abundance of E. oatesi between the Falklands and other regions indicate its potential for use as a biological tag to study toothfish movements and population structure; however, more seasonal data would be required before this technique could be used.

Microplastics in grey seal (Halichoerus grypus) intestines: Are they associated with parasite aggregations?

Between 2012 and 2015, 13 grey seals were recovered from trammel nets targeting monkfish and rays off the south coast of Ireland. Incidence and distribution of microplastics were investigated along the intestines of bycaught seals. No macrodebris items were found, whereas microplastics were detected in all seals. A total of 363 microplastics items were identified (85% fibers, 14% fragments, 1% films). Estimation of microplastic ingestion based on prey ingestion (245 particles) was lower than the observed data. Acantocephala parasites (n = 1543) were found in 12 seals, with an average of 74.5 ±â€¯67.7 parasites per seal. Distribution of microplastics varied between seals, although microplastics tended to accumulate in areas where more parasites were aggregated; however, there was no significant relationship between the number of parasites and microplastics was found. Seals recovered from nets appear to be a good source to monitor the incidence of microplastic pollution within the coastal food webs.

Ultrafiltration-based techniques for rapid and simultaneous concentration of multiple microbe classes from 100-L tap water samples.

This study focused on ultrafiltration as a technique for simultaneously concentrating and recovering viruses, bacteria and parasites in 100-L drinking water samples. A chemical dispersant, sodium polyphosphate, and Tween 80 were used to increase microbial recovery efficiencies. Secondary concentration was performed to reduce sample volumes to 3-5 mL for analysis using tissue culture, microscopy, and real-time PCR and RT-PCR. At seeding levels of 100-1000 (CFU, PFU, oocysts, or particles), a "high-flux" ultrafiltration procedure was found to achieve mean recoveries of 51-94% of simultaneously seeded MS2 bacteriophage, echovirus 1, Salmonella enterica subsp. enterica serovar Typhimurium, Bacillus atrophaeus subsp. globigii endospores, Cryptosporidium parvum oocysts, and 4.5-mum microspheres. When 4-7% of the final sample concentrate volume was assayed using real-time PCR and RT-PCR, overall method sensitivities were <100 C. parvum oocysts, <240 PFU echovirus 1, <100 CFU Salmonella and approximately 160 CFU B. atrophaeus spores in 100-L drinking water samples. The "high-flux" ultrafiltration procedure required approximately 2 h, including time required for backflushing. Secondary concentration procedures required an additional 1-3 h, while nucleic acid extraction and real-time PCR procedures required an additional 2-2.5 h. Thus, this study demonstrated that efficient recovery and sensitive detection of diverse microbes in 100-L drinking water samples could be achieved within 5-8 h using ultrafiltration, rapid secondary processing techniques, and real-time PCR.

Metazoan parasite communities of two deep-sea elasmobranchs: the southern lanternshark, Etmopterus granulosus, and the largenose catshark, Apristurus nasutus, in the Southeastern Pacific Ocean.

Two deep-sea shark species were obtained as by-catch of the local fishery of the Patagonian toothfish, Dissostichus eleginoides, at depths ranging from 1000 to 2200 m off central and northern Chile. A total of 19 parasite taxa were found in 133 specimens of the southern lanternshark, Etmopterus granulosus, (n = 120) and largenose catshark, Apristurus nasutus, (n = 13). Fourteen taxa (four Monogenea, one Digenea, four Cestoda, one Nematoda, two Copepoda, one Annelida and one Thecostraca) were found in E. granulosus, whereas five taxa (one Monogenea, two Cestoda and two Nematoda) were found in A. nasutus. Representatives of Cestoda showed higher values of prevalence and a greater intensity of infection; this pattern is consistent with reports for elasmobranchs, but the monogenean richness was higher than that previously reported for related deep-sea sharks. Regarding E. granulosus, a positive and significant correlation between host length and abundance was found for six (four ectoparasites, one mesoparasite, and one endoparasite) of the 14 taxa recorded, but prevalence was significantly correlated with host length only for the monogenean Asthenocotyle sp. Although the sample size for A. nasutus was limited, we compared richness, abundance, diversity and evenness at the infracommunity and component community levels. All of these variables were higher for E. granulosus, but at the infracommunity level, abundance was higher for A. nasutus. All the parasite taxa (except two) represent new host and geographical records.

Variations in the parasite fauna of the Patagonian toothfish (Dissostichus eleginoides Smitt, 1898), with length, season, and depth of habitat around the Falkland Islands.

The parasite fauna of Dissostichus eleginoides was examined from locations around the Falkland Islands. In total, of 11,362 individual parasites of 27 taxa were recovered from 105 fish. Two species, Ceratomyxa dissostichi and Sphaerospora dissostichi, represent new host records. The nematode Ascarophis nototheniae and the larval acanothocephalan Corynosoma bullosum were found to be new locality records and add to the knowledge of the biogeography and host specificity of parasites on the Patagonian Shelf. There were no significant differences in the mean abundance and prevalence of parasites recovered between sexes. Therefore, sex was not considered in further analysis and the data were pooled. Cysts of unknown etiology (CUE), the monogenean Neopavlovskioides georgianus, the larval acanthocephalan Corynosoma bullosum, and the digenean Neolepidapedon magnatestis had significant positive correlations with increasing host length. The larval Trypanorhynch cestode Grillotia erinaceus and the digenean Elytrophalloides oatesi showed significant negative correlations with increasing host length. CUEs, N. georgianus, the digenean Gonocerca physidis and E. oatesi showed statistically significant prevalence between summer, winter, and spring. The effect of depth on parasite communities was also examined, initially using a linear discriminant function analysis. The prevalence of individual parasites was then compared between depth strata using the chi-square test. The parasite communities on the shelf and deep water (> 1,000 m) were found to be different, whereas those caught at intermediate depths on the shelf slope were found to have parasite communities that were intermediate, containing a mixture of shelf and deeper-water parasites. The causes of the variations in parasite faunas in association with these intrinsic and extrinsic factors are discussed.

Electrochemical magnetic microbeads-based biosensor for point-of-care serodiagnosis of infectious diseases.

Access to appropriate diagnostic tools is an essential component in the evaluation and improvement of global health. Additionally, timely detection of infectious agents is critical in early diagnosis and treatment of infectious diseases. Conventional pathogen detection methods such as culturing, enzyme linked immunosorbent assay (ELISA) or polymerase chain reaction (PCR) require long assay times, and complex and expensive instruments making them not adaptable to point-of-care (PoC) needs at resource-constrained places and primary care settings. Therefore, there is an unmet need to develop portable, simple, rapid, and accurate methods for PoC detection of infections. Here, we present the development and validation of a portable, robust and inexpensive electrochemical magnetic microbeads-based biosensor (EMBIA) platform for PoC serodiagnosis of infectious diseases caused by different types of microorganisms (parasitic protozoa, bacteria and viruses). We demonstrate the potential use of the EMBIA platform for in situ diagnosis of human (Chagas disease and human brucellosis) and animal (bovine brucellosis and foot-and-mouth disease) infections clearly differentiating infected from non-infected individuals or animals. For Chagas disease, a more extensive validation of the test was performed showing that the EMBIA platform displayed an excellent diagnostic performance almost indistinguishable, in terms of specificity and sensitivity, from a fluorescent immunomagnetic assay and the conventional ELISA using the same combination of antigens. This platform technology could potentially be applicable to diagnose other infectious and non-infectious diseases as well as detection and/or quantification of biomarkers at the POC and primary care settings.

Cell-Free DNA as a Diagnostic Tool for Human Parasitic Infections.

Parasites often cause devastating diseases and represent a significant public health and economic burden. More accurate and convenient diagnostic tools are needed in support of parasite control programmes in endemic regions, and for rapid point-of-care diagnosis in nonendemic areas. The detection of cell-free DNA (cfDNA) is a relatively new concept that is being applied in the current armamentarium of diagnostics. Here, we review the application of cfDNA detection with nucleic acid amplification tests for the diagnosis and evaluation of different human parasitic infections and highlight the significant benefits of the approach using non-invasive clinical samples.

Parasitic communities of Hoplosternum littorale (Hancock, 1828) as indicators of environmental impact

ABSTRACT The parasites are indicative of several biological aspects of their hosts, including their diet, migration, recruitment and phylogeny, but they can also be direct indicators of the environmental quality. Hoplosternum littorale is a fish that has a high commercial value in some South American countries. The fishes were collected from January to July 2012, in two locations of the Peixe River in the municipality of Anhembi, State of São Paulo, Brazil, which had different environmental and limnological characteristics (river mouth and Lagoa Cabeça de Boi). An amount of 30 fish specimens were captured in each location for the analysis of their parasitic fauna. Nine species of metazoan parasites were collected. Both Brillouin`s diversity index and Margalef`s richness index were greater in the Lagoa Cabeça de Boi. The ratio of heteroxenous and monoxenous parasite species richness (SH/SM) was higher in the Lagoa Cabeça de Boi. The results of the physicochemical analysis of the water showed that the mouth of the Peixe River can be considered an area that suffers more from anthropogenic actions than the Lagoa da Cabeça de Boi. Proteocephalus sp., Gyrodactylus sp., Scleroductus sp. and Capillaria (Neocapillaria) pterophylli are being registered for the first time in this host.

Metazoarios parásitos de Micropterus salmoides (Perciformes: Centrarchidae) de reservorios de Nuevo León, México y su asociación con el factor de condición y sexo / Metazoan parasites of Micropterus salmoides (Centrarchidae: Perciformes) from reservoirs of Nuevo León, México and their association with condition factor and gender

ResumenMicropterus salmoides es una especie muy apreciada en la acuacultura y pesca deportiva, sin embargo, no existen estudios sistemáticos sobre los parásitos en México. El objetivo principal del presente estudio fue describir la prevalencia, abundancia e intensidad de los metazoarios parásitos de M. salmoides y su asociación con el factor de condición y el sexo. El tamaño de muestra fue de 672 hospederos, recolectados entre 2011-2013 de los siguientes reservorios de Nuevo León, México: Presa Rodrigo Gómez (“La Boca”, LB); Presa Cuchillo-Solidaridad (CS), Laguna Salinillas (LS), Presa Mariano Escobedo (“Sombreretillo”, S), y Presa José López Portillo (“Cerro Prieto”, CP). Los peces fueron transportados vivos al laboratorio, se les registró el tamaño y peso para calcular el coeficiente de condición de Fulton (k). El sexo también se registró cuando fue posible. Los parásitos fueron detectados con el estereoscopio, recolectados y preservados con las técnicas parasitológicas tradicionales. Se calcularon los análisis estadísticos de asociación entre la carga parasitaria, sexo y factor de condición usando X2 y la prueba de t de Student. Los resultados mostraron la identificación de 12 metazoarios diferentes: dos tremátodos (Posthodiplostomum minimum centrarchi y Phyllodistomum pearsei); un céstodo (Proteocephalus ambloplitis); tres nemátodos (Contracaecum sp.; Spinitectus carolini y Philometra nodulosa); dos acantocéfalos (Neoechinorhynchus cylindratus y Arhythmorhynchus sp.); un hirudineo (Myzobdella moorei) y tres copépodos (Ergasilus versicolor; Ergasilus arthrosis y Ergasilus cerastes). La prueba de Tukey HSD mostró que los peces infectados de LB fueron significativamente diferentes a los de LS, CS, CP y S (P < 0.05). Los parásitos que se recolectaron más comúnmente en las cinco localidades fueron: P. m. centrarchi, Contracaecum sp., y E. versicolor. La frecuencia de P. m. centrarchi fue altamente significativa (P < 0.001) en comparación con los otros parásitos; además presentó la más elevada prevalencia (97.5 %), abundancia (10.12-83.6) e intensidad (15.4488.5). Se encontró asociación estadística entre la intensidad y abundancia de los metazoarios con respecto al sexo femenino y tamaño en los cinco reservorios (t= 143.82, P = 0.0001; t= 2.6, P= 0.01, respectivamente). Además, la abundancia e intensidad mostraron asociación estadística con respecto a k, lo cual fue proporcionalmente significativo. Se incluyen nuevos registros de localidad para P. m. centrarchi, P. pearsei, P. ambloplitis, Contracaecum sp., Arhythmorhynchus sp. y E. cerastes. Además, P. nodulosa, S. carolini, N. cylindratus, y E. arthrosis se consideran como nuevos registros para México.

AbstractThe largemouth bass Micropterus salmoides is a very valuable fish species for aquaculture and sport fishing; however, there are no systematic studies on fish metazoan parasites in Mexico. The main objective of the present study was to describe the prevalence, abundance, and intensity of M. salmoides metazoan parasites, and their association with fish condition factor and gender. The sample size was composed by 672 hosts, collected between 2011-2013 from the following reservoirs of Nuevo Leon, México: Rodrigo Gómez dam (“La Boca”, LB), Cuchillo-Solidaridad dam (CS), Salinillas lagoon (LS), Mariano Escobedo dam (“Sombreretillo”, S), and José López Portillo dam (“Cerro Prieto”, CP). Living fish were transported to the laboratory; sizes and weights were then recorded to calculate the Fulton condition factor (k). If possible, gender was also recorded. Parasites were detected under stereoscopy, recollected and preserved by traditional techniques. Statistical analysis of association between parasitic load, gender, and Fulton condition factor were calculated, using the X2 and the Student-t tests. Results showed that 12 different metazoans were identified, two flukes (Posthodiplostomum minimum centrarchi and Phyllodistomum pearsei), one tapeworm (Proteocephalus ambloplitis), three roundworms (Contracaecum sp., Spinitectus carolini and Philometra nodulosa), two acantocephalan (Neoechinorhynchus cylindratus and Arhythmorhynchus sp.), one leech (Myzobdella moorei), and three copepods (Ergasilus versicolor; Ergasilus arthrosis and Ergasilus cerastes). HSD Tukey test showed that infected fish from LB were significantly different than LS, CS, CP, and S (P<0.05). Parasites most commonly collected in all five locations were P. m. centrarchi, Contracaecum sp. and E. versicolor. The frequency of P. m. centrarchi was highly significant (P<0.001) than other parasites; furthermore, this parasite showed the highest prevalence (97.5 %), abundance (10.12-83.6), and intensity (15.44-88.5). Statistical dependence was found between intensity and abundance of metazoans with regard to female gender and size on five reservoirs (t=143.82, P=0.0001; t=2.6, P=0.01, respectively). In addition, the abundance and intensity showed statistical dependence with regard to k, which was proportionally significant. New locality records for Nuevo Leon include P. m. centrarchi, P. pearsei, P. ambloplitis,Contracaecum sp., Arhythmorhynchus sp., and E. cerastes. Furthermore, P. nodulosa, S. carolini, N. cylindratus, and E. arthrosis were new records for Mexico. Rev. Biol. Trop. 64 (2): 559-569. Epub 2016 June 01.

Evaluation of Streck tissue fixative, a nonformalin fixative for preservation of stool samples and subsequent parasitologic examination.

We undertook a study to evaluate Streck tissue fixative (STF) as a substitute for formalin and polyvinyl alcohol (PVA) in fecal preservation. A comparison of formalin, PVA, (mercuric chloride based), and STF was done by aliquoting fecal samples into each fixative. Stool specimens were collected in Haiti, and parasites included Cyclospora cayetanensis, Giardia intestinalis, Entamoeba coli, Iodamoeba butschlii, Endolimax nana, Ascaris lumbricoides, Trichuris trichiura, Strongyloides stercoralis, and Necator americanus. Preserved stools were examined at various predetermined times (1 week, 1 month, and 3 months) to establish the quality of the initial preservation as well as the suitability of the fixative for long-term storage. At each time point, stool samples in fixatives were examined microscopically as follows: (i) in wet mounts (with bright-field and epifluorescence microscopy), (ii) in modified acid-fast-, trichrome-, and safranin-stained smears, and (iii) with two commercial test kits. At the time points examined, morphologic features remained comparable for samples fixed with 10% formalin and STF. For comparisons of STF- and 10% formalin-fixed samples, specific findings showed that Cyclospora oocysts retained full fluorescence, modified acid-fast- and safranin-stained smears of Cryptosporidium and Cyclospora oocysts were equal in staining quality, and results were comparable in the immunofluorescence assay and enzyme immunoassay commercial kits. Stool fixed in STF and stained with trichrome showed less-than-acceptable staining quality compared with stool fixed in PVA. STF provides an excellent substitute for formalin as a fixative in routine examination of stool samples for parasites. However, modifications to the trichrome staining procedures will be necessary to improve the staining quality for protozoal cysts fixed in STF to a level comparable to that with PVA.