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Carolina Davies, Analía Simonazzi, Juan Francisco Micheloud, Paula Gabriela Ragone, Alicia Graciela Cid, Olga Sánchez Negrette, José María Bermúdez, Luis Antonio Parada
Benznidazole and nifurtimox are the only drugs specifically approved for the treatment of Chagas disease. Both compounds are given orally in tablets, but occasionally are ineffective and cause adverse effects. Benznidazole, the first-line treatment in many countries, is a compound with low solubility in water that is administered at high doses for long periods of time. To improve its solubility, we developed a new liquid formulation on the basis of solid dispersions (SD) using the amphiphilic polymer poloxamer 407. Herein we present data on its trypanocidal performance in mouse models of acute and chronic Trypanosoma cruzi infection. SD at doses of 60 or 15 mg/kg per day given with different administration schedules were compared with the commercial formulation (CF; 50 mg/kg per day) and vehicle. The SD performance was assessed by direct parasitemia, total anti-T. cruzi antibodies, and parasitic burden in tissues after 4 or 6 mo posttreatment. The efficacy of the SD was equivalent to the CF but without manifest side effects and hepatotoxicity. Considering our previous data on solubility, together with these on efficacy, this new liquid formulation represents a promising alternative for the treatment of Chagas disease, particularly in cases when dosing poses a challenge, as in infants.
We investigated the mean abundance of helminths and analyzed helminth composition and structure at the infracommunity and component community levels for 3 anuran species (Pleurodema diplolister, Rhinella jimi, and Rhinella granulosa) from the Caatingas, a semiarid Brazilian region characterized by accentuated seasonality and unpredictability of rains. Data were collected during the reproductive period and during drought, when P. diplolister estivated buried underground but R. jimi and R. granulosa remained foraging actively. We expected higher parasitological parameters during the reproductive period when compared to drought for these 3 anurans. We also expected higher parasite infection in the Rhinella species and higher similarity between their helminth parasite communities when compared to the estivating species, P. diplolister. Contrary to our hypothesis, the season was not related to parasite community structure. As predicted, the Rhinella toads shared more similar species composition of parasite communities. These similarities in the composition of the parasite community between Rhinella species could be due to similar temporal/spatial patterns of activity and phylogenetic proximity. Pleurodema diplolister hosted a more restricted helminth fauna, a result that might be associated with estivation restricting the temporal window available to acquire parasites. This study also presents new helminth fauna records for R. jimi and P. diplolister, and the first helminth fauna record of R. granulosa from the Caatingas in semiarid Brazil.
Rodents are reservoirs and hosts of several pathogens around the world, including zoonotic parasite species. This study aimed to determine the occurrence of zoonotic gastrointestinal helminths in rodents captured inside households in a rural community from southern Guatemala. Sixty-nine rodents were captured in 33% (49/148) of the surveyed households, including Rattus rattus, Rattus norvegicus, Mus musculus, and Sigmodon hispidus. Thirty-six percent (25/69) of these rodents (3 Rattus and 22 Mus musculus), from 45% (22/49) of the households, were parasitized with at least 1 gastrointestinal helminth species. Helminths from 6 species were identified: Hymenolepis diminuta, Moniliformis moniliformis, Heterakis spumosa, Nippostrongylus sp., Strongyloides sp., and Syphacia sp. Two zoonotic species were found in Rattus, H. diminuta in R. rattus (1/6), and M. moniliformis in R. norvegicus (1/1). Coinfection with other non-zoonotic helminth parasites, such as He. spumosa and Strongyloides sp., also was observed in the Rattus genus. Mus musculus had only non-zoonotic helminths: He. spumosa, Nippostrongylus sp., and Syphacia sp. being the most common, and He. spumosa (96%) followed by Nippostrongylus sp. (48%), with a higher presence in males than females, with a similar proportion in adult and young individuals. This is the first report of zoonotic and non-zoonotic helminths parasites in rodents from Guatemala.
Waterborne transmission of Toxoplasma gondii is assumed to be enhanced in areas with human-altered landscapes (e.g., urbanization, agriculture) and increased populations of non-native domestic and feral cats (Felis catus). However, little is known concerning T. gondii exposure risks in more natural watersheds (e.g., reduced human footprint, no domestic or feral cats) to establish a baseline for comparisons. In this study, muskrats (Ondatra zibethicus) were used as sentinels to assess baseline T. gondii exposure in a relatively pristine watershed in the Greater Voyageurs Ecosystem, northern Minnesota, during the summers of 2018–2019. Toxoplasma gondii antibodies were assayed in sera of live-trapped muskrats (n = 70) using a modified agglutination test. None of our samples were positive for T. gondii antibodies (P = 0.00, 95% Wald Score Confidence Interval = 0.00–0.05). This study establishes a baseline to compare T. gondii waterborne transmission risks in other human-modified watersheds.
Thelohanellus magnacysta n. sp. (Bivalvulida: Myxobolidae) infects the skeletal muscle of blacktail shiner, Cyprinella venusta Girard, 1856 (Cypriniformes: Cyprinidae) in Bull Creek, Chattahoochee River Basin, eastern Georgia. Although numerous members of ThelohanellusKudo, 1933 have overlapping myxospore dimensions with the new species, it differs from all nominal congeners by polar filament coil number and polar capsule width as well as by lacking a mucous envelope, iodinophilic vacuole, and sutural markings. With the use of novel primers for Myxozoa, a phylogenetic analysis of the small subunit ribosomal DNA (SSU rDNA) suggests that the new species shares a recent common ancestor with a clade of cyprinid-infecting species of Myxobolus Bütschli, 1882 (Bivalvulida: Myxobolidae) and Thelohanellus. Consistent with other published research concerning the systematics of Thelohanellus, this result suggested that Thelohanellus and Myxobolus are polyphyletic and need revision. Histological sections of infected blacktail shiners confirmed that myxospores were only found within a plasmodium and only infected skeletal muscle and that plasmodia were encapsulated by a granuloma comprising varying degrees of acute granulomatous inflammation. The new species is the fourth of Thelohanellus reported from North America and the first reported from Cyprinella, as well as the first myxozoan described from the blacktail shiner.
Anaplasmosis is caused by a Gram-negative obligate intracellular bacterium of the genus Anaplasma with the pathogen having a zoonotic impact. The study aimed to estimate the prevalence of anaplasmosis in Pakistan, to unravel the association of potential risk factors, and to investigate the effect on hematological parameters in affected small ruminants. A total of 150 (n = 75 sheep; n = 75 goats) blood samples were initially screened microscopically and then subjected to PCR targeting the amplification of the 16S rRNA gene fragment of Anaplasma. The PCR-based positive samples were then processed for sequencing. Statistical analysis regarding risk factors was performed using R software. The study revealed an overall 29.33% (44/150) prevalence of anaplasmosis in small ruminants. Sheep had higher (P > 0.05) prevalence (32%) as compared to goats (25.30%). The final statistical model resulting from backward elimination showed only tick infestation as a significant predictor of infection status. The phylogenetic analysis of 16S rRNA gene of Anaplasma spp. revealed 9 study isolates clustered together and showed a close resemblance (99%) with Anaplasma ovis isolate (DQ837600) from Hungary. One of the isolates showed (99%) similarity with the isolate of Anaplasma marginale (MH155594) from Iraq. Furthermore, the hematological parameters pack cell volume, red blood cells, hemoglobin, white blood cells, granulocytes, monocytes, lymphocytes, and platelet count were decreased in Anaplasma-positive animals. This is the first study at the molecular level to characterize Anaplasma spp. in small ruminants of Pakistan, and it will be useful in developing control strategies for anaplasmosis.
The laboratory BS-90 strain of the freshwater pulmonate snail Biomphalaria glabrata, progeny of snails collected from Salvador, Brazil, is resistant to infection with Schistosoma mansoni as juveniles or adults, which rapidly kill primary sporocysts with an attack by the internal defense system (IDS). However, neonatal snails are susceptible to infection. Although neonatal susceptibility of Salvador B. glabrata was reported in 1953 and confirmed subsequently, this phenomenon has been largely ignored. In this study, susceptibility was examined in discrete sizes (shell diameters) of BS-90 snails. We found that 1-mm snails are highly susceptible and develop patent infections. Unexpectedly, most infected 1-mm snails contain primary sporocysts in the digestive gland. Snails measuring 2 and 3 mm show a reduced prevalence of infection, and 4-mm and larger snails are refractory. In snails larger than 1 mm, sporocysts fail to develop normally, as shown by reduced numbers of germinal cells 48 hr post-exposure. Moreover, in larger snails an increasingly stronger response of the IDS is mounted in the form of increased numbers of sporocysts undergoing encapsulation and destruction by hemocytes, and increased layers of encapsulating hemocytes, as well as increased mitotic activity of the hematopoietic amebocyte-producing organ. These results indicate a relatively narrow size range over which resistance develops and suggest that the IDS of 1-mm snails is developmentally immature. The occurrence of infections in neonatal snails may help to explain the transmission of schistosomiasis in regions of low snail susceptibility and may complicate future efforts in biological control.
Heptacyclus buthi was harvested from fish hosts in rocky intertidal zones of Sonoma and Marin Counties, California, in October 2008 (n = 162) and October 2010 (n = 51). The size of the leeches was quantified using a method that approximated the sagittal cross-section of each specimen. Size-frequency curves were modeled to estimate the number of size-class cohorts in each year. If H. buthi is an annual species like many of its relatives, the single cohort modeled for in 2010 and the comparable “older” cohort in 2008, both with a broad range of sizes, may represent 1 component of its reproductive life history. A second, younger, more-numerous, less-variable cohort modeled from the 2008 sample may represent a second reproductive bout during that year that was prevented in the subsequent La Niña period of 2010–2011.
The long-term fidelity of pinniped hosts to their natal rookery site suggests the genetic architecture of their Uncinaria spp. hookworms should be strongly structured by host breeding biology. However, historical events affecting host populations may also shape parasite genetic structure. Sequences of the mitochondrial cytochrome c oxidase 1 (COI) gene of 86 Uncinaria lucasi individuals were obtained to assess genetic variation and structure of nematodes from 2 host species (68 hookworms from northern fur seals; 18 hookworms from Steller sea lions) and rookeries from 3 widely separated geographic regions: the western Bering Sea and Sea of Okhotsk, eastern Bering Sea, and the eastern Pacific Ocean. High COI haplotype (h = 0.96–0.98) and nucleotide (π = 0.014) diversity was found. The haplotype network showed a star-shaped pattern with a large number of haplotypes separated by few substitutions. The network did not show separation of U. lucasi by geographic region or host species. Fst values between U. lucasi individuals representing geographic regions showed no differentiation, consistent with the absence of genetic structure. At face value, this lack of genetic structure in U. lucasi suggests high gene flow but could also be explained by recent (post-glacial) population expansions of northern fur seals and their hookworms.
The objective of the study was to identify the seroprevalence of anti-Toxoplasma gondii antibodies in sheep herds from 3 municipalities from Jalisco, Mexico, as well as estimate the association between seroprevalence and certain factors presents in the farms. In total, 12 sheep farms that maintain only hair breeds were included in the work. From these farms, 336 blood samples were collected, corresponding 324 to ewes and 12 rams. Serum samples were subjected to ELISA test, and the association between the frequency of antibodies and some potential risk factors was estimated. The overall seroprevalence to anti-T. gondii antibodies in the population studied was 17.8% (60/336; 95% confidence interval [C.I.] 14–22), all farms had positive animals, and the seroprevalence of antibodies ranged between 7 to 32%. Seroprevalence in specific municipalities was 18.7% in Lagos de Moreno, 17.8% in Encarnación de Díaz, and 16.9% in San Juan de los Lagos. Seroprevalence in ewes was 17.5% (57/324; 95% C.I. 13–22), and seroprevalence in rams was 25% (3/12; 95% C.I. 6–57), while among breeds it was 17.8% in Pelibuey (20/112; 95% C.I. 11–26), 16.6% in Kathadin (14/ 84; 95% C.I. 9–26), 15.4% in Blackbelly (13/84; 95% C.I. 8–25), and 23.2% in Dorper (13/56; 95% C.I. 13–36); no differences were observed among breeds (p < 0.05). The presence of cats on the farms was associated with seroprevalence (odds ratio [OR] 2.8; 95% C.I. 1.8–7.3, p < 0.001), as was the absence of a rodent-control program (OR 1.5; 95% C.I. 0.8–3.2, p < 0.05). No other factors were identified as associated with seroprevalence.
The objective of the present study was to determine the characterization of Toxoplasma gondii in cats, rats, and chickens in the border areas of Yunnan Province. A total of 259 samples was collected from 10 border areas in Yunnan Province including 94 cats, 58 rats, and 107 chickens. Samples were screened by a nested polymerase chain reaction (PCR) assay and the positive products were analyzed by multilocus PCR-restriction fragment length polymorphism (RFLP) to determine the genotypes. Toxoplasma gondii deoxyribonucleic acid (DNA) was detected from 15.96% of 94 cats, 15.52% of 58 rats, and 6.54% of 107 chickens, respectively, and the average infection rate is 11.97%. Using the multilocus PCR-RFLP, we found that the genotype of T. gondii in cats and rats was ToxoDB#9. Because of low DNA concentration, no genotype was determined from chickens. These results fill the gaps of knowledge in the prevalence and genotype of T. gondii in the border areas of Yunnan Province and have implications for the better control of T. gondii infection in humans and animals.
Fatal infection by Cyathostoma (Cyathostoma) phenisci (Nematoda: Syngamidae), was identified in 2 of 52 brown boobies (Sula leucogaster) collected on beaches in the state of Rio de Janeiro, Brazil, and admitted to the veterinary clinic for rehabilitation. Both infected birds were in poor physical condition, with atrophied pectoral muscles, and died soon after starting treatment. The parasitological and pathological examination of the carcasses revealed the presence of C. (C.) phenisci in the trachea, resulting in tracheitis, as well as severe parasitic granulomatous bronchopneumonia caused by eggs deposited in the lungs. In our opinion, these serious pathological changes were the primary cause of chronic respiratory illness. This is the first description of fatal cyathostomiasis in a fish-eating avian host caused by infection by a member of the subgenus Cyathostoma (Cyathostoma). Therefore, it is reasonable to consider C. (C.) phenisci to be a real threat to a wide range of their definitive hosts, and cyathostomiasis should be considered in the differential diagnosis for fish-eating marine birds, even in cases without respiratory signs. This is also the first record of the genus Cyathostoma in Brazil.
New World flying squirrels, Glaucomys spp., are nocturnal arboreal sciurid rodents that have been previously surveyed for coccidial parasites. To date, 4 species of Eimeria have been reported from 2 species of Glaucomys. Here we report 2 species of eimerians from southern flying squirrels (Glaucomys volans) and the endemic Prince of Wales flying squirrel (Glaucomys sabrinus griseifrons). Oocysts of Eimeria dorneyi Levine and Ivens were found to be passing in the feces of 4 G. s. griseifrons from Alaska and a new species of Eimeria was present in feces from 6 G. volans from Arkansas. Oocysts of Eimeria hnidai n. sp. are ellipsoidal with a bilayered wall, measure 23.7 × 13.7 µm, and have a length/ width (L/W) ratio of 1.7; a micropyle and oocyst residuum are absent but polar granule(s) are present. Sporocysts are ellipsoidal–elongate and measure 11.8 × 4.9 µm, L/W 2.2; Stieda body is present but sub-Stieda and para-Stieda bodies are absent. The sporocyst residuum is composed of small indistinct granules along the edge or in the center of the sporocyst. This is the first coccidian reported from G. volans from Arkansas as well as the initial coccidian (E. dorneyi) reported from G. s. griseifrons from Alaska. We also provide a summation of the coccidia known from North American flying squirrels.
A survey of the parasite fauna of freshwater fishes from the Wet Tropics Bioregion in Queensland, Australia, revealed the presence of a new species of StemmatostomaCribb, 1986 (Digenea: Cryptogonimidae). Stemmatostoma cribbi n. sp. is described from the intestine and pyloric caeca of 2 species of grunter (Terapontidae), Hephaestus fuliginosus (Macleay) and Hephaestus tulliensis (De Vis), and the Jungle perch (Kuhliidae), Kuhlia rupestris (Lacepède), collected from the Barron and Mulgrave-Russell River drainage divisions in tropical north Queensland, Australia. Stemmatostoma cribbi is primarily distinguished morphologically from the type and only other species in the genus, Stemmatostoma pearsoniCribb, 1986, in having consistently fewer oral spines (14 in S. cribbi vs. 16 in S. pearsoni). Alignment of novel molecular data for S. cribbi and S. pearsoni revealed that they differ genetically by 26 nucleotides (2.1%) over the 1,258 bp partial large subunit (LSU) region, 1 nucleotide (0.8%) over the 121 bp partial 5.8S region, and 23 nucleotides (7.2%) over the entire 318 bp ITS2 rDNA region. Bayesian inference and maximum likelihood phylogenetic analyses of the partial LSU region for the species of Stemmatostoma sequenced here were used to explore the relationships of these species to other cryptogonimid species reported from freshwater ecosystems.
Of the 46 known species of Rhadinorhynchus Lühe, 1911, only 6 species, including Rhadinorhynchus dorsoventrospinosusAmin, Heckmann, and Ha, 2011, have dorsal and ventral, as well as lateral, trunk spines in the posterior field of trunk spines. The other 5 species are Rhadinorhynchus erumei Gupta and Fatima, 1981, Rhadinorhynchus adenati (Golvan and Houin, 1964) Golvan, 1969, Rhadinorhynchus lintoni Cable and Linderoth, 1963, Rhadinorhynchus pacificus Amin, Rubtsova, and Ha, 2019, and Rhadinorhynchus multispinosus Amin, Rubtsova, and Ha, 2019. These 5 species are distinguished from R. dorsoventrospinosus by differences in proboscis hook armature, trunk spine organization, and egg size. The distinction of R. dorsoventrospinosus is further demonstrated by its molecular description. We amplified the 18S and ITS1+5.8S+ITS2 rDNA region and cytochrome c oxidase subunit 1 (COI) gene for this study. Unfortunately, no ITS1+5.8S+ITS2 gene sequences are available for comparison with other species of the genus Rhadinorhynchus. Therefore, phylogenetic trees generated from sequences of the 18S nuclear region and COI gene were analyzed for the phylogenetic position of isolates of R. dorsoventrospinosus. Rhadinorhynchus dorsoventrospinosus has been validated as a species based on comparisons of morphological (original description) and molecular features (this paper). The additional genetic data will be useful as more species are described and as more genetic material becomes available to improve taxon sampling in the genetic analysis.
Commonly found in backyard and commercial poultry production, coccidiosis, caused by Eimeria species, presents a self-limiting intestinal infection based on the number of ingested oocysts. Heat stress (HS) is one of the major environmental stressors in poultry, predisposing broiler chickens to immunosuppression and rendering them susceptible to diseases. There are suggestions that HS reduces Eimeria oocyst shedding in chickens; however, the relationship between HS and coccidiosis is not well elucidated. The objective of this study was to investigate the effect of temperature on viability, morphology, infectivity, and development of Eimeria tenella in vitro, and merozoite production and oocyst shedding in vivo. In vitro exposure of sporozoites to 55 C for at least 60 min reduced sporozoites viability as shown by morphological changes and rendering them unable to invade Mardin-Darbi bovine kidney (MDBK) cells. Intracellular development of merozoites was significantly reduced by an increase in 2 C in the optimal temperature of incubation in vitro. Most importantly, the induction of HS in the live chickens caused significantly lower lesion scores, reduced merozoite production, and oocyst shedding, resulting in a much less severe disease outcome.
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