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17 recordsLinked to original sources

A comparative study of 28 culture media for Trichomonas gallinae

1. 1. A study was made of the ability of 28 different culture media to support growth of 5 strains of Trichomonas gallinae with their normally associated bacteria. A standard inoculum of 50 protozoa was used, and the cultures were incubated at 35 ?C. Based upon the number of positive cultures obtained, abundance of growth, and number of strains which grew in a given medium, the most satisfactory were Ringer-Loeffler serum, saline-Loeffler serum, and saline-serum. 2. 2. Pigeon serum used alone in a simple saline solution produced abundant growth and when added to other nutrients greatly enhanced the medium. Autoclaving of the serum appeared to have no effect on its growth promoting qualities. 3. 3. Neither egg yolk nor egg albumin alone appeared capable of supporting appreciable growth of T. gallinae. 4. 4. In general, the heavier the bacterial population supported by a medium the poorer the growth of T. gallinae. 5. 5. Strains of T. gallinae differ in their culturability. One strain grew in 82% of the media tested, another only in 43%.

Experimental Parasitology

A statistical analysis of the distribution of a larval nematode ( Anisakis sp. ) in the musculature of chum salmon ( Oncorhynchus keta - Walbaum)

The pepsin-HCl digestion technique is probably the best method of isolating Anisakis larvae from the musculature of chum salmon. Some losses can be expected due to breakage of the resistant cuticle of Anisakis , and can be estimated to be about 6% when counting the parasites with the unaided eye. Comparisons of Anisakis infections of the dorsal and ventral musculature indicated that very few larvae are found in the dorsal portions (above the lateral line). Forty-seven per cent of the fish had no dorsal infections, and 90% of the fish had less than 10% of total infection in the dorsal musculature. Comparisons of Anisakis infection ratios of the right and left sides below the lateral line showed some variation. Chi-square tests of the hypothesis that 50% of the infections occur in either side indicated no significant differences. There were indications that large total infection intensities displayed less variation. Comparisons of Anisakis infections of the anterior and posterior portions of the lateral musculature indicated that the infections were greater in the anterior portions. Calculated 95% confidence limits showed 50.0 to 59.0% infection in the anterior portion compared to 41.0 to 50.0% in the posterior portion. Analyses of the Anisakis distributions within the anterior portions of the lateral musculature were hindered by extreme variation in the counts. In general, an average of 60% of the anterior portion infections occurred in the upper two quadrants of musculature, and 40% in the lower two quadrants. Changes in distributional patterns were affected by total parasite density, rather than by age or geographical location. For purposes of comparing infection rates in chum salmon, distributional patterns indicate that the minimum sample of musculature which should be taken from each fish is one side below the lateral line. The one factor which probably contributes the greatest effect on distributional patterns of Anisakis within chum salmon musculature is the total intensity of infection (or population density of Anisakis ) in each fish.

Experimental Parasitology

Electron microscope study of Sarcocystis sp

Sarcocystis sp. obtained from wild populations of grackles, Quiscalus quiscula (Linn.), were examined to clarify the effect of the parasite on the host. Electron micrographs are presented to show areas of muscle destruction adjacent to the parasite which appear to be mechanically produced by the parasite. The microtubules within the villus-like projections of the cyst suggest that their possible function is absorptive and/or conductive with regard to the production of a toxin or the conveyance of nutritive material to the developing cells. The proposed function of submembranous filaments and their relation to the conoid is discussed. Similarities in the ultrastructure to Toxoplasma and other protozoa tend to negate the relegation of Sarcocystis to the fungi and further emphasize its protozoan nature.

Experimental Parasitology

How parasite exposure and time interact to determine Australapatemon burti (Trematoda: Digenea) infections in second intermediate hosts (Erpobdella microstoma) (Hirudinea: Erpodellidae)

Australapatemon spp. are cosmopolitan trematodes that infect freshwater snails, aquatic leeches, and birds. Despite their broad geographic distribution, relatively little is known about interactions between Australapatemon spp. and their leech hosts, particularly under experimental conditions and in natural settings. We used experimental exposures to determine how Australapatemon burti cercariae dosage (number administered to leech hosts, Erpobdella microstoma ) affected infection success (fraction to encyst as metacercariae), infection abundance, host survival, and host size over the 100 days following exposure. Interestingly, infection success was strongly density-dependent, such that there were no differences in metacercariae load even among hosts exposed to a 30-fold difference in cercariae. This relationship suggests that local processes (e.g., resource availability, interference competition, or host defenses) may play a strong role in parasite transmission. Our results also indicated that metacercariae did not become evident until ~4 weeks post exposure, with average load climbing until approximately 13 weeks. There was no evidence of metacercariae death or clearance over the census period. Parasite exposure had no detectable effects on leech size or survival, even with nearly 1,000 cercariae. Complementary surveys of leeches in California revealed that 11 of 14 ponds supported infection by A. burti (based on morphology and molecular sequencing), with an average prevalence of 32% and similar metacercariae intensity as in our experimental exposures. The extended development time and extreme density dependence of A. burti has implications for studying naturally occurring host populations, for which detected infections may represent only a fraction of cercariae to which animals have been exposed. Future investigation of these underlying mechanisms would be benefical in understanding host-parasite relationships.

Experimental Parasitology

Effects of malaria (Plasmodium relicturm) on activity budgets of experimentally-infected juvenile Apapane (Himatione sanquinea)

We used behavioral, physiological, and parasitological measures to document effects of acute malarial infections on activity budgets of experimentally infected juvenile Apapane ( Himatione sanguinea ). Five of eight birds died within 20 to 32 days after exposure to a single infective mosquito bite. Infected Apapane devoted less time to locomotory activities involving flight, walking or hopping, and stationary activities such as singing, preening, feeding, and probing. The amount of time spent sitting was positively correlated with parasitemia and increased dramatically after infection and between treatment and control groups. Birds that succumbed to infection experienced a significant loss of body mass and subcutaneous fat, whereas surviving Apapane were better able to maintain body condition and fat levels. When rechallenged with the parasite five months after initial infection, surviving birds experienced no increase in parasitemia, indicating that they had become immune to reinfection. Regardless of the outcome, infected birds experienced acute illness that would have left them unable to forage or to escape from predators in the wild.

Hawai'i

Effects of water temperature and substrate type on spore production and release in eastern Tubifex tubifex worms infected with Myxobolus cerebralis

Eastern Tubifex tubifex worms were exposed to Myxobolus cerebralis spores at 9, 13, 17, and 20 C in 1-L jars that contained sand, mud, or leaf litter as substrata. Beginning 60 days after exposure, water from each jar was filtered daily and examined for the presence of waterborne triactinomyxon spores (TAMs). On discovering a single TAM from an experimental jar, 48 T. tubifex worms from that jar were placed individually into 24-well plates. Spores released from individual infected T. tubifex worms were quantified to determine the first day of TAM release from infected worms, the infection rate, the total number of TAMs released per worm, and the duration of release. No TAMs were found in any of the jars incubated at 20 C or in uninfected, control worms at any temperature. The total number of TAMs released by infected worms in mud and sand was highest at 13 C compared with other temperatures. Infection rates among individual worms increased with temperature between 9 and 17 C. Higher temperatures (up to 17 C) induced earlier TAM releases among infected worms, and substratum did not influence this production parameter. The average duration of TAM release decreased as the temperature increased from 9 to 17 C, and there was a significant effect of substratum in the groups maintained at 13 and 17 C. In all temperature treatments between 9 and 17 C, the duration of release was least in the worms maintained in leaf litter, as was the total number of TAMs released during the experimental period and the median number of TAMs per production day.

Journal of Parasitology

Prairie dog responses to vector control and vaccination during an initial Yersinia pestis invasion

We evaluated the invasion of plague bacteria Yersinia pestis into a population of black-tailed prairie dogs (Cynomys ludovicianus; BTPDs) in South Dakota. We aimed to ascertain if Y. pestis invaded slowly or rapidly, and to determine if vector (flea) control or vaccination of BTPDs assisted in increasing survival rates. We sampled BTPDs in 2007 (before Y. pestis documentation), 2008 (year of confirmed invasion), and 2009 (after invasion). We estimated annual BTPD re-encounter rates on three 9-ha plots treated annually with deltamethrin dust for flea control and three 9-ha plots lacking dust. In 2007 and 2008, approximately half the adult BTPDs live-trapped were injected subcutaneously with either an experimental plague vaccine (F1–V fusion protein) or placebo formulation; the remaining individuals were not inoculated. From 2007 to 2009, we sampled 1559 BTPDs on 2542 occasions. During 2007–2008, the prevalence and intensity of fleas on BTPDs were 69–97% lower on the dusted vs. no dust plots. From 2007 to 2008, the annual re-encounter rate of non-inoculated BTPDs was 150% higher on the dusted vs. no dust plots. During the same interval on the dusted plots, the re-encounter rate was 55% higher for vaccinated adult female BTPDs vs. nonvaccinated adult females, but the annual re-encounter rate was 19% lower for vaccinated adult males. By late August 2008, BTPDs were nearly extirpated from the no dust plots. During 2007–2008 and 2008–2009 on the dusted plots, which persisted, the BTPD re-encounter rate was 41% higher for vaccinated vs. non-vaccinated adult females but 35% lower for vaccinated adult males. Yersinia pestis erupted with vigor as it invaded. Flea control enhanced BTPD survival but did not offer full protection. Flea control and F1–V vaccination seemed to have additive, positive effects on adult females. Annual re-encounter rates were reduced for vaccinated adult males; additional experimentation is needed to further evaluate this trend.

International Journal of Parasitology: Parasites a

Diagnostic methodology is critical for accurately determining the prevalence of ichthyophonus infections in wild fish populations

Several different techniques have been employed to detect and identify Ichthyophonus spp. in infected fish hosts; these include macroscopic observation, microscopic examination of tissue squashes, histological evaluation, in vitro culture, and molecular techniques. Examination of the peer-reviewed literature revealed that when more than 1 diagnostic method is used, they often result in significantly different results; for example, when in vitro culture was used to identify infected trout in an experimentally exposed population, 98.7% of infected trout were detected, but when standard histology was used to confirm known infected tissues from wild salmon, it detected ~50% of low-intensity infections and ~85% of high-intensity infections. Other studies on different species reported similar differences. When we examined a possible mechanism to explain the disparity between different diagnostic techniques, we observed non-random distribution of the parasite in 3-dimensionally visualized tissue sections from infected hosts, thus providing a possible explanation for the different sensitivities of commonly used diagnostic techniques. Based on experimental evidence and a review of the peer-reviewed literature, we have concluded that in vitro culture is currently the most accurate diagnostic technique for determining infection prevalence of Ichthyophonus, particularly when the exposure history of the population is not known.

Journal of Parasitology

Evidence of experimental postcyclic transmission of Bothriocephalus acheilognathi in bonytail chub (Gila elegans)

We examined the role that predation of infected conspecific fish and postcyclic transmission might play in the life cycle of the Asian fish tapeworm, Bothriocephalus acheilognathi (Cestoda: Pseudophyllidea) Yamaguti, 1934. Young-of-the-year (YOY) bonytail chub ( Gila elegans ) were exposed to copepods infected with B. acheilognathi and subsequently fed to subadult bonytail chub. Within 1 wk after consumption of the YOY chub, subadults were necropsied and found infected with gravid and nongravid tapeworms. This study provides evidence that postcyclic transfer of B. acheilognathi can occur. Postcyclic transmission may be an important life history trait of B. acheilognathi that merits consideration when studying the impact and distribution of this invasive and potentially pathogenic tapeworm.

New Mexico

Sarcocystis neurona infections in sea otter (Enhydra lutris): Evidence for natural infections with sarcocysts and transmission of infection to opossums (Didelphis virginiana)

Although Sarcocystis neurona has been identified in an array of terrestrial vertebrates, recent recognition of its capacity to infect marine mammals was unexpected. Here, sarcocysts from 2 naturally infected sea otters (Enhydra lutris) were characterized biologically, ultrastructurally, and genetically. DNA was extracted from frozen muscle of the first of these sea otters and was characterized as S. neurona by polymerase chain reation (PCR) amplification followed by restriction fragment length polymorphism analysis and sequencing. Sarcocysts from sea otter no. 1 were up to 350 I?m long, and the villar protrusions on the sarcocyst wall were up to 1.3 I?m long and up to 0.25 I?m wide. The villar protrusions were tapered towards the villar tip. Ultrastructurally, sarcocysts were similar to S. neurona sarcocysts from the muscles of cats experimentally infected with S. neurona sporocysts. Skeletal muscles from a second sea otter failed to support PCR amplification of markers considered diagnostic for S. neurona but did induce the shedding of sporocysts when fed to a laboratory-raised opossum (Didelphis virginiana). Such sporocysts were subsequently fed to knockout mice for the interferon-gamma gene, resulting in infections with an agent identified as S. neurona on the basis of immunohistochemistry, serum antibodies, and diagnostic sequence detection. Thus, sea otters exposed to S. neurona may support the development of mature sarcocysts that are infectious to competent definitive hosts.

Washington

PCR diagnostics underestimate the prevalence of avian malaria ( Plasmodium relictum ) in experimentally-infected passerines

Several polymerase chain reaction (PCR)-based methods have recently been developed for diagnosing malarial infections in both birds and reptiles, but a critical evaluation of their sensitivity in experimentally-infected hosts has not been done. This study compares the sensitivity of several PCR-based methods for diagnosing avian malaria ( Plasmodium relictum ) in captive Hawaiian honeycreepers using microscopy and a recently developed immunoblotting technique. Sequential blood samples were collected over periods of up to 4.4 yr after experimental infection and rechallenge to determine both the duration and detectability of chronic infections. Two new nested PCR approaches for detecting circulating parasites based on P. relictum 18S rRNA genes and the thrombospondin-related anonymous protein (TRAP) gene are described. The blood smear and the PCR tests were less sensitive than serological methods for detecting chronic malarial infections. Individually, none of the diagnostic methods was 100% accurate in detecting subpatent infections, although serological methods were significantly more sensitive (97%) than either nested PCR (61–84%) or microscopy (27%). Circulating parasites in chronically infected birds either disappear completely from circulation or to drop to intensities below detectability by nested PCR. Thus, the use of PCR as a sole means of detection of circulating parasites may significantly underestimate true prevalence.

Journal of Parasitology

An experimental evaluation of host specificity: The role of encounter and compatibility filters for a rhizocephalan parasite of crabs

The encounter/compatibility paradigm of host specificity provides three qualitative pathways to the success or failure of a potential host-parasite interaction. It is usually impossible to distinguish between two of these (encounter and compatibility filters closed versus encounter filter open and compatibility filter closed) because unsuccessful infection attempts are difficult to observe in nature. We were able to open the encounter filter under experimental laboratory conditions. Our analytical system used the rhizocephalan barnacle, Sacculina carcini , a parasitic castrator of the European green crab, Carcinus maenas , and Pachygrapsus marmoratus , a native European crab that occurs with C. maenas but is not parasitized by S. carcini in nature. Penetration followed by unsuccessful infection of P. marmoratus crabs by parasitic barnacle larvae leaves a uniquely permanent record in the thoracic ganglion of the crabs. This provided us with a novel tool to quantify the encounter filter in a host-parasite system in nature. We demonstrated, in the laboratory, that the compatibility filter was closed and that, in nature, even where barnacle larvae were present, the encounter filter was also effectively closed. The closure of both filters in nature explains the failure of this potential host-parasite interaction, an outcome favored by selection in both host and parasite.

International Journal for Parasitology

Relationship of tadpole stage to location of echinostome cercaria encystment and the consequences for tadpole survival

The effect of echinostome infections on the survival of Rana pipiens tadpoles was examined in relation to developmental stage of tadpoles. Individual tadpoles of Gosner stages 25, 27, 32–33, and 37–39 were exposed to 1 of 4 levels of cercariae (0, 20, 50, or 100). Only tadpoles at stage 25, the earliest stage infected, died within a 5-day experimental period. This stage-specific mortality rate could be explained, in part, by the stage-specific location of encystment of cercariae, which was documented in a separate experiment. In accordance with kidney development, cercariae predominately encysted in the pronephroi during early stages of tadpole development (stages 25 through 31–32) and only in the mesonephroi and associated ducts at later stages (stages 37 through 46). As the mesonephros develops, renal capacity presumably increases. Thus, tadpoles died only when metacercariae concentrated in the functional portion of the kidney with the most limited renal capacity. As tadpoles aged, they also became less susceptible to infections. On average, 69.5% of cercariae that were exposed to stage 25–26 tadpoles successfully encysted, compared with only 8.4% of cercariae exposed to stage 37–38 tadpoles. Exposures of metamorphic frogs (poststage 46) to cercariae revealed that these individuals can become infected with echinostomes. Collectively, our data highlight the host stage–dependent dynamics of tadpole–echinostome interactions.

Journal of Parasitology

Wildlife disease and conservation in Hawaii: pathogenicity of avian malaria (Plasmodium relictum) in experimentally infected Iiwi (Vestiaria coccinea)

Native Hawaiian forest birds are facing a major extinction crisis with more than 75% of species recorded in historical times either extinct or endangered. Reasons for this catastrophe include habitat destruction, competition with non-native species, and introduction of predators and avian diseases. We tested susceptibility of Iiwi (Vestiaria coccinea), a declining native species, and Nutmeg Mannikins (Lonchura punctulata), a common non-native species, to an isolate of Plasmodium relictum from the island of Hawaii. Food consumption, weight, and parasitaemia were monitored in juvenile Iiwi that were infected by either single (low-dose) or multiple (high-dose) mosquito bites. Mortality in both groups was significantly higher than in uninfected controls, reaching 100% of high-dose birds and 90% of low-dose birds. Significant declines in food consumption and a corresponding loss of body weight occurred in malaria-infected birds. Both sex and body weight had significant effects on survival time, with males more susceptible than females and birds with low initial weights more susceptible than those with higher initial weights. Gross and microscopic lesions in malaria fatalities included massive enlargement of the spleen and liver, hyperplasia of the reticuloendothelial system with extensive deposition of malarial pigment, and overwhelming anaemia in which over 30% of the circulating erythrocytes were parasitized. Nutmeg Mannikins, by contrast, were completely refractory to infection. Our findings support previous studies documenting high susceptibility of native Hawaiian forest birds to avian malaria. This disease continues to threaten remaining high elevation populations of endangered native birds.

Hawai'i

Life cycle of the trout cecal nematode, Truttaedacnitis truttae (Nematoda: Cucullanidae): Experimental and field observations

Truttaedacnitis truttae is a cucullanid nematode of primarily salmonine fishes. Brown trout ( Salmo trutta ) in Europe reportedly become parasitized by ingesting lampreys ( Lampetra planeri ) carrying infective larvae. However, our field and laboratory observations suggested that North American specimens of T. truttae have an alternative life cycle. High abundances and potential impact of T. truttae in rainbow trout, Oncorhynchus mykiss , in the Colorado River drainage in Grand Canyon, where there are no lampreys, prompted a study on the transmission dynamics of this nematode. Eggs of T. truttae , collected from live gravid females, were incubated in the laboratory. Snails, Physa gyrina and Lymnaea sp., were exposed to T. truttae larvae 3–4 wk later. Active larvae of T. truttae were observed penetrating the intestinal wall of exposed snails, and worm larvae were found in the visceral tissues when examined 1 wk after exposure. Larvae in snails showed little growth and development 2 wk later and corresponded to L3 larvae. Infected snails were fed to hatchery-reared juvenile rainbow trout. Developing stages were subsequently found in the mucosal lining and lumen of trout intestines. Adult male and female (gravid) worms were found in the ceca of trout examined 5–6 mo after consuming infected snails. Larvae found in pepsin/trypsin digests and mucosal scrapings from wild, naturally infected, trout corroborate laboratory findings. Screening of Physa sp. and gammarids collected from Colorado River, Grand Canyon, for natural infections with T. truttae using the ITS1 rDNA marker gave positive results. Truttaedacnitis truttae is the second species, after Truttaedacnitis clitellarius of lake sturgeon, capable of using a snail first intermediate/paratenic host and is similar to several other cucullanids in having a histotropic phase of development in the definitive fish host.

Arizona