Showing papers in "Journal of Parasitology in 1992"

Isoenzyme analysis of 35 Toxoplasma gondii isolates and the biological and epidemiological implications.

Abstract: Isoenzyme analysis was conducted on the tachyzoite stage of 35 Toxoplasma gondii isolates. Fifteen enzyme systems were studied after isoelectrofocusing of tachyzoite extracts in polyacrylamide or agarose gels. Six enzyme systems showed variable electrophoretic patterns: aspartate aminotransferase (EC 2.6.1.1), glutathione reductase (EC 1.6.4.2), glucose phosphate isomerase (EC 5.3.1.9), amylase (EC 3.2.1.1), acid phosphatase (EC 3.1.3.2), and propionyl esterase. Their combination allows the description of 5 zymodemes among the 35 T. gondii isolates. Zymodeme 1 involves 6 isolates that are highly pathogenic to mice and for which oocysts could not be obtained. Isolates belonging to zymodemes 2, 3, and 4 are less pathogenic to mice and produced oocysts. Zymodeme 5 involves only 1 isolate, which was highly pathogenic to mice.

Taxonomic revision of the genus Trichinella.

Abstract: The analysis of genetic, biochemical, and biological data on about 300 Trichinella isolates, reported in the literature, allows a taxonomic revision of this genus. We propose the recognition of 5 sibling species, Trichinella spiralis (Owen, 1835) sensu stricto; Trichinella nativa Britov and Boev, 1972; Trichinella pseudospiralis Garkavi, 1972; Trichinella nelsoni Britov and Boev, 1972 sensu stricto; and Trichinella britovi n. sp., on the basis of biochemical and biological characteristics. Trichinella britovi n. sp. is characterized by distribution in the Palaearctic Region; newborn larvae (NBL) production in vitro of 35-55 NBL/72 hr; nurse cell development time (NC d.t.) between 24 and 42 days postinfection (d.p.i.); low reproductive capacity index (RCI) in mice, rats, and pigs; low resistance to freezing; 1 unique marker allozyme; and moderate pathogenicity for humans. The new species is most similar to Trichinella nativa but differs from it in 4 allozymes, in having less resistance to freezing, in having a different pattern of major ribosomal DNA fragments after endonuclease digestion, and in distribution area. Trichinella nativa is characterized by a holarctic distribution; hosts that are sylvatic mammals; NBL production in vitro 28-54/72 hr; NC d.t. between 20 and 30 d.p.i.; low RCI in mice, rats, and pigs; high resistance to freezing; 2 unique marker allozymes; and moderate to severe pathogenicity for humans. Trichinella spiralis sensu stricto is characterized by a cosmopolitan distribution in domestic pigs, associated wildlife, and humans; high NBL production in vitro (greater than 90 NBL/72 hr); NC d.t. between 16 and 37 d.p.i.; high RCI in mice, rats, and pigs; no resistance to freezing; 6 unique marker allozymes; and high pathogenicity for humans.(ABSTRACT TRUNCATED AT 250 WORDS)

Population dynamics of Plasmodium falciparum sporogony in laboratory-infected Anopheles gambiae.

Abstract: The population dynamics of cultured Plasmodium falciparum parasites was examined during their sporogonic development in Anopheles gambiae mosquitoes. Estimates of absolute densities were determined for each life stage, and life tables were constructed for each of 38 experimental infections. Macrogametocyte and ookinete mortalities contributed equally to the overall mortality. On average, there was a 40-fold decrease in parasite numbers in the transition from the macrogametocyte to the ookinete stage, a 69-fold decrease in the transition from ookinete to oocyst stages, and a total net decrease in parasite numbers from macrogametocyte to oocyst stage of 2,754-fold (i.e., multiplicative). There was no relationship between macrogametocyte and ookinete densities due to the inherent variability in fertility among different gametocyte cultures. There was a curvilinear relationship (r2 = 0.66) between ookinete and oocyst densities. Above a threshold of about 30 ookinetes/mosquito, the oocyst yield per ookinete became increasingly greater with increasing ookinete density. There was a linear relationship (r2 = 0.73) between oocyst and sporozoite densities, with an average of 663 salivary gland sporozoites produced per oocyst. Sporozoite production per oocyst was not affected by oocyst density and virtually all oocyst infections resulted in sporozoite infections of the salivery glands. This quantitative study indicates that the sporogony of cultured P. falciparum in laboratory-infected A. gambiae is an inefficient process and that the ookinete is the key transitional stage affecting the probability of vector infectivity.

Biological characterization of Trichinella isolates from various host species and geographical regions.

Abstract: Forty isolates of Trichinella collected from 5 continents were compared for 7 biological characters: newborn larvae produced per female worm cultured in vitro at the seventh, eighth, and ninth day postinfection, host muscle nurse cell development time, reproductive capacity index in rats and chickens, and resistance of muscle larvae to freezing. The isolates also were compared by analyses of an environmental character of the location from which they were isolated: the isotherms for January and July. By factorial analysis of correspondence of the biological and environmental data, the 40 isolates were grouped into 8 gene pools (T1-T8). The environmental temperature-related distribution was more evident for the sylvatic isolates (T2, T3, T5, T6, T7, T8), than for T1, which was isolated from domestic pigs, and for T4, a bird-adapted, nonencapsulating genetic type. The 8 biological groups correlated closely with the 8 gene pools previously identified on the basis of allozyme analysis. These results support the concept that the genus Trichinella is composed of at least 5 distinct gene pools or sibling species: Trichinella spiralis sensu stricto (T1), Trichinella nativa (T2), Trichinella sp. (T3), Trichinella pseudospiralis (T4), and Trichinella nelsoni (T7), and 3 other groups of uncertain taxonomic status (i.e., T5, T6, and T8).

The role of the mosquito peritrophic membrane in bloodmeal digestion and infectivity of Plasmodium species.

Abstract: Secretion and luminal formation of the peritrophic membrane (PM) were induced in female Anopheles stephensi and Aedes aegypti by feeding the mosquitoes on a warmed suspension of latex particles in Ringer's solution. The PM in A. stephensi was produced from apical secretion vesicles stored in the midgut epithelial cells and secreted into the lumen during feeding. In A. aegypti, the PM was formed de novo. When the latex feeding was followed 24 hr later by a meal of lyophilized pig blood, the 2 mosquito species exhibited very different modifications to their PM structure; in A. stephensi no PM was formed around the blood meal, whereas de novo synthesis of the PM in A. aegypti continued during the blood meal, with the resulting PM greatly thickened compared to the normal feeding. This artificial induction of PM formation was used as the basis to study the role of the PM in blood meal digestion and in infectivity of mosquitoes by the appropriate species of Plasmodium. The feeding of a latex suspension alone had no stimulatory effect on the 2 major midgut proteases, trypsin and aminopeptidase, in either species. After a blood meal alone, proteases rose to maximum activity at 30 hr and 24 hr after feeding in A. stephensi and A. aegypti, respectively. After double feeding, protease activities in both species were almost identical to those in blood-fed mosquitoes. Neither the absence of a PM (in A. stephensi) nor the presence of a thickened PM (in A. aegypti), therefore, has any effect on the ability of mosquitoes to digest a blood meal. Malaria infectivity, measured by oocyst counts, also was compared after normal and double feeding using infective blood meals. Infectivity of A. stephensi by Plasmodium berghei was unaffected by the presence or absence of the PM. The thickened PM produced by double feeding in A. aegypti caused a reduction of midgut infectivity by Plasmodium gallinaceum. These results suggest that the PM may act as a partial, but not an absolute, barrier to invasion of the midgut by the ookinete.

Infectivity of plerocercoids of Schistocephalus solidus (Cestoda: Ligulidae) and fecundity of the adults in an experimental definitive host, Gallus gallus.

Abstract: Aspects of the infectivity of the plerocercoid stage and the fecundity of the adult stage of Schistocephalus solidus were examined using the chicken, Gallus gallus, as an experimental host. To investigate size-related infectivity of the plerocercoid stage to a definitive host, a range of plerocercoids (166) were weighed, and each was fed to an individual male chicken. Only a very small percentage of plerocercoids weighing less than 50 mg established compared with in excess of 50% in all other weight classes. To examine the factors affecting the quantity of eggs produced by the adult stage, 15 similar-sized plerocercoids (160-218 mg) were fed to chickens. The majority of plerocercoids administered established as adults and survived until the experiment was terminated on day 7 postinfection (PI). Fecal egg counts indicated that all established worms commenced egg production on day 2 PI with peak output on day 2 or 3 PI. By day 7 PI, egg production in the surviving worms had declined, but it was still evident. The final weight of the adult and the average egg output per worm were unrelated to the initial weight of the infecting plerocercoid. However, the average egg output was predicted by the final adult dry weight and to a lesser extent by the proportion of weight lost in the transition from plerocercoid to adult, but not by the absolute weight loss.

Behavioral adaptations to parasites: an ethological approach.

Abstract: Wild vertebrate animals must live in an environment with the ever present threat of internal and external parasites. This threat by macroparasites is responsible for the natural selection of an array of behavioral adaptations that, together with the immune system and other physiological forms of resistance, enable the animals to survive and reproduce in this environment. Several lines of research, some quite recent, illustrate that specific behavioral patterns can be effective in helping animals or their offspring avoid or control macro- parasites that can affect adversely the animal's fitness. These behavioral patterns fall under the general strategies of avoidance behavior and mate selection. Various aspects of social, feeding, and repro- ductive behavior of vertebrates are shaped by the forces of predation and resource limitation that animals must address to survive to repro- ductive age and successfully rear young. A rel- atively unappreciated force shaping behavior in much this same way is the existence of external and internal parasites. Animals that are studied by scientists usually are maintained in relatively clean laboratories, field stations, and domestic environments, where it is easy to forget that the animals evolved and thrived in environments with an array of parasites long before human protective measures were available. The behav-

Allozyme analysis of Trichinella isolates from various host species and geographical regions.

Abstract: Allozyme analysis was carried out on 152 Trichinella isolates from synanthropic and wild animals and from humans; the isolates were collected from 5 continents. The analysis, involving 27 enzymes, revealed the presence of 8 distinct gene pools, termed T1-T8. Four of the genetic groups represent the 4 previously proposed species: Trichinella spiralis sensu stricto (T1), Trichinella nativa (T2), Trichinella nelsoni (T7), and Trichinella pseudospiralis (T4). The other 4, T3, T5, T6, and T8 are distinct from previously described species. The absence of allozymic hybrid patterns among even sympatric groups indicates a lack of gene flow among the groups. Principal component analysis and the unweighted pair group method of analysis were used to assemble allozyme patterns of the 152 isolates into discrete groups and to show their relative relationships. Both analyses indicated the presence of 8 primary clusters that correlated with the gene pools revealed by direct allozyme profile analysis. The absence of evidence of gene flow among the gene pools and the high level of allozymic differentiation between the cluster groups support the concept that the genus Trichinella is composed of several sibling species.

Coevolution: a history of the macroevolutionary approach to studying host-parasite associations.

Abstract: The year 1991 marked 100 yr of coevolution research. I have reviewed the first 90 yr of this history. Three chronological phases are apparent: recognition of predictable associations among hosts and their parasites; search for patterns of association and their underlying causes, emphasizing either correlated biogeographic patterns or correlated phylogenies; and development of objective and repeatable methodologies for reconstructing and interpreting these patterns of association. Von Ihering, an outspoken anti-Darwinian, was undoubtedly the first to recognize and make use of predictable host-parasite associations. Kellogg and Fahrenholz, however, had more profound influence on subsequent generations, but in different directions. Kellogg attempted to meld natural selection with speciation by isolation. He also considered host specificity a component of coevolution, important but variable. His work laid the foundation for future research concentrated on biogeographic interpretations of host-parasite relationships. This emphasis and Metcalf's failed attempts to provide adequate mechanisms for reconstructing phylogenies reduced the biogeographic approach to an empirical research program in the hands of Manter. Fahrenholz, on the other hand, exposed to a strong anti-Darwinian sentiment, emphasized the importance of strict host specificity. This led to Eichler's formulation of the first 3 coevolutionary rules and the conclusion that host specificity was not a component but the cause of coevolution and ultimately the tautology inherent in the phylogenetic approach. All had to rely on 1 assumption, that host and parasite phylogenies were reflected in the taxonomic hierarchy. Hennig criticized this assumption and provided a method whereby phylogenies are reconstructed independently. Brooks melded this new phylogenetic method (cladistics) with an equally new biogeographic method (vicariance biogeography), providing the foundations for the modern macroevolutionary approach to studying host-parasite associations.

Toxoplasma gondii: differentiation and death of bradyzoites.

Abstract: The living parasites inToxoplasma cysts cannot be eradicated by current therapy and maintain latent infections for many years. Relatively little is known about encystedToxoplasma. We therefore undertook studies using mice infected with the avirulent ME 49 strain ofToxoplasma. The bradyzoites in young (12- to 17-day-old) cysts contained the same organelles as did tachyzoites. The bradyzoites of older cysts (4 weeks postinoculation) had differentiated, losing certain organelles and acquiring others. Our major new finding was that in animals inoculated ≥4 weeks previously, some bradyzoites were totally disrupted, splling their contents (perhaps including lytic substances) into the cyst matrix. Many older bradyzoites in the same cysts lacked internal membranes and their viability was questionable, but there were also occasional parasites resembling viable tachyzoites and mature bradyzoites, organisms that might possibly initiate daughter cyst formation after cyst rupture. The life span of an individual bradyzoite may be shorter than formerly appreciated despite the prolonged course of latent infections.

Physiological analysis of cercarial behavior.

Abstract: The behavior of trematode cercariae was made accessible to physiological analyses by splitting the continuous flow of behavior into separate units of behavior patterns. Such phases include active, passive, and resting phases in the intermittent swimming mode, attachment to the host, remaining on the host, directed creeping to suitable entry sites, and penetration phases including penetration movements, tail shedding, tegument transformation, and secretion of enzymes. Each of these phases may be stimulated by separate environmental and host signals. The pattern of the responses and the chemical, thermal, mechanical, and visual stimuli have been described in some detail but only a few studies have dealt with the question of how these responses are coordinated by receptors and nervous systems. Highly specific and sensitive chemoreceptors for host signals such as carbon dioxide, L-arginine, fatty acids, and glycoproteins have been defined from cercarial behavior, but they have not yet been allocated to morphological structures. Analyses of cercarial behavior of 6 schisto- somatid and 4 fish-infecting species revealed that the individual parasite species show different behavioral patterns and respond to very different host signals, even though they infect congeneric hosts. The adaptive benefits of such behavioral diversity and complexity still are to be elucidated. The complex life cycles of trematodes include very different methods of distribution and trans- fer from one host to another. The behavior of

Neospora caninum (Apicomplexa) in a stillborn goat.

Abstract: Tissue cysts of Neospora caninum were found in sections of brain from a stillborn pygmy goat. The tissue cysts had 1-2-microns-thick cyst walls and stained with anti-N. caninum serum in an immunohistochemical test. Glial nodules, mononuclear perivascular cuffing, and foci of inflammation were associated with N. caninum tissue cysts throughout the brain. This report indicates that N. caninum can be transmitted transplacentally in goats.

Parasites as probes for biodiversity.

Abstract: Cestodes of the genus Linstowia, parasitic in marsupials, show patterns of coevolution and ancient historical-ecological connections. Correlated with the breakup of the austral landmasses (Gondwanaland) of the Neotropical and Australian regions from the Antarctic continent, the age of this host-parasite community is estimated to be between 60 and 70 million years old. Based on the data from the survey of parasites of mammals from throughout Bolivia and from the phylogenetic analysis of the cestodes, we urge the planners of biodiversity preserves in the neotropics to consider the Yungas of Bolivia as a region that supports an ancient ecological community worthy of consideration as a biopreserve.

Excretory-secretory products of Echinostoma paraensei sporocysts mediate interference with Biomphalaria glabrata hemocyte functions.

Abstract: Miracidia of Echinostoma paraensei were cultured in medium containing 14C-labeled amino acids, allowed to transform into sporocysts, and their excretory/secretory products (E-S) were collected and characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis and autoradiography. Effects of E-S on hemocytes of Biomphalaria glabrata were also assessed. E-S collected during day 1 of culture (E-S1) contained several polypeptides, none of which were labeled, suggesting that E-S1 are largely preformed. E-S1 significantly depressed the ability of hemocytes to phagocytose sheep red blood cells (SRBC), but otherwise had little effect on hemocyte structure or behavior. E-S released by sporocysts in day-2 cultures (E-S2) and in older cultures generally were similar and also contained several polypeptides, many of which were labeled, indicating active synthesis of E-S in vitro. E-S2 strongly inhibited hemocyte uptake of SRBC. Also, hemocytes pretreated with E-S2 assumed a spherical shape and failed to spread normally. E-S obtained through 10 days of culture mediated this effect. Active components of E-S2 were greater than 100 kDa in their native configuration, were heat- and trypsin-labile, and were bound by anti-E-S antibodies. Both greater than 200- and 80-kDa bands were prominent in anti-E-S immunoprecipitates. Hemocytes derived from snails of the 13-16-R1 strain of B. glabrata (a strain resistant to infection with Schistosoma mansoni), when pretreated with E-S2, bound to sporocysts of S. mansoni but lost their ability to damage such sporocysts. E-S2 interfered with hemocyte functions in ways inferred from earlier classic in vivo studies of trematode-snail interactions.

The nervous systems of helminths as targets for drugs.

Abstract: Processes that critically differentiate parasitic helminths and their hosts are obvious candidates for chemotherapeutic intervention. The recognition that neurobiology distinguishes helminths from their vertebrate hosts is due in part to the fact that several efficacious anthelmintics, derived generally from empirical screening, have been found to act selectively on the neuromuscular system of these parasites. In addition, basic physiological and pharmacological research has revealed considerable differences in the ways in which helminths and their hosts transmit information in the nervous system and respond to it in innervated tissues. Unfortunately, most of these differences have yet to be exploited in chemotherapy. The topics for this review include an analysis of mechanistic aspects of the pharmacology of anthelmintics that act on neuromuscular systems and a consideration of the prospects for discovery of novel drugs that act on this system.

Equine protozoal myelitis in Panamanian horses and isolation of Sarcocystis neurona.

Abstract: Schizonts of Sarcocystis neurona were identified microscopically in hematoxylin-eosin-stained spinal cord sections from 2 native Panamanian horses that exhibited clinical signs of equine protozoal myelitis (EPM). Spinal cord homogenate from a third Panamanian horse with EPM was inoculated onto monolayers of cultured bovine monocytes (M617). Intracytoplasmic schizonts containing merozoites arranged in rosette forms surrounding a central residual body first were observed 13 wk postinoculation. Parasites divided by endopolygeny and lacked rhoptries. Schizonts from each horse reacted with Sarcocystis cruzi antiserum in an immunohistochemical test.

The roles of ecological and evolutionary influences in providing structure to parasite species assemblages.

Abstract: Parasite species assemblages currently are thought to range from isolationist to interactive, their dynamic properties being related to the number of species and types of hosts involved. The literature contains few experimental tests of this concept, however, and many of the host/parasite systems studied to date are not amenable to experimental manipulation. In this review, the presence of a parasite species, in a sample of host individuals, is considered to be an evolutionary phenomenon, but the parasite's population structure is considered to be an ecological one. Studies that allow evaluation of these 2 influences are comparative in nature and include data from a series of homogeneous samples of host populations. A lottery model is presented, in which hosts acquire their assemblages of parasites by Monte Carlo type sampling from multiple kind arrays; the major structuring influence is the relative probability of becoming infected by various parasite species. Claims of parasite species interaction need to be supported by studies showing departures from the predictions of this model. The species density and infraassemblage diversity index distributions are recommended as quantitative

Immunological mediation of gonadal effects on experimental murine cysticercosis caused by Taenia crassiceps metacestodes

Abstract: Female BALB/c mice are naturally more susceptible than males to intraperitoneal experimental infection with Taenia crassiceps metacestodes. Gonadectomy tends to equalize susceptibility between sexes by reducing in half the mean individual intensity of females and by tripling that of males. The effect of gonadectomy is seen only in mice with intact immune systems but not in irradiated mice. Purified sex hormones (17-# estradiol, testosterone, and progesterone) do not affect cysticercus reproduction or growth in vitro. Thus, gonadal effect on mouse susceptibility to cysticercosis appears to be mediated via the immune system, and it is probably not the consequence of the major sex steroids acting directly upon the parasites. Because sublethal irradiation increases the intensity in gonadectomized females and intact males, whereas that of gonadectomized males and intact females remains unchanged, irradiation results are consistent with the hypothesis that immunological events that participate in controlling the growth of cysticerci are inhibited by ovaries and stimulated by testes. Experimental murine cysticercosis caused by Taenia crassiceps metacestodes greatly facilitates the study of biological factors participating in metacestode disease because T. crassiceps cys- ticerci grow quickly within the mouse intraperi- toneal cavity and are kept easily in vitro. The structural, morphological, and immunological similarities shared by T. crassiceps with other cestodes may relate experimental results to other infections including those of humans (Larralde et al., 1988, 1990). Previous studies on murine

Isolation of Toxoplasma gondii from a naturally infected beef cow.

Abstract: Toxoplasma gondii was isolated from the intestinal wall of an adult shorthorn cow. The cow had an antibody titer of 1:1,600 in the T. gondii agglutination test using formalin-fixed whole tachyzoites. The strain of the parasite designated CT-1 was lethal to mice. This is the first report of the recovery of viable T. gondii from a naturally infected cow in the United States.

Congruent and synchronic patterns in biogeography and speciation among seabirds, pinnipeds, and cestodes.

Abstract: Congruence in biogeographic patterns among diverse assemblages of taxa indicates uniformity in the historical determinants of biotic distributions. Comparisons of host and parasite phylogenies and the elucidation of distributional area relationships are requisite components of analyses in historical biogeography. Host-parasite associations with broad geographic ranges are often archaic and have been structured largely by coevolutionary processes. In contrast, the origins and radiation of the primary cestode faunas of some seabirds (Alcataenia spp./Alcidae) and pinnipeds (Anophryocephalus spp./Phocidae and Otariidae) are associated with colonization. These young colonizing faunas, in the Holarctic Region, were influenced by a common history during the late Pliocene and Pleistocene epochs. Periodic range contraction, with isolation in refugial centers, and subsequent expansion into postglacial habitats for hosts and parasites coincided with the cyclic pattern of stadials and interstadials. During the past 2-3 million years following colonization, these dramatic climatic fluctuations strongly influenced the continuity of ecological associations in marine habitats and appear to have been the determinants of congruent and synchronic patterns of speciation among these disparate taxa of marine homeotherms and eucestodes.

Sarcocystis falcatula of opossums: transmission by cockroaches with fatal pulmonary disease in psittacine birds.

Abstract: r: Old World psittacines experienced an acute fatal illness in outdoor breeding collections in South Florida. Toxoplasma-like organisms were found histologically in pulmonary capillaries and elsewhere. Because the organisms underwent schizogony and could not be transmitted to mice, we looked for a cause other than Toxoplasma gondii. An opossum was trapped on the premises of 1 facility and was found to be shedding sporocysts similar to Sarcocystis falcatula in its feces. Cockroaches were prevalent and suspected as transport hosts. Cockroaches that had ingested opossum feces and subsequently were fed to cockatoos induced an identical fatal illness. Obstruction of pulmonary capillaries by developing schizonts and pulmonary edema were the most important pathologic findings. The epidemic was stopped by biological insect control employing flightless chickens to reduce cockroach populations and by an electric fence restricting access of opossums to these outdoor psittacine breeding facilities. An acute fatal illness was observed during a 4-yr period in a breeding colony (A) of birds of the order Psittaformes in Florida. Although the collection consisted principally of American spe- cies, deaths occurred mainly in non-American psittacines. An intensive search for the identity and source of the organism was undertaken. An- other outbreak occurring at psittacine facility B, approximately 22 km away, also was studied. Deaths occurred sporadically over the year.

Peptidergic messengers : a new perspective of the nervous system of parasitic platyhelminths

Abstract: Platyhelminths represent a strategic phylogenetic stage in the evolution of the nervous system. They are the most primitive animals alive today exhibiting bilateral symmetry with attendent cephalization and condensation of neurons into a central nervous system. Not surprisingly, their free-living members, the turbellarians, have been the subject of numerous behavioral, physiological, and anatomical studies from which sufficient information has been gleaned to indicate that much of the basic neuronal machinary of higher organisms is present in flatworms (Koopowitz, 1989; Gustafsson, 1990). Thus, even in lower organisms the so-called "simple" nervous system appears to be singularly complex, if less elaborate than that of higher organisms. But what of the nervous system in parasitic platyhelminths? Has parasitism led to a reduction or secondary simplification of the flatworm nervous system? Historically, the nervous system of parasitic platyhelminths has been difficult to evaluate. For example, it never has been an easy system to delineate or identify anatomically, using conventional staining with methylene blue or Golgi stains or with gold and silver chloride impregnations, nor does it lend itself readily to ultrastructural reconstruction by serial sectioning. In common with other organ systems in acoelomates, the flatworm nervous system is embedded in parenchymatous tissue and is therefore virtually impossible to dissect free from the rest of the body; and the small size of most parasitic organisms and the physiological problems of maintaining them in vitro present enormous technical difficulties for even the most deter-

Helminth parasites of the raccoon (Procyon lotor) from north-central Arkansas.

Abstract: Twenty-three species of helminths (4 trematodes, 2 cestodes, 14 nematodes, and 3 acanthocephalans) were found in the gastrointestinal tract, lungs, pancreas, tongue, urinary bladder, and subcutaneous tissues of 30 live-trapped or hunter-shot raccoons from north-central Arkansas between November 1989 and April 1990. Helminths were not detected in the brain, diaphragm, gallbladder, heart, liver, reproductive system, or trachea. Each raccoon examined was infected with 3 or more of the following helminths: Brachylaima virginiana, Eurytrema procyonis, Fibricola cratera, Pharyngostomoides procyonis, Atriotaenia procyonis, Mesocestoides spp., Arthrocephalus lotoris, Capillaria aerophila, Capillaria plica, Capillaria procyonis, Capillaria putorii, Crenosoma goblei, Cruzia americana, Dirofilaria tenuis, Dracunculus insignis, Enterobius sp., Gnathostoma procyonis, Molineus barbatus, Physaloptera rara, Trichinella spiralis, Centrorhynchus wardae, Macracanthorhynchus ingens, and Oligacanthorhynchus tortuosa. All helminths collected with the exception of D. insignis constitute new geographic distribution records. Occurrences of C. aerophila, C. wardae, and O. tortuosa are new host records. One nymphal pentastome, Porocephalus crotali, was found in the liver of 1 raccoon, constituting a new host record.

Infections of Gyrodactylus bullatarudis and Gyrodactylus turnbulli on guppies (Poecilia reticulata) in Trinidad.

Abstract: Gyrodactylus bullatarudis Turnbull, 1956, and Gyrodactylus turnbulli Harris, 1986, are recorded from guppies (Poecilia reticulata) from the northern mountains of Trinidad. Mixed infections of the 2 spe- cies were found at 9 localities. Gyrodactylus turnbulli had a predominantly posterior distribution on the fish- es, whereas G. bullatarudis was more anteriorly dis- tributed. This is the first record of these species from guppies collected from within their original range.

Biological tags for stock separation in Pacific herring Clupea harengus pallasi in California.

Abstract: A survey of the parasites of Pacific herring (Clupea harengus pallasi) off northern California identified 1 species of Acanthocephala, 1 species of Cestoda, 2 species of Copepoda, 1 species and 1 family of Digenea, 3 species of Nematoda, and 3 species of Protozoa. From this survey, Lacistorhynchus dollfusi (Cestoda), Parahemiurus merus (Digenea), and Anisakis simplex, Contracaecum sp., Hysterothylacium sp. (Nematoda) were selected as potential tags. Herring were collected in Tomales, San Francisco, and Monterey bays for the following 9 yr and examined for these select parasites. The results suggest that these parasites can be used to distinguish the spawning stocks of San Francisco and Tomales bays. The distribution of the definitive hosts of the respective parasites suggests that the Tomales Bay fish are offshore during the nonbreeding season and the San Francisco Bay fish onshore. The similarity in parasitism between San Francisco Bay and the nonspawning population in Monterey Bay suggests that these 2 populations represent a single stock.

Ribosomal DNA restriction analysis and synthetic oligonucleotide probing in the identification of genera of lower trypanosomatids.

Abstract: ABSTRATr: Fifty-four species or isolates of insect trypanosomatids were examined for the presence of selected restriction enzyme sites in the small (SSU) and large (LSU) rRNA coding units of ribosomal genes. In the SSU, sites for Eco RI, Bgl II, Pst I, and Hind III were found to occur at the same location for all species examined, thus displaying a universal distribution among trypanosomatids. In the LSU, a site for Bgl II in the 24S-alpha sequence and sites for Hind III and Pst I in the 24S-beta sequence were found in all species examined. In contrast, a site for Pvu II in the SSU exhibited a genus-related distribution, being present in Crithidia and Herpetomonas but absent in Phytomonas. A site for Hind III in the 24S-alpha sequence of the LSU also exhibited genus-restricted distribution. The site was present in Crithidia but absent in Phytomonas and Herpetomonas. These findings were confirmed by dot hybridization with a synthetic oligonucleotide complementary to the 18S rRNA sequence containing the Pvu II site. Results point to the usefulness of restriction markers as diagnostic tools for distinguishing the lower trypanosomatid genera Crithidia, Herpetomonas, and Phytomonas at the same time revealing a marked complexity within the genus Leptomonas. Proper identification of genera of trypanosomatid parasites of insects still is an unsolved