David Bruce Conn

Human waterborne parasites in zebra mussels ( Dreissena polymorpha) from the Shannon River drainage area, Ireland.

Zebra mussels (Dreissena polymorpha) from throughout the Shannon River drainage area in Ireland were tested for the anthropozoonotic waterborne parasites Cryptosporidium parvum, Giardia lamblia, Encephalitozoon intestinalis, E. hellem, and Enterocytozoon bieneusi, by the multiplexed combined direct immunofluorescent antibody and fluorescent in situ hybridization method, and PCR. Parasite transmission stages were found at 75% of sites, with the highest mean concentration of 16, nine, and eight C. parvum oocysts, G. lamblia cysts, and Encephalitozoon intestinalis spores/mussel, respectively. On average eight Enterocytozoon bieneusi spores/mussel were recovered at any selected site. Approximately 80% of all parasites were viable and thus capable of initiating human infection. The Shannon River is polluted with serious emerging human waterborne pathogens including C. parvum, against which no therapy exists. Zebra mussels can recover and concentrate environmentally derived pathogens and can be used for the sanitary assessment of water quality.

Accumulation of human waterborne parasites by zebra mussels (Dreissena polymorpha) and Asian freshwater clams (Corbicula fluminea)

Abstract. Zebra mussels (Dreissena polymorpha) and Asian freshwater clams (Corbicula fluminea) are nonindigenous invasive bivalves present in North American fresh waters that are frequently contaminated with human enteric parasites, Cryptosporidium parvum and Giardia lamblia. Six-week laboratory exposure of D. polymorpha and Corbicula fluminea to both parasites seeded daily at concentrations reported from surface waters demonstrated efficient removal of Cryptosporidium parvum oocysts and G. lamblia cysts by both bivalve species. The number of parasites in mollusk tissue progressively increased in relation to the concentration of waterborne contamination, and decreased after cessation of the contamination. Oocysts outnumbered cysts in the tissue of both bivalves, and more parasites were identified in D. polymorpha than in Corbicula fluminea; overall 35.0% and 16.3% of the parasites seeded, respectively. Because C. fluminea and D. polymorpha can accumulate human waterborne parasites in proportion to ambient concentrations, these species of bivalves can be effective bioindicators of contamination of freshwater habitats with Cryptosporidium and Giardia.

Human dirofilariasis due to Dirofilaria repens in Ukraine, an emergent zoonosis: epidemiological report of 1465 cases

The filarial nematode Dirofilaria repens is currently considered to be one of the most extensively spreading human and animal parasites in Europe. In Ukraine, reporting cases of dirofilariasis has been mandatory since 1975, and the disease was included in the national surveillance system for notifiable diseases. Up until December 31st 2012, a total of 1533 cases have been registered, with 1465 cases occurring within the previous 16 years. Most of the cases of dirofilariasis were registered in 6 regions: Kyiv, and the Donetsk, Zaporizhzhya, Dnipropetrovsk, Kherson and Chernihiv oblasts. In the years 1997–2002 the highest incidence rate was noted in the Kherson oblast in the south of the country (9.79 per 100 000 people), and the lowest in western Ukraine (0.07–1.68 per 100 000 people). D. repens infections were registered in all oblasts. Parasitic lesions were most often located in the head, the subconjunctival tissue and around the eyes. D. repens lesions were also found in the limbs, torso, male sexual organs, and female mammary glands. Dirofilariasis was diagnosed in persons aged from 11 months to 90 years old, most often among people between 21–40 years of age. Most patients had only one parasitic skin lesion; the majority of isolated nematodes were female. The results of our analysis point to a constant increase in D. repens dirofilariasis incidence in humans in Ukraine. Despite educational efforts, infections have become more frequent and the territory in which the disease occurs has enlarged to encompass the whole of Ukraine. Nevertheless, the Ukrainian sanitary-epidemiological services managed to achieve some measure of success, e.g. by creating a registration system for D. repens infections and establishing proper diagnostics for the disease.

The pathogenicity, localization, and cyst structure of echinostomatid metacercariae (Trematoda) infecting the kidneys of the frogs Rana clamitans and Rana pipiens.

Light and scanning electron microscopy were used to examine the localization and pathogenicity of echinostomatid metacercariae infecting the kidneys of leopard frogs, Rana pipiens, and green frogs, Rana clamitans. Cysts occurred predominantly in the ventrolateral renal cortex, and at least some were confined to the lumen of the Bowmans capsules. Each vermiform metacercarial body was enclosed by a spherical cyst wall that had a uniform thickness. The wall was composed of a homogeneous material containing basic and keratinlike proteins, with sulfated acid mucopolysaccharides on the outer surface. Most cysts were enclosed by a fibrous capsule of host origin, or were surrounded by an inflammatory focus. Fibrosis was always focal, but its degree varied between individual hosts and between different cysts within the same host. Some heavily encapsulated cysts were darkened and contained disintegrating worms. In heavily infected kidneys, confluence of fibrotic or inflammatory foci resulted in the displacement of functional renal tissue. These data suggest that infection by echinostomatids may impair renal function and that the hosts response affects parasite viability.

MESOCESTOIDES LINEATUS (GOEZE, 1782) (MESOCESTOIDIDAE): NEW DATA ON SPERM ULTRASTRUCTURE

Spermiogenesis and the ultrastructural characters of the spermatozoon of Mesocestoides lineatus are described by means of transmission electron microscopy, including cytochemical analysis for glycogen. Materials were obtained from a golden hamster (Mesocricetus auratus) after experimental infection with tetrathyridia metacestodes obtained from naturally infected lizards (Anolis carolinensis) from Louisiana. Spermiogenesis in M. lineatus is characterized by the orthogonal growth of a free flagellum, a flagellar rotation, and a proximodistal fusion. The zone of differentiation contains 2 centrioles associated with striated rootlets and a reduced intercentriolar body. The mature spermatozoon of M. lineatus lacks a mitochondrion, and it is characterized by the presence of (1) a single, spiraled, crested body 150 nm thick; (2) a single axoneme of the 9+‘1’ pattern of trepaxonematan Platyhelminthes; (3) a parallel and reduced row of submembranous cortical microtubules; (4) a spiraled cordon of glycogen granules; and (5) a spiraled nucleus encircling the axoneme.

The role of cellular parenchyma and extracellular matrix in the histogenesis of the paruterine organ of Mesocestoides lineatus (Platyhelminthes: Cestoda)

The cellular parenchyma of adult Mesocestoides lineatus consists primarily of muscle cells divided into myofibrils, myocytons with numerous greatly dilated cisternae of granular endoplasmic reticulum, and glycogen‐rich myocytons with lipid droplets and membranous whorl inclusions. The latter are connected to each other by numerous gap junctions. Other mesenchymally arranged cells in the parenchyma are calcareous corpuscle cells. The extracellular matrix (ECM) consists of an electron‐lucent ground substance and numerous filaments. During histogenesis of the paruterine organ wall, the myocytons surrounding the posterior end of the uterus undergo extensive flattening, followed by cellular deterioration. There is a concomitant reduction in ECM/cell ratio. The paruterine organ lumen dilates and then its uterine epithelium breaks down as the wall thickens, so that the late gravid paruterine organ consists of a thick wall of membranous sheets enclosing the egg mass. Parenchymal cells in other regions of the proglottid do not undergo change. These data provide evidence for epithelial‐mesenchymal‐ECM interactions in the development of an epithelio‐mesenchymal organ in the Platyhelminthes.

FINE STRUCTURE OF THE GRAVID PARUTERINE ORGAN AND EMBRYONIC ENVELOPES OF MESOCESTOIDES LINEA TUS (CESTODA)

Mesocestoides lineatus tetrathyridia were removed from the body cavity of lizards, Anolis caroli- nensis, and fed to hamsters in which they developed to adults. Gravid proglottids were taken from feces for study. Intact proglottids or excised paruterine organs (PO) were processed by standard techniques for TEM. For SEM, PO contents were exposed by subjecting whole proglottids to freon cryofracture or sectioning in Paraplast. Gravid POs were thick-walled with enclosed eggs distributed throughout a cellular matrix. The wall was of parenchymal origin, containing muscle bundles and various cellular products. Each egg consisted of an oncosphere enclosed by a double-unit oncospheral membrane and a syncytial cytoplasmic envelope containing an embryo- phore. A discrete outer envelope and capsule were absent. Eggs were separated by PO matrix cells with prominent nuclei, mitochondria, intercellular junctions, and much extracellular material. These matrix cells may be related ontogenetically to individual eggs and thus homologous to embryonic envelope cells normally lying outside the embryophore of other cyclophyllids. Some differences in embryonic envelopes presumably were associated with containment of all eggs in the PO which may eliminate the need for individual protective shells. Other components of the PO matrix such as large reticular bodies are of unknown significance.

Ultrastructure of the gravid uterus of Hymenolepis diminuta (Platyhelminthes : Cestoda)

The fine structure of the uterus in gravid proglottids of Hymenolepis diminuta was examined by standard techniques for scanning and transmission electron microscopy. The uterus consisted of a syncytial uterine epithelium attached to the medullary parenchyma through a thin extracellular basal matrix. The epithelium contained prominent nuclei in the juxtalumenal cytoplasm. The cytoplasm was dominated by extensive granular endoplasmic reticulum, with dilated cisternae containing an electron-lucent material and widely scattered electron-dense spherical bodies. No Golgi body or other agranular endomembrane component was observed, but the epithelium contained numerous free ribosomes and a few mitochondria. The apical plasma membrane was folded into long microlamellae. Epithelial and epitheliomesenchymal folds and villi resulted in a compartmentalized uterine lumen, with each chamber containing 1 to several eggs. These data suggest a high level of synthetic activity within the uterine epithelium, but the chemical products and functional significance of this activity are not yet known.

Morphology and fine structure of the gravid uterus of three hymenolepidid tapeworm species (Platyhelminthes: Cestoda)

Summary The ultrastructure of the uterus in gravid proglottids of Hymenolepis citelli, Vampirolepis nana and Vampirolepis microstoma was examined using transmission and scanning electron microscopy. The cellular and subcellular structures were similar in all three species, consisting basically of a syncytial layer attached to a basal extracellular matrix. All nuclei were juxtaluminal and each contained a single large centrally located nucleolus and prominent masses of central and peripheral heterochromatin. The cytoplasm showed evidence suggesting a high level of protein synthesis and secretion. It consisted primarily of granular endoplasmic reticulum with moderately dilated cisternae; most cisternae were completely filled with an electron-lucent flocculent material, but others contained electrondense granules. Free ribosomes and mitochondria were also present. The apical plasma membrane and a small amount of enclosed cytoplasm were folded into long microlamellae that extended into the uterine lumen. Larg...

Life cycle and postembryonic development of Oochoristica anolis (Cyclophyllidea: Linstowiidae).

Gravid proglottids of Oochoristica anolis from naturally infected anole lizards, Anolis carolinensis, were placed in covered Petri dishes with laboratory-reared beetles, Tribolium confusum and Tenebrio molitor. After maintenance at 25 C, metacestodes developed in 29 of 61 T. confusum (48%), but in none of 5 T. molitor. Beetles contained from 1 to 22 metacestodes (means = 3.3), which were fully developed by day 40 postexposure. A primary lacuna was never observed, but the possibility of its presence could not be ruled out without histological study. No cercomer was formed and metacestodes retained larval hooks throughout development. Scolices were invaginated at removal from the hemocoel, but usually evaginated quickly in Ringers. On day 60 postexposure, metacestodes were fed by stomach tube to 5 anoles, 2 lacertid lizards (Podarcis muralis) and 2 mice. Worms developed only in anoles, 3 of which were infected upon examination. Oncospheral hooks were present in worms after 7 days development in the lizard; a median excretory pore was present at the posterior tip of all stages examined, including the terminal mature proglottid of a worm after 105 days in a lizard. Scolex growth rate was linear throughout metacestode and adult development, but growth rate in body length was diphasic, punctuated by change of hosts, associated with strobilization. Attempts to establish parenteral infections in anoles were unsuccessful. Present data constitute the most complete life history study thus far for any species of Oochoristica.