Morphological and Molecular Characteristics of the Gastro-Intestinal Nematode Parasite Ascaridia columbae Infecting the Domestic Pigeon Columba livia domestica in Saudi Arabia.

BACKGROUND: Parasitism is a complex problem that is often ignored in companion animals, including birds, unless it develops into a severe clinical disorder. The present study was, therefore, aimed to investigate the presence of the gastrointestinal nematode infecting the domestic pigeon and provide a complete morphological description and clarify its taxonomic position through phylogenetic analysis of the ITS1-5.8s-ITS2 rDNA gene region. MATERIALS AND METHODS: During the current study, a total of twenty-six domestic pigeons, Columba livia domestica, were collected and internal organs examined for helminth detection. Using light and scanning electron microscopy, the recovered parasite species are studied. In addition, the selected gene region was obtained and sequenced using appropriate primers that aid in the formation of the phylogenetic dendrogram for the recovered parasite species with others retrieved from GenBank. RESULTS: Morphological examination showed that this nematode parasite belongs to the Ascaridiidae family within the genus Ascaridia. The material was assigned to the previously described Ascaridia columbae by providing all the characteristic features as the presence of a mouth opening surrounded by three tri-lobed lips; each lip has two triangular teeth with a spoon-like structure, cephalic papillae and amphidal pores on lips surface, presence of lateral cuticular alae and pre-cloacal sucker, 10 pairs of caudal papillae, and two equal spicules in male worms. The morphological investigations of this species were supplemented by molecular analysis of ITS1-5.8s-ITS2 rDNA gene region. The data showed that the present A. coulmbae is deeply embedded in the Ascaridia genus with a 74-99% sequence similarity to other species in the Chromadorea class. Ascaridiidae appears as monophyly and represented as a sister group to Heterakidae. The ascaridiid species examined belong to the Ascaridia genus and displaced a close relationship with the previously described A. coulmbae (gb| KF147909.1, gb| AJ001509.1, gb| KC905082.1, gb| JQ995321.1, gb| JX624729.1) as putative sister taxa. CONCLUSION: The present study revealed that the species Ascaridia is the first account of this genus as an endoparasite from the domestic pigeon inhabiting Saudi Arabia. Therefore, the combination of morphological and molecular studies helps to identify this species correctly and identified as Ascaridia columbae.

Ultrastructure of Ascaridia galli (Schrank, 1788) (Nematoda: Ascaridida) from the endangered green peafowl Pavo muticus Linnaeus (Galliformes: Phasianidae).

Ascaridia galli (Schrank, 1788) is a common parasite of various galliform birds worldwide. Although A. galli has been extensively studied by many author, knowledge of the morphology of this species in detail is still insufficient. In the present paper, the detailed morphology of A. galli was further studied using light and scanning electron microscopy, based on specimens collected from the endangered green peafowl Pavo muticus Linnaeus (Galliformes: Phasianidae) in China. The results revealed some erroneous and previously unreported morphological features, including the lips lacking real denticles, the lateral alae beginning at some distance posterior to the base of the ventrolateral lips and the caudal papillae with 4 different morphotypes. The present morphological and morphometric data complement previous descriptions and enable us to recognize this species more precisely.

In vitro anthelmintic activity of root-tuber extract of Flemingia vestita, an indigenous plant in Shillong, India.

The in vitro activity of root-tuber-peel extract of Flemingia vestita, an indigenous plant consumed by the natives in Northeast India, was tested against helminth parasites. Live parasites (nematode: Ascaris suum from pigs, A. lumbricoides from humans, Ascaridia galli and Heterakis gallinarum from domestic fowl; cestode: Raillietina echinobothrida from domestic fowl; trematode: Paramphistomum sp. from cattle) were collected in 0.9 % physiological buffered saline (PBS) and maintained at 37 +/- 1 degrees C. In vitro treatment of the parasites with the crude extract (50 mg/ml) in PBS revealed complete immobilization of the trematode and cestode in about 43 and 20 min, respectively. However, the cuticle-covered nematodes did not show any change in physical activity and remained viable even after a long period of exposure to the extract. Exposure of R. echinobothrida to genistein (0.5 mg/ml), an active principle isolated from the root-tuber peel, caused spontaneous loss of movement (paralysis) in 4.5 h, which was slower than the time required for praziquantel, the reference flukicide and cestodicide. The treated parasites showed structural alteration in their tegumental architecture. This study suggests the vermifugal activity of this plant extract against trematodes and cestodes.

Scanning electron microscopy of Ascaridia galli (Schrank, 1788), Freeborn, 1923 and A. columbae (Linstow, 1903).

The morphology of the two ascaridoid nematodes Ascaridia galli and A. columbae was studied by scanning electron microscopy. The two nematodes were compared together and their specific characteristics were established, including lips, cephalic papillae, body cuticle, spicules and caudal papillae of the male.

Biosynthesis of dolichyl diphosphate N-acetylglucosamine intermediates in Ascaridia galli microsomes.

1. N-acetylglucosamine-1-phosphate transferase was demonstrated in the microsomal fraction of Ascaridia galli. 2. The transferase reaction depends on exogenous dolichyl phosphate as lipid acceptor and was found to be inhibited by tunicamycin. 3. The enzyme activity was optimal in the presence of sodium deoxycholate as detergent and Mg cations after 10 min of incubation. 4. The product of the transferase reaction--dolichyl diphosphate N-acetylglucosamine was converted into lipid-disaccharide-dolichyl diphosphate N,N'-diacetylchitobiose. 5. The maximum level of the conversion was achieved at 5 mM concentration of unlabelled UDP-N-acetylglucosamine, while this conversion was negligible at lower UDP-N-acetylglucosamine concentrations (0.1 and 0.5 mM).

Nippostrongylus brasiliensis and Ascaridia galli: characterization of peroxisomes.

A method is described for the isolation of peroxisomes from mitochondria-enriched fractions obtained from both species of nematodes. The distributions of these organelles are characterized after density gradient centrifugation in sucrose or Percoll by urate oxidase and catalase activities. The possession of peroxisomes may be part of an important defence mechanism in parasites.

Protofilament number in microtubules in cells of two parasitic nematodes.

The parasitic nematodes, Ascaridia galli and Trichostrongylus colubriformis, were prepared for electron microscopy with fixatives containing tannic acid, which allowed their microtubule protofilament number to be examined. In contrast to many mammalian tissues, the nematodes did not contain microtubules with 13 protofilaments. Ascaridia galli contained microtubules with 11 protofilaments in all tissues examined, including nerve, intestinal, pharyngeal, and hypodermal cells. Trichostrongylus colubriformis contained nerve cells, known as microtubule cells, with bundles of larger microtubules (approximately 30 nm in diameter) with 14 protofilaments. The microtubules in these cells did not appear to be continuous for the entire length of the axon. Other cells examined in T. colubriformis, including nerve, intestinal and pharyngeal cells, contained two distinct types of microtubules, one with 11 protofilaments and an approximate diameter of 25 nm, and one with 12 protofilaments and an approximate diameter of 27 nm. All cell types examined contained both types of microtubules.

Nippostrongylus brasiliensis and Ascaridia galli: mitochondrial respiration in free-living and parasitic stages.

Aerobic respiratory pathways have been delineated and respiratory efficiency has been assessed in mitochondria isolated from embryonated eggs, infective larvae, and adult Nippostrongylus brasiliensis and Ascaridia galli. Mitochondrial respiration in free-living stages of N. brasiliensis is mediated mainly by a mammalian-like antimycin A- and cyanide-sensitive pathway; specific respiratory activity is high and oxidative phosphorylation efficient. In mitochondria of adult N. brasiliensis, antimycin A- and cyanide-sensitive respiration is decreased relative to respiration though an alternative pathway, and specific respiratory activity and mitochondrial efficiency are lower. Respiration in mitochondria from embryonated eggs and tissues of adult A. galli is comparable, and apparently mediated by an antimycin A- and cyanide-insensitive alternative respiratory pathway; no evidence for the presence of a mammalian-like respiratory pathway in embryonated eggs of A. galli was found. The results of this study are compared to mitochondrial respiration in eggs, larvae, and adult body wall muscle of Ascaris suum.