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PubMed1992

Apparent lack of genetic variation within Pelecitus roemeri (Nematoda: Filarioidea) from three Australian species of macropodid marsupial.

An electrophoretic study of Pelecitus roemeri from Macropus robustus, M. giganteus and Wallabia bicolor revealed no genetic differences at 23 enzyme loci. The genetic data support the existing morphological evidence that P. roemeri from these three hosts represents a single species. The data show no genetic variation between nematodes from the same or different host species collected in northern and southern Australia. This result is discussed briefly in relation to Price's model of parasite speciation.

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PubMedDate not supplied

Isoenzymatic diagnosis of filariae: a method for separation of lactate dehydrogenase isoenzymes from Molinema dessetae (Nematoda: Filarioidea).

Lactate dehydrogenase (LDH) is highly active in filariae and could be a valuable tool for phyllogeny studies. Unfortunately, the isoenzymatic diagnosis of filariae is often difficult for LDH because of a poor mobility of the enzymes in starch gels which are the most commonly used in such studies. We propose here a method to separate filarial LDH isoenzymes using disc electrophoresis. The experiments were carried out on male and female Molinema dessetae in order to compare their respective isoenzymes. The study of several parameters such as buffer systems, percentage of bisacrylamide and progression time led to optimize the enzyme separation. LDH from male and female filariae were compared to mammal LDH-H4 and LDH-M4. Five and four LDH isoenzymes were found, respectively, in male and female worms. Relative concentration of each isoenzyme diverged between male and female worms. Mammal muscle LDH-M4 type moved between LDH2 and LDH3 from female worms, and between LDH1 and LDH2 from male worms. Mammal heart H4 type enzyme was very different in electrophoretic mobility. The ratio of each isoenzyme was determined by densitometry. The major isoenzymes from female worms will be studied as a biochemical target for chemotherapeutic attack.

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PubMed1979

Dunnifilaria dilli sp. n. (Nematoda: Filarioidea) from Rattus koratensis in Thailand.

Dunnifilaria dilli, a new species of filariid from Rattus koratensis from Thailand is described. Adult worms were recovered from the lymphatic system. They differ from D. ramachandrani in their location in the host, the tail length, the shape of the spicules, and in the number of caudal papillae. Microfilariae are much smaller and unsheathed. Additional hosts include R. rattus and R. sabanus.

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PubMed1976

Studies on the Malayan forest rat filaria, Breinlia booliati (Filarioidea: Onchocercidae): course of development in rat host.

The development of Breinlia booliati is described in its natural host, Rattus sabanus and in an inbred strain of laboratory albino rat. The growth of the parasite is similar in both the rat hosts. The third moult occurs between six-eight days and the final moult between 24-28 days. Larvae were recovered initially from the skin and carcass. After five weeks, developing stages were seen only in the thoracic and abdominal cavities, the site of development of the adult worms. Worms became sexually mature by 11-12 weeks and there was considerable growth in length of the female worms after this stage.

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PubMed1982

The host-parasite relationships in normal and protein-malnourished cotton rats infected with Litomosoides carinii (Nematoda: Filarioidea).

A total of 120 young cotton rats were fed ad libitum for 10 weeks on either synthetic diets containing 2.5, 5, 10 or 15% casein or on a stock breeding diet containing 20% crude protein. Half of the animals in each group were infected with the filarial nematode Litomosoides carinii and the food intake and body weight of each animal was measured. There was a trend towards increased food consumption in protein-deficient cotton rats and this was increased further in infected animals; however, differences between groups were not significant. Low protein diets reduced the rate of growth of uninfected and infected cotton rats; filarial infection intensified these adverse effects of under-nutrition and increased the number of rats which died in the lowest protein group. Protein deficiency had little effect on the number of L. carinii which developed, although fewer parasites became established in cotton rats fed on a 2.5% diet than in those fed on a 10% diet. Parasites developing in protein-deficient animals were shorter than those developing in well-fed ones and embryogenesis was retarded in female worms from protein-deficient animals. The onset of patency was retarded in protein-deficient animals and the microfilaraemia which developed subsequently was lower up to 120 days post-infection.

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PubMed1989

Neurosecretory-like material in 3rd- and 4th-stage Dirofilaria immitis larvae (Nematoda: Filarioidea).

Phase-interference microscopic examination of the infective, post-infective 3rd-stage and 4th-stage larvae of Dirofilaria immitis has identified a single cell body in each of the paired lateral amphidial nerves which undergoes characteristic morphological change during the development from 3rd- to 4th-stage larvae. Acetaldehyde-fuchsin staining of worm sections revealed fuchsinophilic material in the precise location of the amphidial nerve-cell bodies observed by phase-interference microscopy. This material was found in 70% of infective larvae recovered from mosquitoes and in 100% of larvae recovered from micropore chambers 24 h after implantation into BALB/C mice. In pre-moult larvae recovered at 42 h (Experiment 1) and at 48 h (Experiment 2) fuchsinophilic material was demonstrable, but no staining was observed in those larvae in which separation of the 3rd- and 4th-stage cuticles had occurred. No such material was observed in 4th-stage larvae recovered from chambers after 74 h, and in these larvae the amphidial nerve-cell bodies were not discernible. These cytological observations are consistent with a cycle of elaboration and release of neurosecretion associated with moulting in D. immitis.

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PubMed1986

Gametogenesis and fertilization in Dirofilaria immitis (Nematoda: Filarioidea).

Gametogenesis in Dirofilaria immitis has been studied principally by means of the aceto-orcein chromosomal squash technique, but with additional ultrastructural observations. A terminal germinative zone, in which a continuous and rapid division of germ cells occurs, has been identified in the gonoduct of both male and female worms. Approximately 20% of cells within these germinative zones were in arrested mitotic division following the incubation in vitro of excised gonads in 0.01% colchicine for 4 h. All primary spermatocytes within a 1-2 cm length of the testis proximal to the germinative zone were at the prophase of the 1st meiotic division. In the corresponding region of the ovary, the primary oocytes were similarly at the prophase of the 1st meiotic division in 75% of female worms examined but in the remaining 25% all primary oocytes possessed markedly less condensed, probably interphase nuclei. A possible hormonal control of the cyclical development of primary oocytes, but not primary spermatocytes in D. immitis is suggested. In most of the remaining length of the gonoducts beyond this region of cells at meiotic prophase, the chromatin material of both primary spermatocytes and oocytes is decondensed. Recondensation of chromosomes in the spermatocytes is observed just prior to entry into the seminal vesicle, where meiosis I is completed and meiosis II takes place. In the primary oocyte, completion of meiosis only occurs after fertilization within the seminal receptacle by an entire male gamete. Following the 2 meiotic divisions in the oocyte and subsequent extrusion of the 2 polar bodies, the haploid chromosome complement of the female unites with that of the male, re-establishing the diploid number of the zygote (2n = 10). Male chromosomes within the oocyte remain visible throughout late oogenesis and fusion occurs without the formation of pronuclei.

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PubMed1993

Purification and properties of phosphoenolpyruvate carboxylase from Molinema dessetae (Nematoda: Filarioidea).

The presence of phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31), an enzyme at the branchpoint of glycolysis and the Krebs cycle was detected in the Filaria Molinema dessetae. This enzyme has not previously been identified in Helminths, which have so far been found to only possess a phosphoenolpyruvate carboxykinase (EC 4.1.1.32). This enzyme had a level of activity comparable to that of pyruvate kinase, and was relatively less active than enzymes such as malate dehydrogenase or lactate dehydrogenase. We propose here a method of purification of M. dessetae PEP-carboxylase. When purified to electrophoretic homogeneity, the enzyme had a molecular weight of 64 kDa. Kinetic studies indicated that the carboxylation reaction had an optimal pH of 5.8. The enzyme was inhibited by cations such as Fe2+, Zn2+, Cd2+, Cu2+ but required the presence of Mg2+ or Mn2+. The enzyme was thermostable. The apparent Km value of 2.38 mmol for phosphoenolpyruvate for the carboxylation reaction was higher than previously reported values. The Km value for KHCO3 was found to be 1.6 mmol. PEP-carboxylase did not catalyse the reverse reaction.

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