Search

Molecular characterization studies on Setaria equina are limited. The present study aimed to characterize S. equina at the cytochrome c oxidase gene and to examine its phylogenetic relationships with other filarid species. Sequence analysis showed 100% nucleotide homology with an S. equina sequence from Italy (AJ544873). However, both sequences exhibited 7 nucleotide substitutions from a S. equina donkey isolate from Egypt (MK541847). Overall, S. equina formed a monophyletic sister group to Setaria tundra. All Setaria spp. examined formed a separate group on the phylogenetic tree that was related to corresponding Onchocerca spp. and Dirofilaria spp. clades. Human filarid worms-Brugia spp. and Wuchereria spp. grouped in a separate clade alongside Theilezia spp. Dipetalonema spp.-formed a separate group at the top of the tree.

Although little studied, infections with nematodes of the Onchocercidae Leiper, 1911, predominated by the genera Dipetalonema Diesing, 1861 and Mansonella Faust, 1929, are frequent in wild primates and human populations in the Neotropical forest areas. This study reports natural infections with Dipetalonema freitasi Bain, Diagne & Muller, 1987 and D. gracile (Rudolphi, 1809) in two free-living species of pitheciid primates, extending the known geographical distribution of these species to the forest of the Peruvian Amazon. Adult worms were recovered from the thoracic and abdominal cavities of two species of monkeys, Pithecia monachus monachus (E. Geoffroy Saint-Hilaire) and Cacajao calvus ucayalii (Thomas) (Primates: Pitheciidae), collected along the Yavari-Mirin River basin and analysed via light and scanning electron microscopy. Both host species represent new host records for D. freitasi and D. gracile. Morphometric data are also presented for the sampled filarial worms in addition to morphological details obtained through light and electron microscopy examination of D. freitasi specimens.

Canine arthropod-borne infections are of major interest in small animal practice and have been widely investigated in Central and Western Europe. However, only limited epidemiological data are available from South-Eastern European countries, although diseases including babesiosis or dirofilariosis are widely recognised as important canine infections in these countries. A steadily increasing number of dogs imported from South-Eastern Europe into Germany require particular attention by small animal practitioners. In this study, a total of 216 dogs [29 local Romanian pet dogs presented at Salvavet Veterinary Clinic in Bucharest, Romania, and 187 imported stray dogs from Romania (n = 109) and Hungary (n = 78) into Germany] were screened by molecular biological, serological and haematological methods for canine arthropod-borne infections. Eleven different parasitic and bacterial vector-borne pathogens-Babesia canis canis, Babesia canis vogeli, Babesia gibsoni, Babesia felis-like, Hepatozoon canis, Leishmania spp., Dirofilaria immitis, Dirofilaria repens, Acanthocheilonema reconditum, Anaplasma phagocytophilum and Mycoplasma haemocanis-were detected. Fifty-six percent of the dogs were positive by direct methods. B. canis canis was the most prevalent pathogen in dogs imported to Germany (42.8%) and dogs submitted for clinical consultation in Bucharest (44.8%). Our data strongly suggest the introduction of an adjusted screening panel in dogs from South-East Europe in view of increasing importation of dogs into Germany.

The heartworm Acanthocheilonema spirocauda (Leidy, Proc Acad Nat Sci Philadelphia 10:110-112, 1858) Anderson, 1992 is described from material collected from harbour seals in Scandinavia and compared with types and other specimens described by Anderson (Can J Zool 37:481-493, 1959) from harbour seals in eastern USA. Most morphometric characters of the material from USA fall within the ranges established for the Scandinavian one. Some intraspecific variability in the organisation of papillae on the male tail was detected among the Scandinavian specimens. Differences between the specimens from Scandinavia and Eastern USA are also found in the organisation of papillae on the tail of males and females. An excretory pore was not discernible, but a clearly hemizonid-like structure is described. For the first time, scanning electron micrographs present external morphological structures of the species.

Mastomys coucha and jirds infected with Acanthocheilonema viteae, a filarial species free of endosymbiontic bacteria of the genus Wolbachia, suffer lethal side effects after effective microfilaricidal therapy with diethylcarbamazine and levamisole, whereas, M. coucha infected with the Wolbachia-infested species Brugia malayi or Litomosoides carinii tolerate corresponding treatment. Mortality in A. viteae infected, treated animals varied with microfilariae density in the blood. It was up to 100% in highly microfilaraemic M. coucha and jirds, but low or absent in animals with low microfilariae counts. Deaths occurred in most cases 5-24 h after treatment. Characteristic symptoms in animals, which died subsequently were a rapid drop in body temperature by 4-7 degrees C, an increase in hematokrit values by up to 10% and a moderate blood acidosis. Lethal effects in A. viteae infections did not depend on a particular status of hypersensitivity of the animals since desensitization procedures, which protected infected M. coucha against an otherwise lethal intravenous challenge with A. viteae homogenate did not protect against adverse reactions to a subsequent microfilaricidal treatment. The animals were protected from treatment induced death by injection of N-LMMA. Thus the final morbific agent seems NO. The data show that adverse effects after effective microfilaricidal therapy may be caused by microfilariae derived components different from Wolbachia-released LPS.

Four new species of Molinema (Filarioidea: Onchocercidae), parasites of echimyid rodents in South America, are described: M. algardneri n. sp. from Proechimys amphichoricus, M. barbarae n. sp. from P. cayennensis, both in Venezuela (Rio Negro and Cerro La Neblina, respectively), M. peruviensis n. sp. from P. steerei in Peru (lower Urubamba), and M. nattereri n. sp. (= Filaria diacantha Molin, 1858 pro parte) from Echimys ? didelphoides (= Loncheres rufa) in Brazil (Matto Grosso). They differ from each other and from the previously described species in the following characters: flat or concave head, cephalic ratio (distance between cephalic papillae in median view/lateral view), size and shape of the buccal capsule, length of the oesophagus, cuticular ornamentation of the female body, length of the ovijector, thick or slim female tail, area rugosa, long or short filament in the left spicule, heel in the right spicule, respective position of postcloacal papilla pairs 5 and 6, shape of the caudal lappets, terminal point present or absent, and the microfilariae. Molinema, which belongs to the worldwide Dipetalonema line, is distributed in two of the ten families of the Caviomorpha (South American Hystrichognathi have extended their distribution in South America since the early Oligocene). Nine species are parasitic in the Echimyidae (suborder Caviida); they have a short oesophagus and a complete or reduced set of precloacal papillae (four or three pairs). Two species are parasitic in the Erethizontidae (Erethizontida) and might be more primitive in having a well-developed oesophagus. However, the type-species M. diacantha, of which a female specimen was studied, has a large buccal capsule and has evolved concave head, while M. arbuta has a reduced buccal capsule and primitive flat head. This last species is a parasite of a Nearctic porcupine and probably represents a small line of Molinema which migrated to the north with its hosts when communications were established between the two American continents 3 mya (Pleistocene). M. sprenti, the unique parasite of the Castoridae in North America, is considered to be derived from this group by capture.

A purified 41-kDa protein of the rodent filaria Acanthocheilonema viteae was shown to protect jirds against a challenge infection. Subcutaneous immunization with the protein reduced the number of adult worms by up to 65% and the number of circulating microfilariae declined by up to 93% in these animals. The protein is located in the muscle tissues of adult worms and was identified as tropomyosin by N-terminal sequencing of the purified protein.

The effects of K+ on muscle contractility were explored in the filarial nematode Acanthocheilonema viteae (Dipetalonema viteae). The parasite was slit open longitudinally and mounted in a smooth muscle chamber that was filled with aerated (95% N2-5% CO2) physiological solution at 37 degrees C. KCl at concentrations ranging from 20 to 100 mM induced a rapid isotonic contraction of the filarial muscle. The maximal response from KCl was similar to the maximal response to acetylcholine chloride (ACh). When KCl was applied for several minutes, tolerance frequently occurred. Contractions were also induced by K2SO4 but not by NaCl, Na2SO4 or sucrose. Nifedipine was more than 10 times as potent in reducing the KCl-induced contraction as in reducing that caused by ACh. The KCl-induced contraction was abolished in a Ca-free physiological medium containing ethyleneglycol-bis-(beta-aminoethyl ether) N, N, N', N'-tetraacetic acid (EGTA, 10(-4) M). Low [Ca2+]/[Mg2+] solutions blocked the spontaneous activity, the KCl-induced contractions, and the ACh-induced contractions. KCl also induced contractions in denervated muscle strips, supporting the hypothesis that K+ acts directly on the muscle cells. These results indicate that K+ can depolarize the muscle membrane and induce a muscle contraction that is dependent on extracellular calcium ions.

Background Hippobosca longipennis (the 'dog louse fly') is a blood sucking ectoparasite found on wild carnivores such as cheetahs and lions and domesticated and feral dogs in Africa, the Middle East and Asia, including China. Known as an intermediate host for Acanthocheilonema dracunculoides and a transport host for Cheyletiella yasguri, it has also been suggested that H. longipennis may be a vector for other pathogens, including Acanthocheilonema sp.? nov., which was recently reported to infect up to 48% of dogs in northern India where this species of fly is known to commonly infest dogs. To test this hypothesis, hippoboscid flies feeding on dogs in Ladakh in northern India were collected and subjected to microscopic dissection. Results A total of 12 infective larvae were found in 10 out of 65 flies dissected; 9 from the head, 2 from the thorax and 1 from the abdomen. The larvae averaged 2, 900 (± 60) μm in length and 34 (± 5) μm in width and possessed morphological features characteristic of the family Onchocercidae. Genetic analysis and comparison of the 18S, ITS-2, 12S and cox-1 genes confirmed the identity of the larvae as the Acanthocheilonema sp.? nov. reported in dogs in Ladakh. Conclusion This study provides evidence for a potential intermediate host-parasite relationship between H. longipennis and the canine Acanthocheilonema sp.? nov. in northern India.

Increasing single doses of 5, 15, 30, 60 and 90 third-stage Acanthocheilonema viteae larvae per animal were inoculated into jirds. The adult worm load increased steadily, whereas the recovery rate decreased significantly, i.e. the correlation between dose and recovery was negative (rs, -0.90; n = 5; alpha, 0.05). The same inoculation doses were given as trickle inoculations of 5 L3 each (3 x 5, 6 x 5, 12 x 5, 18 x 5) at intervals of 2-6 days throughout the prepatency period. Irrespective of the number of repeated inoculations, a rather constant but low load of 7-10 worms/animal was reached. The recovery rate decreased drastically (rs, -1.0; n = 5). When trickle inoculations were carried out in animals exhibiting patient infections, the superinoculated larvae seemed to be destroyed almost completely; thus, a parasite-host equilibrium was guaranteed in all cases. The immunological background is discussed.

A first description of an adult filarial parasite of British and Irish greyhounds which produces microfilariae similar to those of Dipetalonema reconditum but which are shorter and lack the characteristic buttonhook tail. Adult worms recovered also resemble to some extent D. reconditum, but males have larger spicules and a different arrangement of the posterior papillae. Although these specimens have been tentatively assigned to the species D. reconditum, further studies are required to elucidate their exact taxonomic relationship.

The cuticle of the filaria Dipetalonema viteae was isolated by sonication in 1% sodiumdodecylsulphate (SDS) and in a mixture of 1% SDS and 5% B-mercaptoethanol (BME). Sonication in SDS removed all internal parts and left the cuticle intact; this was verified by light- and electronmicroscopy. Sonication and incubation of the cuticle in the mixture of SDS-BME at pH 6.8 dissolved the basal and part of the median zone of the cuticle. The epicuticle and the cortical zone remained intact. The extracts were investigated using SDS-polyacrylamide gel electrophoresis; the early extracts contained a wide variety of proteins, whereas the later steps showed a consistent pattern with a smaller number of bands. Cuticles after SDS-purification, the extract of cuticular material in SDS-BME, and the cuticles insoluble in SDS-BME were used to immunize mice; the antibodies produced were visualized by an indirect fluorescent antibody test on cryostat sections of female worms. When SDS-purified cuticles were used for immunization, antibodies directed against all organs in the filariae were found. The SDS-BME extract and the insoluble cuticular pellet stimulated the production of antibodies restricted to the cuticle of adult worms and microfilariae. The purification method opens up the possibility of further isolation and characterization of antigens from the cuticle.

Several groups of control and filaria-infected Rodents were treated by radio-labelled diethylcarbamazine in order to study the effect of parasitism on the protein binding and tissue distribution of the drug. These studies were performed using equilibrium dialysis, chromatography and autoradiography. The plasmatic protein concentrations were lower in the group of infected Rodents, due to a decrease of albumin. The binding rate depended on the electrophoretic fractions, but remained low and unaffected by parasitism. The radioactivity of DEC and its metabolites appeared very quickly in the tissues, particularly in infected Rodents. This was due to the fact that the already rapid intestinal absorption and diffusion were enhanced by filariasis.

Using a Litomosoides carinii adult antigen, time courses of antibody levels were followed by an ELISA in L. carinii, Dipetalonema viteae, Brugia malayi and B. pahangi infected Mastomys natalensis. Using various groups of infected animals, periods up to 400 days after infection were covered. In L. carinii infected Mastomys, antibodies were first detected 11 days p.i. and levels increased rapidly until day 40. Temporarily reduced levels about the beginning of patency were followed by increasing values until about 100 days p.i. Then the antibody content of the sera remained more or less constant until about 250 days p.i. although maximum levels were found at day 170. Thereafter, the antibody concentration in the sera declined slowly but high levels were still observed 390 days p.i. The antibody content was usually higher in animals with high microfilariae densities than in those with low microfilariae counts but relations could not be proven statistically. In D. viteae infected Mastomys, maximum antibody values were reached within the beginning of patency. Levels were not altered markedly until about 110 days p.i. Thereafter they decreased slightly but then remained constant until the end of the investigation period 350 days p.i. B. malayi infected animals showed a rapid increase of the antibody content in the sera; a maximum was reached by 20 days after the infection. Thereafter, somewhat constant levels were found for 4--5 months. After 300 days p.i. the antibody levels declined progressively, accompanied with increasing parasitaemia densities; after 380 days the levels reached about 2/3 of the maximum. However, despite this, no relation was found between the levels of parasitaemia and antibody in individual animals. In B. pahangi infections the main prepatent antibody increase occurred during week 5 p.i., when maximum values were observed. The beginning of patency and the early patency were accompanied with slightly declining antibody levels. From 150 days p.i. until the end of the investigation 400 days p.i., the antibody content of the sera was fairly constant.

Development of third-stage larvae of Dipetalonema viteae within subcutaneously implanted micropore chambers proceeded in all hosts tested up to the fourth-stage larvae and occasionally to adolescent worms. In the jird the timing of development was comparable to a natural infection. Although the mouse is an insusceptible host, larval development could take place, but was very slow. Two intraperitoneal inoculations of living third-stage larvae into mice induced the production of antibodies against the larval cuticle and against common antigens. In such immune mice the development of third- and fourth-stage larvae within micropore chambers was significantly inhibited, larval mortality was increased, and the larval motility was impaired.

In quantitatively infected Meriones unguiculatus autopsied during prepatency, the embryonic development of fertilized eggs in the uteri of female Dipetalonema viteae commenced 29-32 days p.i., and approximately 20 days later the first microfilariae were released. In host animals examined at the onset of patency 45-59 days p.i. the average adult recovery rate was 31%. The worms were localized in the subcutaneous tissue (65.3%), fasciae of muscles (26.9%) and body cavities (7.8%). Female worms from the peritoneal and pleural cavities were significantly longer (44.3 +/- 10.4 mm) than those from the other habitats (33.7 +/- 5.5 mm). Female worms 14-20 weeks p.i. contained a total of 172,700 +/- 62,400 intrauterine stages, 21% of them pathologically altered. Based on these parameters, the expected in vivo fecundity of a female worm was calculated as about 7000 microfilariae per day.

The metabolism of diethylcarbamazine (DEC) in healthy and filaria-infected Proechimys oris Rodents was investigated. DEC and14C labelled DEC were orally administrated (100 mg/kg). The drug was quickly and intensively metabolised. About 90% of the dose administered was eliminated in the urine within 24 hours. Monodeethylation was the major pathway. The major urinary metabolites were ethylcarbamazine and its N-oxide. Traces of N methylpiperazine 2,5 dione and their N-oxide were also found. Elimination was faster in filarial than in healthy animals, but metabolites were the same.

In cats infected with Brugia pahangi, antibodies first appeared against the larvae (L3), then against the adults (L5) and the microfilariae (mf). Homologous antigens were better than antigens prepared from heterologous species (Dirofilaria immitis, Dipetalonema viteae, Litomosoides carinii and Onchocerca gutturosa) in detecting antibodies to B. pahangi in the infected cats by indirect fluorescent antibody test (IFAT). Metabolic products of L5, but not L3 or mf, of B. pahangi were antigenic and were used in the enzyme-linked immunosorbent assay (ELISA) for detection of antibodies. Using various homologous antigens, IFAT was found to be more sensitive than counter immunoelectrophoresis and ELISA in the detection of antibodies in the infected cats. The best antigen was cryosections of L3, with a positivity rate of 81%. However, using L3, L5 and mf antigens in IFAT, a total positivity of 97% was obtained.

A method of detecting microfilariae ofDirofilaria andDipetalonema species by direct microscopic examination of the ‘buffy coat’ and plasma fractions of the microhaematocrit offered a ready means of screening blood samples for evidence of infestation.