Microscopic and molecular detection of Deraiophoronema evansi (Lewis, 1882) in domestic Bactrian camels (Camelus bactrianus) of Mongolia.

Cameline filarosis is an important parasitic disease having an economic impact on the camel industry around the world. However, there has been no study on filarosis in Bactrian camels of Mongolia. Therefore, the aim of the present study was to detect and identify microfilariae of Deraiophoronema evansi (D. evansi) in Bactrian camels from three provinces, located in southern and southwestern Mongolia. Blood samples were obtained from 400 healthy two-humped camels of different ages and both sexes. All blood samples were analysed using a variety of diagnostic techniques. Microfilariae were detected in 30 Bactrian camels (7.5%) by the Knott technique, while 13 Bactrian camels (3.3%) tested positive in a direct smear test. D. evansi was detected in 18 Bactrian camels (4.5%) by PCR assay. Prevalence was shown to be high among Bactrian camels in the age group up to 5 years, while the lowest positive results were obtained for Bactrian camels in the 5-10-year age group and the over 10-year age group. To confirm the morphological identification, D. evansi-COI gene sequences were subjected to phylogenetic analyses. The D. evansi-COI gene sequences from Mongolian two-humped camels were identical to sequences from Iranian one-humped camels and were clustered together with these sequences in the phylogeny. This is the first report of molecular detection and identification of microfilariae of D. evansi in Bactrian camels of Mongolia.

Multiple live subconjunctival dipetalonema: report of a case.

Parasitic infestations of the eye have been reported since centuries, affecting various parts of the eye. Some are subtle, coexisting with vision, while many others damage and destroy, in part or totally, the gift of sight. This report describes a patient with live subconjunctival dipetalonema infestation of the right eye, with 22 parasites removed live in one sitting from one eye.

A simple molecular method for discriminating common filarial nematodes of dogs (Canis familiaris).

Accurate diagnosis of canine filariosis is essential for choosing correct therapeutic approach. Therefore, reliable methods for discriminating among the different filarial infections in dogs are needed. The authors report simple and highly specific molecular methods that identify the three most common filarial nematodes of European dogs: Dirofilaria immitis, D. repens and Acanthocheilonema (syn. Dipetalonema) reconditum, based on (1) PCR amplifications of mitochondrial DNA (12S rDNA and coxI) with general filarial primers followed by digestion with restriction enzymes that generates band polymorphisms clearly discriminating the three species and (2) PCR amplifications with species-specific primers to support the restriction analysis, in particular in the case of multiple infections.

First record of Acanthocheilonema dracunculoides from domestic dogs in Namibia.

Acanthocheilonema dracunculoides was diagnosed in 2 dogs from Windhoek, Namibia, by acid phosphatase staining of microfilariae. This is the 1st record of A. dracunculoides in Namibia.

Specific polymerase chain reaction for differential diagnosis of Dirofilaria immitis and Dipetalonema reconditum using primers derived from internal transcribed spacer region 2 (ITS2).

Both Dirofilaria immiti and Dipetalonema reconditum may be found in blood of infected dogs but it is not easy to distinguish D. immitis from D. reconditum in morphology. We cloned and sequenced the contiguous internal transcribed spacer (ITS) region, ITS1-5.8S-ITS2, of these two different parasites and published on GenBank as AF217800 for D. immiti and AF217801 for D. reconditum in this study. We designed two pairs of specific primers derived from ITS2 being used for polymerase chain reaction (PCR). The amplicons of ITS2 from D. immiti and D. reconditum are 302 and 348bp, respectively. Moreover, the limitation for amplifying ITS2 gene using this PCR demonstrated that 1 x 10(-2) microfilaria of each species of parasite smashed or even with mixed samples could be detected and the PCR products were predicted as the same as that described above. Thus, D. immiti and D. reconditum could be differentially diagnosed by this specific PCR. Seventeen clinical cases were evaluated and all of them were correctly identified. In this study, ITS1-5.8S-ITS2 of D. immiti or D. reconditum were the first time sequenced and analyzed. No significant similarity of ITS1 and ITS2 between D. immiti and D. reconditum could be observed.

Histochemical differentiation of Dirofilaria immitis, Dirofilaria repens and Acanthocheilonema dracunculoides microfilariae by staining with a commercial kit, Leucognost-SP.

The diagnosis of canine heartworm infection is based upon the presence of circulating Dirofilaria immitis microfilariae or on techniques for the detection of serum antibodies or antigens. In the first of these, discrimination between D. immitis, D. repens and Acanthocheilonema dracunculoides microfilariae is based upon the acid phosphatase histochemical stain. In this paper, we propose an alternative technique for histochemical staining using a commercial kit test of naphthol-AS-OL (Leucognost-SP). This offers the advantages of speed and simplicity as compared to the standard Barka procedure.

1H magnetic resonance imaging and 31P magnetic resonance spectroscopy in experimental filariasis.

1H Magnetic resonance imaging and 31P magnetic resonance spectroscopy (MRS) have been carried out in experimental rodent filariasis, i.e., Acanthocheilonema viteae infection in the rodent host, Mastomys coucha. The T2-weighted image of the infected host shows fine hyperintense thread like structures of adult filariid nests in the cervical region. 31P MRS of normal and infected hosts, localized over the same region of interest, show seven major peaks corresponding to phosphomonoesters (including glucose-6-phosphate, fructose-6-phosphate, fructose-1-6-diphosphate, phosphorylcholine, and adenine monophosphate or AMP), inorganic phosphate, glycerophosphorylcholine, phosphoenolpyruvate, phosphocreatine and nucleoside di- and tri-phosphates. Concentrations of phosphomonoesters (PMEs) are higher in the normal rodent compared with the infected ones. In vivo 31P MRS provides a non-invasive assessment of tissue bioenergetics and phospholipid metabolism.

Exfoliative colpocytology: a method for the diagnosis of Capuchin monkey filariasis?

During vaginal fluid examinations (Papanicolaou) to study the physiological sexual cycle of Cebus sp., abundant Dipetalonema gracile microfilariae (110-160 microns x 4-5 microns, without a sheath) were encountered in the genital fluid, but not in peripheral blood. Considering the great difficulty in diagnosing this obscure parasitosis, exfoliative colpocytology was found to be an efficient diagnostic.

Specific serodiagnosis with adult Onchocerca volvulus antigen in an enzyme-linked immunosorbent assay.

The cross-reactivity of the blood from onchocerciasis, loiasis, and dipetalonemiasis was tested by a micro-ELISA technique, utilizing adult Onchocerca volvulus antigen and blood samples taken on filter paper. The average ELISA values (OD at 500 nm) were as follows: 0.58 in persons with O. volvulus microfilariae (n = 81), 0.49 in microfilariae-negatives from the same endemic area (n = 39), 0.15 in dipetalonemiasis (n = 27), and 0.25 in loiasis (n = 12), while those of 65 Dipetalonema perstans-negative people were markedly low (average 0.14) and that of 22 Loa loa-negatives, 0.22, respectively. This ELISA could successfully differentiate onchocerciasis from dipetalonemiasis and loiasis.

Evaluation of the diethylcarbamazine provocative test in the diagnosis of Wuchereria bancrofti infections in the Nigerian savanna and the effects on Dipetalonema perstans.

Large scale filariasis surveys in rural areas for microfilaraemia, especially of periodic types such as Wuchereria bancrofti are known to cause considerable administrative, technical and social problems. The present investigation was carried out in the population of two villages in the Malumfashi district of the Northern Nigerian savanna. From the survey results, the sensitivity and specificity of two techniques-day-time diethylcarbamazine (DEC) provocative test by blood smear and concentration, and night-blood examination by smear and concentration especially for W. bancrofti-were assessed. Day-time DEC provocative test proved to be efficient in terms of sensitivity and specificity, compared with the night-blood method, for W. bancrofti detection but less so for Dipetalonema perstans, the other blood microfilaria found in this population during these studies. A regression line between night-blood survey results for W. bancrofti and the results from day-time DEC provocative test was calculated. With the help of this regression line it is possible to estimate W. bancrofti microfilarial prevalence for night surveys, using the DEC provocative test results of day-time surveys. This can be done with minimal, but known, loss of accuracy and incurs fewer administrative, technical and social difficulties.

Dipetalonema from the eye of a man in Oregon, U.S.A. A case report.

A 21-mm filarial worm appeared suddenly in the anterior chamber of the right eye of a 32-year-old man in western Oregon. By a simultaneous irrigation-aspiration procedure, it was removed alive and only slightly damaged and was identified as a female Dipetalonema in the fourth stage of development. It was the third such case to be reported from western Oregon. In this and one other case the worms were morphologically similar to adult worms identified as Dipetalonema arbuta Highby 1943 from the body cavity of the porcupine (Erethizon dorsatum) and a similar species, Dipetalonema sprenti Anderson 1953, from the body cavity of the beaver (Castor canadensis).

Miniature anion-exchange/centrifugation technique for the diagnosis of microfilaraemia in the field.

The miniature anion-exchange/centrifugation (AEC) technique, developed originally for the detection of low parasitaemias in laboratory rodents, was adapted to field use for the diagnosis of trypanosomiasis in man in Africa and was tested in The Gambia. During this field study it was found that microfilariae of Dipetalonema perstans could also pass through the anion exchange column and appear in the centrifugate as 'medusa heads'. One locality-group (Mansafa Bolon) showed a generally higher prevalence and prevalences in women over 40 years old were higher than in the corresponding male groups in every locality. The potential usefulness of this technique in epidemiological studies of filariasis and ways of improving the accuracy of numerical estimates are briefly discussed.

The slide agglutination test for the diagnosis of filariasis in camels.

The slide agglutination test was adapted for the diagnosis of filariasis in camels, using an antigen prepared from the microfilariae by a simple lytic technique. The preliminary results were satisfactory as the test detected 86 per cent of the infected animals. Only 6 per cent of the healthy camels with no blood parasites or microfilariae in their blood gave positive results and no positive reactions were obtained from 18 animals suffering from Trypanosoma infection.

Studies on the diagnosis and treatment of human filariasis in Rhodesia.

Experiences in Rhodesia with various recovery techniques available for the laboratory diagnosis of infections with Dipetalonema perstans and Wuchereria bancrofti are discussed. A diagnostic laboratory regimen for routine filarial investigations is suggested. Included are preliminary observations on the use of mebendazole (Vermox) for the treatment of D. perstans infections.