PCR-coupled sequencing achieves specific diagnosis of onchocerciasis in a challenging clinical case, to underpin effective treatment and clinical management.

This study demonstrates the utility of a PCR-based DNA sequencing approach to make a specific diagnosis of onchocerciasis in a returned traveller. Although a clinical diagnosis was not possible, the surgical excision of a suprascapular nodule from this patient, combined with an histological examination of this nodule and PCR-based sequencing of DNA from a nematode from this lesion solved the case. The analysis of DNA sequence data confirmed the presence of Onchocerca volvulus infection, supporting an effective treatment-clinical management strategy for the patient.

Onchocerca lupi infection in Turkey: a unique case of a rare human parasite.

Onchocerpa lupi was first isolated from a wolf in Russia. Since then, canine ocular onchocercosis has been increasingly reported, particularly in Europe and the United States. It is thought that blackflies and midges are the vectors of transmission, and it is possible that these vectors could transmit the parasite to humans. The first human case of O. lupi in Turkey was reported in 2011. In this report we present the third human case of O. lupi infection in Turkey. Our patient was a 28-year-old male who displayed a painless, immobile mass under the conjunctiva. The mass measured 10 × 12 mm in size. Pathological examination of the surgically excised tissue was suggestive of infection by a filarial nematode. Subsequently, the parasite was identified as O. lupi through molecular analysis. All of the previously reported cases of O. lupi in both humans and dogs were more symptomatic than in our patient, Onchocerca infection should not be ruled out during the differential diagnosis of the subconjunctival and orbital cystic mass in instances where there is little to no inflammation. It is important to consider biopsy and carry out molecular analysis to identify the parasite.

Zoonotic Onchocerca lupi infection in a 22-month-old child in Arizona: first report in the United States and a review of the literature.

A 22-month-old girl presented with neck pain and stiffness and magnetic resonance imaging showed an extradural mass extending from C2 through the C4 level with moderate to severe compression of the cord. A left unilateral C2-C4 laminectomy was performed revealing an extradural rubbery tumor; a small biopsy was obtained. Examination of stained tissue revealed the presence of a parasitic worm that was identified as a gravid female Onchocerca lupi. A magnetic resonance imaging at 7 weeks follow-up showed a significantly decreased size of the enhancing lesion and the patient's symptoms gradually resolved. This is the first report of zoonotic O. lupi in the United States. The parasite has been reported in dogs and cats in the western United States, and from people in four cases reported from Europe. A great deal more needs to be learned, including full host range and geographic distribution, before we fully understand O. lupi infections in animals and man.

Redescription of Onchocerca lupi (Spirurida: Onchocercidae) with histopathological observations.

BACKGROUND: Onchocerca lupi is a dog parasite of increasing zoonotic concern, with new human cases diagnosed in Turkey, Tunisia, Iran, and the United States. Information about the morphology of this nematode is scant and a detailed re-description of this species is overdue. In addition, histopathological data of potential usefulness for the identification of O. lupi infections are provided. METHODS: Male and female nematodes, collected from the connective tissue of a dog, were examined using light microscopy and scanning electron microscopy (SEM), and an histological evaluation was performed on biopsy samples from periocular tissues. RESULTS: The morphological identification was confirmed by molecular amplification and partial sequencing of cytochrome oxidase subunit 1 gene. This study provides the first comprehensive morphological and morphometric description of O. lupi from a dog based on light microscopy, SEM, molecular characterization, and histological observations. CONCLUSIONS: Data herein presented contribute to a better understanding of this little known parasitic zoonosis, whose impact on human and animal health is still underestimated. The presence of granulomatous reactions only around the female adult suggests that the release of microfilariae from the uterus might be the cause of the inflammatory reaction observed.

Defining parasite biodiversity at high latitudes of North America: new host and geographic records for Onchocerca cervipedis (Nematoda: Onchocercidae) in moose and caribou.

BACKGROUND: Onchocerca cervipedis is a filarioid nematode of cervids reported from Central America to boreal regions of North America. It is found primarily in subcutaneous tissues of the legs, and is more commonly known as 'legworm'. Blackflies are intermediate hosts and transmit larvae to ungulates when they blood-feed. In this article we report the first records of O. cervipedis from high latitudes of North America and its occurrence in previously unrecognized host subspecies including the Yukon-Alaska moose (Alces americanus gigas) and the Grant's caribou (Rangifer tarandus granti). METHODS: We examined the subcutaneous connective tissues of the metacarpi and/or metatarsi of 34 moose and one caribou for parasitic lesions. Samples were collected from animals killed by subsistence hunters or animals found dead in the Northwest Territories (NT), Canada and Alaska (AK), USA from 2005 to 2012. Genomic DNA lysate was prepared from nematode fragments collected from two moose. The nd5 region of the mitochondrial DNA was amplified by PCR and sequenced. RESULTS: Subcutaneous nodules were found in 12 moose from the NT and AK, and one caribou from AK. Nematodes dissected from the lesions were identified as Onchocerca cervipedis based on morphology of female and male specimens. Histopathological findings in moose included cavitating lesions with multifocal granulomatous cellulitis containing intralesional microfilariae and adults, often necrotic and partially mineralized. Lesions in the caribou included periosteitis with chronic cellulitis, eosinophilic and lymphoplasmacytic infiltrate, and abundant granulation associated with intralesional adult nematodes and larvae. Sequences of the nd5 region (471bp), the first generated for this species, were deposited with Genbank (JN580791 and JN580792). Representative voucher specimens were deposited in the archives of the United States National Parasite Collection. CONCLUSIONS: The geographic range of O. cervipedis is broader than previously thought, and extends into subarctic regions of western North America, at least to latitude 66°N. The host range is now recognized to include two additional subspecies: the Yukon-Alaska moose and Grant's caribou. Accelerated climate change at high latitudes may affect vector dynamics, and consequently the abundance and distribution of O. cervipedis in moose and caribou. Disease outbreaks and mortality events associated with climatic perturbations have been reported for other filarioids, such as Setaria tundra in Fennoscandia, and may become an emerging issue for O. cervipedis in subarctic North America.

First description of nodular onchocercosis (Onchocerca jakutensis) in free-ranging Italian red deer (Cervus elaphus).

Onchocercosis is a vector-transmitted parasitic disease involving wild and domestic ungulates, humans, and dogs. Red deer (Cervus elaphus) host numerous Onchocerca spp. which have precise anatomic sites in the host and two species, Onchocerca flexuosa Wedl, 1856 and Onchocerca jakutensis Guba-now, 1964, are found inside subcutaneous nodules. Between September and November 2007, subcutaneous nodules were observed on both thighs in shot red deer of a Tuscany population. We observed cystic structures, surrounded by a fibrous capsule, containing nematodes. Filamentous worms were male and female; microfilariae were also described. Although morphologically we could not distinguish between O. flexuosa and O. jakutensis, genetic studies implicated O. jakutensis. This is the first report of this parasite in Italy.

A case of canine ocular onchocercosis in Portugal.

Onchocercosis is a newly recognized disease in dogs that has been reported with higher frequency in Europe and in the United States. We report a case of a 3-year-old male mongrel stray dog from the Algarve region (South Portugal) who had a retrobulbar granuloma containing a filaroid nematode of the genus Onchocerca. A gravid adult female parasite was embedded in a granulomatous inflammation adjacent to the sclera beyond the retina. The parasite was 191 to 267 mum in diameter (mean = 225 mum), surrounded by a cuticule and owing a uterus that was filled with small unsheated microfilariae. The cuticule consisted of two separated layers in longitudinal sections. The external layer had cuticular ridges and the internal layer contained striations. Sequencing of the COI and ND5 mitochondrial genes confirmed the identity of this parasite as Onchocerca lupi. Furthermore, the first sequence of the 12S mitochondrial gene is reported in this study.

Subconjunctival zoonotic onchocerciasis in man: aberrant infection with Onchocerca lupi?

In the past few decades, 10 cases of cryptic, zoonotic onchocerciasis, including two subconjunctival infections, have been reported in man. In the majority of cases, Onchocerca cervicalis, O. gutturosa or O. dewittei, which normally infect horses, cattle and wild boar, respectively, were responsible for the lesions. However, the taxonomic status of the parasites involved in the two subconjunctival infections, both of which were European, has never been unambiguously determined. In such infections, the acute phase appears to be characterized by conjunctivitis. A single, strongly coiled, immature, female worm was found incorporated in a large granulomatous nodule, in the ocular and peri-ocular tissues, in the chronic stage of each of the two eye infections. Several, patent, sporadic cases of subconjunctival O. lupi infection have recently been reported in dogs. In terms of the location of the worms, clinical signs and histopathology, these canine infections were very similar to those seen in the two human patients with eye infection. When the parasites recovered from human eyes were compared morphologically with the Onchocerca spp. infecting animals in Europe, they appeared to be most similar to O. lupi. Although O. lupi is normally a parasite of dogs, it may thus also be responsible for aberrant, zoonotic, subconjunctival infections in man.

Second case of zoonotic Onchocerca infection in a resident of Oita in Japan.

A non-gravid female Onchocerca was found in histopathological sections of a biopsy specimen taken from a painful nodule in the wrist of a 57-year old woman in Oita, in southern Japan. Six species of Onchocerca have been found in animals in Japan: two in wild bovids, one in equids, and three in domestic bovids of which one, Onchocerca sp., is only known by the microfilaria and infective stage. Distinctive morphological features of the worm, including a three-layered thick cuticle with prominent annular ridges at wide intervals, high somatic muscles and narrow lateral chords, resembled those of O. gutturosa, one of the three bovine Onchocerca species transmitted in the Oita region. However Onchocerca sp., which is also transmitted in this region, cannot be excluded. An ELISA test of the patient serum suggests that infections by Onchocerca spp. might be distinguished from those by Dirofilaria immitis, of which the number of human cases is increasing in Japan.

The identification of some Onchocerca spp of cattle by isoenzyme analysis.

The isoenzyme patterns of LDH, MDH, PEP, PGM, MPI, GPI and GAPDH were compared for 3 species of cattle parasites Onchocerca gutturosa, O. gibsoni and O. lienalis from England, Australia and Mexico. LDH and GAPDH had identical patterns in each species but the other enzymes clearly distinguished between the species. PGM was the same in O. lienalis and O. gibsoni, but separated them from O. gutturosa. MPI and GPI had similar isoenzymes in O. gutturosa and O. gibsoni, but distinguished them from O. lienalis. Different isoenzyme patterns for all 3 species were obtained with MDH and PEP. The potential use of these techniques towards a better understanding of the epidemiology of human onchocerciasis is discussed.