Removal and identification of a subconjunctival parasite by light and scanning electron microscopy

Background: Ophthalmic dirofilariasis is an uncommon zoonotic parasitic infection caused by species of Dirofilaria, a dog tapeworm that is transmitted to human by mosquitoes. Man is a dead?end host for the parasite. Ophthalmic involvement is rare and includes periorbital, subconjunctival, subtenon, and intra?ocular involvement. We report the removal of a subconjunctival worm and identification by light microscopy (LM) and scanning electron microscopy (SEM). Purpose: A 62?year?old female presented with complaints of redness, discharge, and foreign body sensation with difficulty in opening eyes in the left eye for the last 3 days. The patient is a non?vegetarian. On examination, her best corrected visual acuity in both eyes was 20/20. On slit lamp examination, there was a long, thin, round, coiled white subconjunctival live worm in the left eye superiorly. The rest of anterior segment evaluation, intra?ocular pressure, and fundus was normal in both eyes. The parasite was removed under local anesthesia from subconjunctival space [Video]. External surface morphology under LM revealed fine transverse cuticular striations with tapered cephalic and caudal ends. Uterus was long and coiled with indistinguishable masses inside. The finding was also confirmed by SEM. Synopsis: A subconjuctival parasite was removed and identified as Dirofilaria repens by characteristic LM and SEM findings. Highlight: Dirofilaria species may lodge in many tissues of human bodies including eye and adnexa. Dirofilaria is a natural parasite of carnivorous animals, mostly dogs, cats, and foxes.[1] The most common mode of transmission to human is usually by bite of mosquitoes like Culex and Aedes, which are considered as vectors, and it is often thought that parasitemia is because of accidental conduction.[1] Simple surgical removal of the worm is curative. After removal, the worm should be visualized directly under LM. All the internal structures of the transparent worm could be seen and compared with those under SEM.

Finding Wolbachia in Filarial larvae and Culicidae Mosquitoes in Upper Egypt Governorate

Wolbachia is an obligatory intracellular endosymbiotic bacterium, present in over 20% of all insects altering insect reproductive capabilities and in a wide range of filarial worms which is essential for worm survival and reproduction. In Egypt, no available data were found about Wolbachia searching for it in either mosquitoes or filarial worms. Thus, we aimed to identify the possible concurrent presence of Wolbachia within different mosquitoes and filarial parasites, in Assiut Governorate, Egypt using multiplex PCR. Initially, 6 pools were detected positive for Wolbachia by single PCR. The simultaneous detection of Wolbachia and filarial parasites (Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens) by multiplex PCR was spotted in 5 out of 6 pools, with an overall estimated rate of infection (ERI) of 0.24%. Unexpectedly, the highest ERI (0.53%) was for Anopheles pharoensis with related Wolbachia and W. bancrofti, followed by Aedes (0.42%) and Culex (0.26%). We also observed that Wolbachia altered Culex spp. as a primary vector for W. bancrofti to be replaced by Anopheles sp. Wolbachia within filaria-infected mosquitoes in our locality gives a hope to use bacteria as a new control trend simultaneously targeting the vector and filarial parasites.

Subconjunctival dirofilariasis caused by Dirofilaria repens: A case report with short review.

Human dirofi lariasis is a rare infection caused by fi larial worms of genus Dirofi laria. Subconjunctival dirofi lariasis has been reported from Asia, Europe, Italy and Africa. In India majority cases are from Kerala, Karnataka and very few from Maharashtra. We report the rare case of subconjunctival dirofi lariasis caused by Dirofi laria repens from Maharashtra, western India.

Human ocular dirofilariasis due to Dirofilaria repens in Sri Lanka

OBJECTIVE@#To identify worms obtained from patients with eye lesions and to describe the demographic factors of patients with ocular dirofilariasis.@*METHODS@#A retrospective descriptive study was conducted in 31 worm samples from 30 patients referred by consultant ophthalmologists between 2006 and February 2014. Data on age, sex and site of the lesion were ascertained from the details given in the referral letters. Morphological identification of the worm was based on the maximum width, length and appearance of the cuticle. The sex of the worm was determined by the width, length and presence or absence of vulva opening. PCR was performed using Dirofilaria repens specific primers to confirm the species of worms which couldnot be identified morphologically.@*RESULTS@#Most of the patients belonged to the age group of 40-49 years (mean age = 42 years). Majority of them were females (70%). Subconjunctival lesions were the most frequent presentation, while the rest (n = 4) were found on eyelids. Female worms were extracted from 18 cases, and 11 had male worms. One individual had both male and female worms in a single nodule. Adults were the most commonly affected. This pattern was different from the previous studies in Sri Lanka where the most common age group affected was younger than 9 years old.@*CONCLUSIONS@#The present study showed a considerably high incidence of ocular dirofilariasis, stressing the importance of implementing preventive measures to reduce the transmission of this zoonotic filarial disease.

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

Xenomonitoring of Different Filarial Nematodes Using Single and Multiplex PCR in Mosquitoes from Assiut Governorate, Egypt

Wuchereria bancrofti, Dirofilaria immitis, and Dirofilaria repens are filarial nematodes transmitted by mosquitoes belonging to Culex, Aedes, and Anopheles genera. Screening by vector dissection is a tiresome technique. We aimed to screen filarial parasites in their vectors by single and multiplex PCR and evaluate the usefulness of multiplex PCR as a rapid xenomonitoring and simultaneous differentiation tool, in area where 3 filarial parasites are coexisting. Female mosquitoes were collected from 7 localities in Assiut Governorate, were microscopically identified and divided into pools according to their species and collection site. Detection of W. bancrofti, D. immitis, and D. repens using single PCR was reached followed by multiplex PCR. Usefulness of multiplex PCR was evaluated by testing mosquito pools to know which genera and species are used by filarial parasites as a vector. An overall estimated rate of infection (ERI) in mosquitoes was 0.6%; the highest was Culex spp. (0.47%). W. bancrofti, D. immitis, and D. repens could be simultaneously and differentially detected in infected vectors by using multiplex PCR. Out of 100 mosquito pools, 8 were positive for W. bancrofti (ERI of 0.33%) and 3 pools each were positive for D. immitis and D. repens (ERI 0.12%). The technique showed 100% sensitivity and 98% specificity. El-Nikhila, El-Matiaa villages, and Sahel Seleem district in Assiut Governorate, Egypt are still endemic foci for filarial parasites. Multiplex PCR offers a reliable procedure for molecular xenomonitoring of filariasis within their respective vectors in endemic areas. Therefore, it is recommended for evaluation of mosquito infection after lymphatic filariasis eradication programs.

A Rare Human Case of Dirofilaria repens Infection in the Subcutaneous Posterior Thorax with Molecular Identification

The emergence of Dirofilarial infections in Asia including Vietnam is a clinically significant threat to the community. We here report a rare case of subcutaneous Dirofilaria repens infection on the posterior thoracic wall in a young woman presenting a painful, itchy, and palpable nodule. The adult worm was identified by mitochondrial cox1 and nuclear ITS-2 sequence determination. The diagnosis was additionally confirmed by 16S rRNA sequencing of the endosymbiont Wolbachia pipientis commonly co-existing with D. repens. This is a rare case of subcutaneous human infection on the posterior thoracic region caused by D. repens.

Human ocular dirofilariasis due to Dirofilaria repens in Sri Lanka

Objective: To identify worms obtained from patients with eye lesions and to describe the demographic factors of patients with ocular dirofilariasis. Methods: A retrospective descriptive study was conducted in 31 worm samples from 30 patients referred by consultant ophthalmologists between 2006 and February 2014. Data on age, sex and site of the lesion were ascertained from the details given in the referral letters. Morphological identification of the worm was based on the maximum width, length and appearance of the cuticle. The sex of the worm was determined by the width, length and presence or absence of vulva opening. PCR was performed using Dirofilaria repens specific primers to confirm the species of worms which couldnot be identified morphologically. Results: Most of the patients belonged to the age group of 40-49 years (mean age = 42 years). Majority of them were females (70%). Subconjunctival lesions were the most frequent presentation, while the rest (n = 4) were found on eyelids. Female worms were extracted from 18 cases, and 11 had male worms. One individual had both male and female worms in a single nodule. Adults were the most commonly affected. This pattern was different from the previous studies in Sri Lanka where the most common age group affected was younger than 9 years old. Conclusions: The present study showed a considerably high incidence of ocular dirofilariasis, stressing the importance of implementing preventive measures to reduce the transmission of this zoonotic filarial disease.

A rare case of subconjunctival dirofilariasis by Dirofilaria repens in rural Gujarat.

Dirofi lariasis is a worldwide zoonotic fi lariasis with over 782 cases reported so far from diff erent parts of the world. Human dirofi lariasis, caused by Dirofi laria repens, have been reported to occur widely throughout Asia, Europe, and Africa. It has not been widely recognized in India, however; several cases have been reported in last few years. There is probably a focus of human infection with D. repens in Kerala. Herein, we present a review of human infections by D. repens, along with a case report of subconjunctival dirofi lariasis from rural part of Gujarat.

Oral dirofilariasis.

Filariasis affecting animals can rarely cause infections in human beings through the accidental bite of potential vectors. The resulting infection in man, known as zoonotic filariasis occur worldwide. Human dirofilariasis, the most common zoonotic filariasis, is caused by the filarial worm belonging to the genus Dirofilaria. Dirofilarial worms, which are recognized as pathogenic in man can cause nodular lesions in the lung, subcutaneous tissue, peritoneal cavity or eyes. Oral dirofilariasis is extremely rare and only a few cases have been documented. We report an interesting case of dirofilariasis due to Dirofilaria repens involving buccal mucosa in a patient who presented with a facial swelling. The clinical features, diagnostic issues and treatment aspects are discussed. This paper stresses the importance of considering dirofilariasis as differential diagnosis for subcutaneous swelling of the face, especially in areas where it is endemic.

Identificación morfológica y molecular de filarías caninas en una comuna semi-rural de la Región Metropolitana, Chile / Morphological and molecular identification of canine filariae in a semi-rural district of the Metropolitan Region in Chile

Introduction: To date, there has been no definitive confirmation of the presence of zoonotic dirofilariasis in dogs in Chile. Objectives: To study the presence of dirofilarias in blood samples from dogs collected in a semi-rural district near Santiago and to compare their frequency in dogs with and without dermatological manifestations. Methods: We examined 100 blood samples for dog filariae infections using microscopic methods (modified Knott technique). 50 dogs presented dermatological symptoms or signs compatible with filarial infections and 50 were asymptomatic. ITS-2 and 12s rDNA gene amplification by PCR and sequencing were performed in samples microscopically positive for microfilariae. Results. We observed microfilariae in 22 dogs (22%). Of these, 16/50 (32%) were symptomatic and 6/50 (12%) were asymptomatic (p = 0.02). Morphologically, the majority of micro-filariae were similar to Dirofilaria repens, although many had a bigger size than previously described. Nucleotide sequencing of the amplified genes showed no more than 95% homology with the D. repens sequences available for comparison. D. reconditum and D. dracunculoides infections were also identified. Conclusions: These features might indicate the presence of new species of Dirofilaria or a D. repens close related variant in Chile.

Introducción: A la fecha no hay datos concluyentes en Chile respecto a la presencia de dirofilariasis zoonótica en perros. Objetivos: Identificar la presencia de dirofilarias en sangre de perros de una comuna semi-rural cercana a Santiago y comparar su frecuencia en animales con y sin manifestaciones dermatológicas. Materialy Métodos. Se examinó un frotis sanguíneo de 100 perros en busca de microfilarias mediante observación microscópica (técnica de Knott modificada). Cincuenta perros presentaban síntomas o signos dermatológicos que se han asociado a esta parasitosis y 50 eran asintomáticos. Se amplificaron los genes ITS-2 y 12s ADNr de filarías en las muestras con microfilarias al frotis, secuenciando los fragmentos amplificados. Resultados: Se observaron microfilarias en 22 perros (22%), 16/50 (32%) sintomáticos y 6/50 (12%) asintomáticos (p = 0,02). Morfológicamente, la mayoría de las microfilarias observadas fueron similares a D. repens; sin embargo, una gran proporción mostró un tamaño mayor al descrito para esta especie. Las secuencias nucleotídicas de los genes amplificados mostraron una homología no mayor al 95% con las secuencias de D. repens disponibles para comparación. Se identificaron además dos especies poco patógenas, D. reconditum por morfología y secuenciación genética y D. dracunculoides por morfología. Conclusiones: Los resultados indican la existencia de una nueva especie de Dirofilaria cercanamente relacionada a D. repens o de una variante de esta especie.

Dirofilaria repens in Vietnam: Detection of 10 Eye and Subcutaneous Tissue Infection Cases Identified by Morphology and Molecular Methods

From 2006 to 2010, hospitals in Hanoi treated 10 human patients for dirofilariasis. The worms were collected from parasitic places, and identification of the species was completed by morphology and molecular methods. Ten parasites were recovered either from the conjunctiva (n=9) or subcutaneous tissue (n=1). The parasites were 4.0-12.5 cm in length and 0.5-0.6 mm in width. Morphological observations suggested all parasites as Dirofilaria repens. Three of the 10 parasites (1 from subcutaneous tissue and 2 from eyes) were used for molecular confirmation of the species identification. A portion of the mitochondrial cox1 (461 bp) was amplified and sequenced. Nucleotide and amino acid homologies were 95% and 99-100%, respectively, when compared with D. repens (Italian origin, GenBank AJ271614; DQ358814). This is the first report of eye dirofilariasis and the second report of subcutaneous tissue dirofilariasis due to D. repens in Vietnam.

Dirofilariasis: A rare case report.

Human dirofilariasis is a zoonotic infection most commonly caused by Dirofilaria repens. It has not been widely recognized in India. There is probably a focus of human infection with D. repens in Kerala. We report the first case of dirofilariasis, from the Eastern-part of India, to the best of our knowledge. Among the documented cases of human dirofilariasis caused by D. repens, recorded in India, most of them had ocular infections and few had subcutaneous involvement of the face. This is the first case report of human dirofilariasis from India involving the lower part of human body.