First record of Dioctophyme renale (Goeze, 1782) in the pampas fox (Lycalopex gymnocercus, Fischer, 1814) (Carnivora: Canidae).

Dioctophyme renale (Goeze 1782) has not previously been reported in the pampas fox (Lycalopex gymnocercus) (Fisher 1814), the most abundant canid of southern South America. A wild adult pampas fox female was found dead due to unknown causes in Santa Fe province, Argentina. Post-mortem examination revealed three red worms measuring 10, 11 and 15 cm long, each with an approximate width of 5 mm. All of them were found free in the abdominal cavity. The worms were all male and were identified through morphological examination and molecular analysis as D. renale. No worm was found in the kidneys. This study reports the first case of dioctophymatosis in the pampas fox in Argentina, increasing the range of wild aberrant host species infected by the giant kidney worm in the Neotropical region.

High prevalence and concomitant infection of Ranavirus and Eustrongylides sp. in the invasive American Bullfrog in Brazil.

American Bullfrogs, Aquarana catesbeiana, are invasive anuran species distributed worldwide. One of the adverse impacts that this species causes in native communities is as a reservoir host for pathogens and parasites. Here, we report the coinfection of two pathogenic organisms in A. catesbeiana: Ranavirus and the nematode Eustrongylides. Bullfrogs were collected in the wild in a pond close to the urban area of São Paulo, Brazil. The prevalence of both pathogens was high: 77% were infected with ranavirus with a mean viral load of 1010.3 viral copies, and 100% of the bullfrogs were infected by Eustrongylides sp. with a mean intensity of infection of 13.4 specimens per host. Four host specimens (31%) presented pathological signs that seemed to be related to the Eustrongylides sp. infection, such as internal organs adhered to each other due to high intensity and large size of the nematodes, ulcers, and raw flesh wounds caused by the nematode. The pathogenic and concomitant infections have potential zoonotic implications and raise concerns about human infection risks for Eustrongylides infection. Moreover, such infections may represent an additional level of threat to native communities through the potential shifts in patterns of parasite and pathogen transmission. Future research involving the native anuran community is essential to ascertain whether invasive bullfrogs are attenuating or exacerbating diseases such as ranavirosis and eustrongylidiosis.

Nephrectomy in 52 dogs for the treatment of Dioctophyme renale infection - Knowing the enemy to win.

Dioctophyme renale (D. renale) is a nematode that parasitizes the kidney of mammals. Treatment is often surgical, with removal of the affected organ. This retrospective study aims to evaluate the epidemiological, clinical, and surgical aspects, the interval between diagnosis and treatment, the occurrence of pre- and intraoperative complications, and the postoperative survival time of dogs parasitized by D. renale undergoing therapeutic nephrectomy. Records of fifty-two dogs treated in a single hospital service were analyzed. We collected epidemiological data, laboratory results, diagnostic method, anesthetic protocol, surgical technique and time, type of antimicrobial prophylaxis, pre- and intraoperative complications, location and number of parasites, and postoperative survival time. Of the 52 dogs undergoing right nephrectomy by laparotomy, 61.5 % were female and 63.4 % were adults. Although the most common clinical sign was hematuria (25 %), 61.5 % of the patients were asymptomatic. Eosinophilia and increased serum urea were the only laboratory changes found. The interval between diagnosis and surgery was 27.4 ± 23 days and no patient showed changes suggestive of surgical emergency. The most common surgical approach was the right paracostal (61.5 %), and a continuous suture pattern was predominant. Intraoperative complications occurred in 9.6 % of the procedures, varying from mild to severe hemorrhage. Mean postoperative survival was 835.5 ± 428 days. Dioctophymosis was effectively controlled by nephrectomy of the affected kidney, allowing a mean survival of more than 830 days. No serious complications caused by intervals between diagnosis and treatment have been reported. This is the largest retrospective study evaluating dogs infected with D. renale that were surgically treated.

Tracking the existence of Dioctophyma renale in Parthian Empire of Iran (247 BC-224 AD).

Paleoparasitological investigation in the archeological site of the Caspian Sea littoral revealed the presence of human Dioctophyma renale infection, a ubiquitous kidney-residing helminth of mammals, during the Parthian Empire of Iran (247 BC-224 AD). This study reports the oldest human infection with giant kidney worm in the Middle East, mainly in an area where the infection is reported nowadays. The paucity of records throughout the literature suggests that the same species of parasite occurred with low frequency in the past, as well. The cemetery in the archeological site of Kiasar represented a unique opportunity of studying human and animal parasites of the Parthian Empire in Iran.

Identification of Larval Eustrongylides (Nematoda: Dioctophymatoidea) sp. from Channa punctata Bloch, 1793 by Morphological and Molecular Techniques

Objective: This study aimed to identify the larval form of Eustrongylides sp. isolated from the visceral organs of Channa punctata (Bloch, 1793) using morphological and molecular methods. Methods: Fishes were collected from fish farms in Nadia and North 24 Paraganas for the collection of nematodes. The visceral organs were dissected and kept in 0.67% normal saline. Nematodes collected from the abdominal regions and visceral organs for light microscopy study were fixed in 70% ethanol. Morphological features were studied by placing the nematodes in lactophenol. The specimens were later preserved in 70% ethanol containing 5% glycerine. Specimens processed for scanning electron microscopy were fixed in 2.5% glutaraldehyde and postfixed in 1% osmium tetraoxide. Proper identification was done by using standard methology. Molecular studies were performed for the 18S rRNA, ITS1, 5.8S rRNA, ITS2 and 28S rRNA gene fragments using polymerase chain reaction amplification, sequencing and phylogenetic analysis. Results: The morphological characteristics of nematodes were described with the help of light and scanning electron microscopy. Additional features not described earlier like dimensions and shape of the cephalic papillae, absence of somatic papillae, presence of caudal papillae, were identified for the first time. Moreover, molecular studies with ITS regions further confirmed the identification of the nematode. Conclusion: Thus the use of morphotaxonomy along with molecular techniques would help in proper identification of Eustrongylides sp infecting edible fish. Studies on the nematode would help toexplore the intermediate as well as paratenic hosts of the parasite. Data in this regard would contribute significantly to the fish database in regard to parasites infesting edible fishes.

OCCURRENCE OF EUSTRONGYLIDES EXCISUS (NEMATODA:DIOCTOPHYMATIDAE) IN EUROPEAN PERCH (PERCA FLUVIATILIS) AND GREAT CORMORANT (PHALACROCORAX CARBO) IN LAKE ANNONE, NORTHERN ITALY.

The genus Eustrongylides includes zoonotic nematodes that infect fish species and fish-eating birds of freshwater ecosystems. This study aimed to evaluate the occurrence of Eustrongylides in the paratenic host Perca fluviatilis (European perch) and in the definitive host, Phalacrocorax carbo sinensis (great cormorant), in Lake Annone, a shallow eutrophic lake located in the pre-mountainous area of the Alps in northwest Italy where wintering cormorants coexist with new breeding colonies. A total of 114 European perch and 48 cormorants were examined for the occurrence of Eustrongylides. All parasites collected were identified with microscopic examination and molecular analysis. Overall, 11 specimens of European perch (9.6%) and 13 individuals of cormorants (27%) harbored nematodes identified as fourth-stage larvae and adults of Eustrongylides excisus. The observed prevalence of Eustrongylides spp. appears to be intermediate between the higher values in cormorant breeding areas in northern Europe and the lower prevalence observed in their wintering sites in southernmost Europe. Considering the eutrophication status of freshwater ecosystems and the increasing population of the cormorants, Eustrongylides has an increasing potential range of dispersion in Europe, including Italy; thus an extensive surveillance should be carried out, especially given the zoonotic potential of this nematode.

REVIEW OF DIOCTOPHYME RENALE: ETIOLOGY, MORPHOLOGY, BIOLOGY, ECOEPIDEMIOLOGY, PATHOGENESIS, SYMPTOMATOLOGY, DIAGNOSIS, TREATMENT, AND PREVENTION.

Dioctophyme renale has a worldwide distribution and has been diagnosed in several wild and domestic animals as well as in humans. As numerous reports in the literature exist concerning the presence of D. renale in different animal species, as well as its diagnosis, treatment, and confirmation in new geographic areas, we reviewed existing information to contribute to the knowledge of the etiology, biology, epidemiology, pathogenesis, symptoms, diagnosis, treatment, and prevention of D. renale. Results of dioctophymosis may range from asymptomatic infection to even death of the host. Diagnosis is based on data from morphology, imaging, and antibody testing, with surgical treatment being the most effective. A high potential for infection of pets exists when there is overlap with wild parasitized animals; given common risk factors for infections in humans, D. renale should be considered as having zoonotic potential.

GIANT RED KIDNEY WORM (DIOCTOPHYMA RENALE) INFECTION IN PUPPIES LESS THAN FOUR MONTHS OF AGE FROM NORTHERN CANADA.

In this article, we present 2 case reports of puppies less than 4 mo of age at the time of infection with the nematode known as giant red kidney worm (Diotophyma renale). The first puppy was prepared for an ovariohysterectomy that evolved into an exploratory laparotomy. An inflamed omentum was observed, and D. renale infection was suspected due to the puppies' place of origin. The algorithm developed by the Ontario Society for the Prevention of Cruelty to Animals for the diagnosis and treatment of D. renale infections in dogs was followed. Both puppies had a history of origin from northern Manitoba, Canada. Due to the puppies' ages, the authors suspect that the transmission of infection for both puppies could be transmammary, transplacental, or ingestion of paratenic hosts or that these cases challenge what is thought to be the current prepatent period. In addition, D. renale infections in dogs, especially from northern communities, could be common.

Dioctophyme renale: in vitro culture, production of excretory and secretory antigens, and their use in immunodiagnosis.

The nematode Dioctophyme renale affects mainly the right kidney of domestic and wild mammals, in addition to having zoonotic potential. Therefore, this study aimed to produce excretion and secretion antigens of adult D. renale (DES) to diagnose canine dioctophimosis. To obtain the excretion and secretion antigens (DES), five D. renale adults were maintained for three weeks in supplemented RPMI medium with weekly supernatant collections. After the DES concentration, 2200 mL of the collected supernatant was used, which was centrifuged, followed by two filtrations and dialyzed, resulting in 12.5 mL of DES with a protein concentration of 0.59 mg/mL. The polyacrylamide gel electrophoresis (SDS-PAGE) of the DES antigen showed fractions with molecular weights ranging from 30 to 250 Kda. In the indirect ELISA with the DES antigen, the mean absorbance of the positive sera (38) was 0.839 ± 0.267, and the mean of the negative control sera (7) was 0.208 ± 0.083; the specificity and sensitivity of the ELISA were 100 and 97.4 %, respectively, being as effective as the surgical method in the presence of viable parasites. Thus, for the first time, this study describes the production of excretion and secretion antigens of adult D. renale and standardizes an indirect ELISA to diagnose dioctophimosis.

Dioctophyme renale in wandering dogs in Pelotas, South of Brasil.

This study aimed to verify the occurrence of Dioctophyme renale in stray dogs in the city of Pelotas, in the state of Rio Grande do Sul, Brazil. The Laboratório Regional de Diagnóstico of the Universidade Federal de Pelotas received 146 wandering dogs for necropsy, sent by the City Hall of Pelotas from March 2012 to January 2020. Among the necropsied animals, seventeen dogs (11.64%) were diagnosed with dioctophymosis. Among these dogs, 11 were parasitized with one specimen in the right kidney, two dogs presented two specimens in the right kidney, and in other two dogs, the parasites were in the abdominal cavity. In one dog, two parasites were found in the left kidney; in another dog, both kidneys were parasitized, with two parasites in the right kidney and one in the left kidney. The data obtained in this study showed that the occurrence of D. renale in stray dogs in the city of Pelotas is high, and D. renale mainly parasitizes the right kidney.

A rare case of ectopic dioctophymosis in Leopardus pardalis (Carnivora, Felidae) in Paraguay.

An adult female Leopardus pardalis from Jardín Botánico y Zoológico de Asunción, Paraguay was brought in from the wild, and inspected for a parasitic structure attached to the vulva. The parasite was removed and was identified as a Dioctophyme renale female. Blood samples and ultrasound did not show abnormalities in any organ, confirming the ectopic presentation of the parasite. This is the first report of D. renale in the L. pardalis and a first time for ectopic location in vagina and spontaneous elimination in a felid.

Dioctophyme renale in wandering dogs in Pelotas, South of Brasil / Dioctophyme renale em cães errantes no município de Pelotas, Sul do Brasil

Abstract This study aimed to verify the occurrence of Dioctophyme renale in stray dogs in the city of Pelotas, in the state of Rio Grande do Sul, Brazil. The Laboratório Regional de Diagnóstico of the Universidade Federal de Pelotas received 146 wandering dogs for necropsy, sent by the City Hall of Pelotas from March 2012 to January 2020. Among the necropsied animals, seventeen dogs (11.64%) were diagnosed with dioctophymosis. Among these dogs, 11 were parasitized with one specimen in the right kidney, two dogs presented two specimens in the right kidney, and in other two dogs, the parasites were in the abdominal cavity. In one dog, two parasites were found in the left kidney; in another dog, both kidneys were parasitized, with two parasites in the right kidney and one in the left kidney. The data obtained in this study showed that the occurrence of D. renale in stray dogs in the city of Pelotas is high, and D. renale mainly parasitizes the right kidney.

Resumo Este trabalho possui como objetivo verificar a ocorrência de D. renale em cães errantes do município de Pelotas, no Rio Grande do Sul, Brasil. O Laboratório Regional de Diagnóstico da Universidade Federal de Pelotas recebeu 146 cães errantes para necropsia, encaminhados pela Prefeitura Municipal de Pelotas, no período de março de 2012 a janeiro de 2020. Dentre os cães necropsiados, 17 animais foram diagnosticados com dioctofimose, determinando uma ocorrência de 11,64%. Desses animais, onze estavam parasitados com um exemplar no rim direito, dois animais possuíam dois exemplares no rim direito. Em dois animais, os parasitas estavam na cavidade abdominal; em um animal havia, dois parasitas no rim esquerdo e noutro animal os dois rins estavam parasitados, tendo dois exemplares no rim direito e um exemplar no rim esquerdo. Os dados obtidos neste trabalho permitem concluir que é elevada a ocorrência de D. renale nos cães errantes do município de Pelotas, parasitando principalmente o rim direito.

Occurrence of ectopic Dioctophyma renale in a Bolivian dog.

This is the first scientific report of the ectopic occurrence of Dioctophyma renale in subcutaneous tissue of a domestic dog's paw from Bolivia. An 18-month-old male dog (Pug) from Bolivia (San Matías) was examined on September 15, 2020 presenting limited mobility and apparent swelling in the left pelvic limb. After clinic evaluation, left foot edema was observed from tibio-tarsal to metatarsophalangeal joints which was more pronounced on the dorsolateral face. An incision at the site revealed serosanguinous exudation associated with a wide reddish nematode, which was identified as a female Dioctophyma renale. Epidemiological considerations were highlighted as this disease has zoonotic potential, and the infected dog was kept in a yard to which amphibians (Anura) and reptiles had access. Moreover, there is a lack of information on its occurrence in San Matías, a border city with Brazil.

Morphological and molecular characterization of Eustrongylides excisus larvae (Nematoda: Dioctophymatidae) in Sander lucioperca (L.) from Northern Turkey.

The genus Eustrongylides Jägerskiöld, 1909 includes parasitic nematodes (Dioctophymatidae) affecting various fish species and piscivorous birds of freshwater ecosystems. Currently, there is little information on the molecular characterization of E. excisus based on nuclear ribosomal internal transcribed spacer (ITS) rDNA regions. However, before the present study, there had been no reports of characterizing the E. excisus using nuclear small subunit ribosomal RNA (SSU rRNA) and mitochondrial cytochrome c oxidase subunit I (COI) genes sequences. In the present study, Eustrongylides spp. larvae were collected from pike-perch Sander lucioperca (L.) in Northern Turkey, and characterized by sequencing of ITS regions, SSU rRNA, and COI markers. Larvae herein morphologically identified as the fourth stage of Eustrongylides spp. were genetically identified as E. excisus based on the ITS sequence analysis. This study is the first record of SSU rRNA and COI sequences for E. excisus in GenBank. This is also a molecular characterization of E. excisus for the first time in Turkey. The ITS, SSU rRNA, and COI sequences of E. excisus can be used to establish the phylogenetic relationships of Eustrongylides species from Turkey and worldwide for further studies.

Dioctophimosis: A Parasitic Zoonosis of Public Health Importance.

Dioctophyme renale, the giant kidney worm, is a renal nematode from domestic and wild mammals that has zoonotic potential. In humans, dioctophimosis has been reported in several countries, mainly on the Asian continent, totaling more than 40 cases, which describe the parasite mainly infecting the kidneys, bladder, urethra and skin. Infection in animals and humans is related to the ingestion of the infective larva (L3) present in the aquatic oligochaete annelid (mandatory intermediate host) or fish and anurans (facultative paratenic hosts). Thus, the infection is related to the habit of drinking water contaminated with the mandatory intermediate host, as well as raw or undercooked meat from the facultative paratenic hosts. Dioctophimosis destroys the renal parenchyma and, in some cases, can cause the death of its hosts. In this chapter, we discuss the main topics regarding dioctophimosis in humans, domestic and wild animals, highlighting its importance in public health.

Occurrence of Dioctophyme renale (Goeze, 1782) in road-killed canids of Iran and its public health implication.

Dioctophyme renale, is the largest of parasitic nematodes, which infects different species of fish-eating carnivores worldwide. The northern provinces of Iran (Guilan and Mazandaran) located in south of the Caspian Sea are suitable for parasitic infections due to the mild and humid climatic conditions. From separate surveys of road-killed canids in various parts of the Caspian Sea littoral area in Iran, 70 carcasses were collected along the roads of Guilan and Mazandaran from 2015 to 2017. Dioctophyme renale detected by direct observation and molecular methods based on Cytochrome c oxidase subunit 1 (COX1 gene) sequencing analysis. Molecular investigation was also performed to validate prevalence and reduce false negative concerns. Dioctophyme renale was found in eight of 70 carnivores, mostly in the right kidneys, as well as two cases in the abdominal cavity of a dog and a golden jackal. More carcasses on the roads were seen with lacerated internal organs. Given the frequent number of giant kidney worms in canids in the region, the transmission of this zoonotic helminth to humans seems possible, since the area is a tourism hub in the country. The infection burden of this helminth should be investigated using DNA analysis of kidney tissue of road-killed carnivores in Iran.

Giant Red Kidney Worm (Dioctophyma renale) Screening and Treatment Protocol and Aberrant Worm Migration In Dogs From Ontario and Manitoba, Canada.

The life cycle of Dioctophyma renale involves an intermediate host (oligochaete), a paratenic hosts (fish and frogs), and a definitive host (mustelids and canids). Dogs are at risk of infection with D. renale when they consume paratenic hosts infected with the larval form of D. renale. Water containing the oligochaete intermediate host cannot be disregarded as another source of infection. Infections occur mainly in the right kidney, but worms have also been found in the abdominal cavity as well as other organs. Most dogs appear asymptomatic and infections are usually noted as incidental findings on necropsy. Recently, the Ontario Society for the Prevention of Cruelty to Animals (SPCA) and Humane Society conducted transports of dogs located in northern remote communities. In 2016, some female dogs were found to be infected with D. renale upon ovariohysterectomy. In response to this discovery, we developed a screening protocol to screen for D. renale infections. In 2018, a total of 130 intact dogs were transferred from 2 northern communities in the provinces of Ontario and Manitoba. A prevalence of 7.94% (95% confidence interval 3.87-14.11%) was found from dogs from the northern communities. The screening protocol we developed provides a method of screening for dogs that are transported from communities that could be at risk of infection with D. renale.