The impact of Anguillicoloides crassus (Nematoda) on European eel swimbladder: histopathology and relationship between neuroendocrine and immune cells.

The swimbladder functions as a hydrostatic organ in most bony fishes, including the European eel, Anguilla anguilla. Infection by the nematode Anguillicoloides crassus impairs swimbladder function, significantly compromising the success of the eel spawning migration. Swimbladders from 32 yellow eels taken from Lake Trasimeno (Central Italy) were analysed by histopathology- and electron microscopy-based techniques. Sixteen eels (50%) harboured A. crassus in their swimbladders and intensity of infection ranged from 2 to 17 adult nematodes per organ (6.9 ± 1.6, mean ± s.e.). Gross observations of heavily infected swimbladders showed opacity and histological analysis found a papillose aspect to the mucosa and hyperplasia of the lamina propria, muscularis mucosae and submucosa. Inflammation, haemorrhages, dilation of blood vessels and epithelial erosion were common in infected swimbladders. In the epithelium of parasitized swimbladders, many empty spaces and lack of apical junctional complexes were frequent among the gas gland cells. In heavily infected swimbladders, we observed hyperplasia, cellular swelling and abundant vacuolization in the apical portion of the gas gland cells. Numerous mast cells and several macrophage aggregates were noticed in the mucosal layer of infected swimbladders. We found more nervous and endocrine elements immunoreactive to a panel of six rabbit polyclonal antibodies in infected swimbladders compared to uninfected.

Gastric Physaloptera Infection in 27 Dogs (1997-2019).

Infection with the stomach worm Physaloptera spp. in dogs can cause chronic vomiting, although the diagnosis is often difficult owing to a low worm burden, single-sex infections, a failure to produce ova, or ova that are of greater density than solutions routinely used for qualitative fecal flotation. A retrospective evaluation was performed of 27 dogs that had gastric Physaloptera spp. infection confirmed endoscopically. In 23 of 27 dogs (85.2%), chronic vomiting was the chief complaint, and Physaloptera was an incidental finding in 3 dogs with esophageal or gastric foreign bodies. The worm burden was low (1-3 worms) in 21 dogs (77.8%), but 2 dogs were infected with large numbers (>50 worms). Prior therapy with routine doses of anthelmintics was ineffective in eight dogs prior to endoscopy. A higher dose and longer duration of fenbendazole in combination with pyrantel pamoate is recommended for treatment of suspected or confirmed infections. Reinfection is common in some dogs and should not be viewed as treatment failure.

PATHOLOGY OF SPIRURID INFECTION IN VERVET MONKEYS (CHLOROCEBUS PYGERYTHRUS) HOUSED IN A PRIMATE REHABILITATION CENTER.

Spirurids, specifically the Rictularia, Chitwoodspirura, Streptopharagus, and Protospirura genera, have been reported to parasitize all nonhuman primate taxa. Spirurid pathogenesis in nonhuman primates has not been reported frequently; however, Protospirura muricola has been associated with serious gastric pathologies, including gastric perforation. This study was a retrospective study of 38 vervet monkey (Chlorocebus pygerythrus) necropsies performed in a primate sanctuary that houses captive orphaned or injured wild-born vervet monkeys. Individuals were categorized according to their age, sex, and body condition score to investigate the relationships between these factors and parasite presence. This study identified P. muricola in 47.37% of the necropsied carcasses. Regarding individual factors associated with P. muricola infection, no significant differences between males and females were observed; however, relationships between parasite presence and poor body condition and advanced host age were observed. Furthermore, one monkey death was potentially directly related to spirurid pathogenic action, because the individual showed gastric perforation.

Aberrant Mesenteric Migration of Spirocerca lupi Larvae Causing Necrotizing Eosinophilic Arteritis, Thrombosis, and Intestinal Infarction in Dogs.

This report presents a novel canine condition in 32 dogs in which aberrant migration of Spirocerca lupi larvae through mesenteric arteries, instead of gastric arteries, led to small or large intestinal infarction. This form of spirocercosis was first recognized in Israel in 2013 and is currently ongoing. Typical clinical signs were anorexia and weakness of 3 to 4 days and, less frequently, vomiting and diarrhea, followed by collapse, bloody diarrhea, and severe vomiting. Exploratory laparotomy showed 1 or more infarcted and often perforated intestinal segments in all cases. Microscopically, there was intestinal mucosal to transmural coagulative necrosis and mesenteric multifocal necrotizing eosinophilic arteritis, thrombosis, hemorrhage, and early fibroplasia. Third-stage S. lupi larvae were identified by morphologic features in 9 of 32 (28%) cases, and the species was confirmed by polymerase chain reaction in 4 cases. Nearly 50% of the dogs had been receiving prophylactic therapy, which did not prevent this form of spirocercosis.

First report of Spirocerca vulpis in red foxes (Vulpes vulpes) in Portugal.

Recent studies have described Spirocerca lupi-like nematodes in the stomach of red foxes (Vulpes vulpes) in Europe. A phylogenetic analysis of those specimens using mitochondrial DNA and their morphological reexamination allowed their characterization as a different species, Spirocerca vulpis. Between the years of 2010 and 2017, roundworms were collected from seven red foxes of northeastern Portugal found at necropsy with nodular lesions on their stomach wall. Histopathological analysis of four foxes revealed granulomatous lesions of the gastric nodules. On morphological assessment, by light microscopy, nematodes revealed the presence of six triangular teeth-like buccal capsule structures, which are absent in S. lupi. Polymerase chain reaction was run to amplify a 551 bp partial fragment of the cytochrome c oxidase subunit 1 gene. Sequences were 99% similar to S. vulpis (85% coverage) of red foxes from Spain and Bosnia and Herzegovina, 99% similar (99% coverage) to sequences of Spirocerca sp. of red foxes from Denmark and 93% similar (99% coverage) to S. lupi from South Africa. This is the first report of S. vulpis in foxes or any other host from Portugal.

First record of the nematode Echinocephalus pseudouncinatus (Gnathostomatidae, Spirurida) in an edible, commercial host, the pen shell Atrina maura (Bivalvia: Pinnidae).

We describe a nematode infestation of the Atrina maura, the most valuable pen shell fished in Mexico, for the first time. Observations by Mexican authorities indicated that parasites were found encysted in pen shell gonads. The aims of this study included: (i) to identify the parasite morphologically; (ii) to quantify the infestation in different tissues of the pen shell; and (iii) to establish the seasonal variation in gonadal development. During 2015, 10 to 40 pen shells were randomly collected each month from the same commercial bed, and gonads were preserved and processed histologically to establish the seasonal and monthly frequency of infested gonads. A subsample of 35 pen shells was used to identify the parasite and to characterize the infestation of the muscle, mantle, gonad, and digestive gland tissue components. All tissues were dissected, and parasites were collected, photographed, quantified, and adequately preserved for species identification using scanning electron microscopy. Prevalence, intensity of infestation, abundance and crowding data were analyzed by Quantitative Parasitology software and compared monthly and seasonally using a chi test and analysis of means. Results indicated that all parasites were juvenile (second stage) nematodes, Echinocephalus pseudouncinatus. Parasites were found only in muscle and gonad tissues in both male and female pen shells. Hyperpigmentation was observed in the muscle and atretic oocytes with phagocytosis in the gonads. The frequency of infested gonads was significantly higher during winter but was not related to shell size or sex (p > 0.05). A greater number of pen shells were observed to be infested during December and August and during pen shell spawning and development stages. The highest frequency of infested gonads (40%) occurred in December when the lowest temperature and chlorophyll a concentration were recorded. The effects of the biotic and abiotic parameters on this host-pathogen relationship need further research. This nematode species is related to species that are causal agents of gnathostomiasis, which may constitute a health hazard for raw muscle consumers and could affect the reproductive activity of pen shell.

Habitat use pattern of the giant parasitic nematode Crassicauda magna within the pygmy sperm whale Kogia breviceps.

The giant (>3 m) parasitic nematode Crassicauda magna infects kogiid whales, although only 3 studies to date have provided detailed descriptions of these worms, all based upon fragmented specimens. These fragments were found within the neck region of kogiids, an unusual anatomic site for this genus of parasites. C. magna is a species-specific parasite among kogiids, infecting only pygmy sperm whales Kogia breviceps, and with a primarily cervico-thoracic distribution. To date, however, the pattern of habitat use within the host and transmission path of this parasite remain unknown. We used detailed dissections (n = 12), histological examination of host tissues (n = 2), and scanning electron microscopy of excised nematodes (n = 7) to enhance our understanding of this host-parasite relationship. Results revealed that a critical habitat for the parasite is an exocrine gland in the whale's ventral cervical region. C. magna male and female tails were found intertwined within the glandular lumen, and eggs were observed within its presumed secretion, illuminating the transmission path out of the host. The cephalic ends of these worms were often meters away (curvilinearly), embedded deeply within epaxial muscle. A single worm's complete, tortuous 312 cm course, from the gland to its termination in the contralateral epaxial muscle, is described for the first time. This study also provides the first scanning electron micrographs of C. magna, which illustrate taxonomically important features of the heads and tails of both male and female worms.

A new species of philometrid parasite (Nematoda, Philometridae) and histopathological lesions in juvenile whitemouth croakers, Micropogonias furnieri (Desmarest).

A new species of nematode parasite, Philometroides tahieli sp. nov. is described. The juvenile whitemouth croaker, Micropogonias furnieri Desmarest, was collected during the spring and summer of the years 2008-2010 from estuarial environments of Argentina. During investigations of the parasite fauna a nematode encapsulated in the musculature of the operculum was found. The nematodes were removed from the host's muscle tissue and slide-mounted in lactophenol solution to clarify the specimens. Opercular muscle with the parasite was processed for histopathological examination. Philometroides tahieli sp. nov. can be identified by the location of gravid females in the host, presence of anterior oesophageal bulb, bosses on the surface of the body, presence of transversal mounds and 14 cephalic papillae in two circular rows. The pathogenicity of the parasite is low in the natural environment, but lesions are consistent with a chronic process. The appearance of caseous necrosis suggests the presence of a locally acting substance. Philometroides tahieli sp. nov. is the second species of this genera described from brackish waters, and since the discovery of Philometroides maplestoni in 1928, is the first species of this genus recorded for South America.

Comparative imaging of European eels (Anguilla anguilla) for the evaluation of swimbladder nematode (Anguillicoloides crassus) infestation.

This study compares diagnostic imaging tools in detecting the parasitic swimbladder nematode Anguillicoloides crassus in Anguilla anguilla (L.) and focuses on ultrasound in an attempt to develop a non-destructive, field diagnostic test. Ultrasound use could allow the parasite to be diagnosed without decreasing the number of critically endangered European eels through post-mortem. In the preliminary study, eels were examined with computed radiography, computed tomography, magnetic resonance imaging, 14 MHz high-end ultrasound and 5 MHz low-end portable ultrasound, and the results were compared with post-mortem findings. This ultrasound scanning technique did not produce any promising results. A second batch of eels was examined using the same high-end and low-end ultrasounds, but employing a different scanning technique and comparing the results with post-mortem. This second study, scanning along the midline from below, allowed for the detection of anomalies associated with moderately infected animals. None of the eels used in this study were severely infected; thus, no conclusions can be made regarding the use of ultrasound in those animals. Overall, it was found that none of the techniques were useful in diagnosing mildly infected individuals; therefore, no single diagnostic imaging tool is sensitive enough to replace post-mortem for definite diagnosis.

A Pediatric Case of Thelaziasis in Korea.

In the present study, we intended to report a clinical pediatric case of thelaziasis in Korea. In addition, we briefly reviewed the literature on pediatric cases of thelaziasis in Korea. In the present case, 3 whitish, thread-like eye-worms were detected in a 6-year-old-boy living in an urban area and contracted an ocular infection known as thelaziasis incidentally during ecological agritainment. This is the first report of pediatric thelaziasis in Seoul after 1995.

The occurrence and pathogenicity of Serratospiculum tendo (Nematoda: Diplotriaenoidea) in birds of prey from southern Italy.

The air sacs of free-ranging birds of prey (n= 652) from southern Italy, including 11 species of Accipitriformes and six of Falconiforms, were examined for infections with Serratospiculum tendo (Nematoda: Diplotriaenoidea). Of the 17 species of birds examined, 25 of 31 (80.6%) peregrine falcons (Falco peregrinus) from Calabria Region and a single northern goshawk (Accipiter gentilis) from Campania Region were infected with S. tendo, suggesting a strong host specificity for the peregrine falcon. The northern goshawk and 18 of 25 infected peregrine falcons showed cachexia and all infected birds had bone fractures. At gross examination, air sacculitis and pneumonia were the most common lesions in infected birds. Microscopically, the air-sac walls showed thickening of the smooth muscle cells, resulting in a papillary appearance, along with hyperplasia of the mesothelium and epithelium, and foci of plasma cell infiltration and macrophages associated with several embryonated eggs and adult parasites. Extensive areas of inflammation were found in the lungs, characterized by lymphocytes, macrophages and fibroblasts surrounding embryonated eggs. The northern goshawk also had detachment of the dextral lung with several necrotic foci. In this case, the death of the bird was directly attributed to S. tendo infection. Lesions and pathological changes observed here suggest that S. tendo can cause disease.

Comparison of infection success, development and swim bladder pathogenicity of two congeneric Anguillicola species in experimentally infected Anguilla anguilla and A. japonica.

Two closely related parasites, Anguillicola crassus and Anguillicola novaezelandiae, originally parasitizing swim bladders of the Japanese eel (Anguilla japonica and the Short-finned eel (Anguilla australis), respectively, were used for analyzing the infection success of each parasite species on either long-known, recently acquired or new definitive host species and the associated effects on the eels' swim bladders. On that account, European eels (Anguilla anguilla) and Japanese eels were experimentally infected with both Anguillicola species in the laboratory. Susceptibility of the two eel species to both parasite species was determined by analyses of infection data. Subsequently, histopathological effects of the nematodes on the hosts' swim bladders were characterized according to already established indices.The present study revealed significant differences between the four different host-parasite systems regarding recovery rates, infrapopulations, and damage levels. Both nematode species achieved significantly lower recovery rates in Japanese eels than in European eels, since the examined swim bladders of Japanese eels contained a high amount of dead encapsulated larvae, whereas those of European eels contained only living nematodes. Encapsulation of larvae in Japanese eels was associated with a distinct thickening of the swim bladder walls. The swim bladders of uninfected Japanese eels turned out to be generally thicker than those of European eels. Infection with both Anguillicola species resulted in a further thickening process of the swim bladder walls in Japanese eels, whereas those of European eels showed only minor changes. The two established classification systems turned out to be inapplicable, since the measurements and the macroscopic evaluations of the swim bladders of the two infected eel species did not entirely correspond to the underlying criteria.

Fatal verminous pharyngitis and esophagitis caused by Streptocara incognita in mute swans (Cygnus olor).

Streptocara spp. infections are reported to cause gastritis, proventriculitis, esophagitis, and pharyngitis in various waterfowls, especially diving ducks. In the present paper, we describe severe fatal diphtheritic pharyngitis and esophagitis caused by Streptocara incognita in three female mute swans (Cygnus olor) in Bosnia and Herzegovina. Prior to death, the swans were showing signs of lethargy, anorexia, and reluctance to move. At necropsy, in all swans severe diphtheritic pharyngitis and esophagitis with deep, dark red hemorrhagic ulcerations were observed. Numerous thin, white, up to 1-cm-long nematodes, identified as S. incognita, were observed embedded in the pharyngeal and esophageal mucosa under the diphtheritic membranes. Histopathology revealed severe fibrinonecrotic inflammation with numerous cross-sections of the parasites. To the authors' knowledge, this is the first report of severe, fatal streptocariasis in mute swans.

Canine ocular thelaziosis caused by Thelazia callipaeda in Portugal.

Ocular thelaziosis caused by Thelazia callipaeda is a vector-borne disease affecting dogs and humans. We report a case of thelaziosis in a 10-year-old German Shepherd dog from Vila Real city (Portugal). Ophthalmological examination revealed bulbar and nictitating membrane conjunctival hyperemia with serous discharge noted at the left medial canthus and blepharitis. Schirmer tear test value and intraocular pressure were within the reference ranges in both eyes, and biomicroscopy showed a transparent cornea without lesions or edema and no inflammatory reaction in the anterior chamber. No funduscopic alterations were detected by direct and indirect ophthalmoscopic examination. When testing the nasolacrimal patency, two white worms were observed on the caruncle conjunctival surface with undulating movements that increased with light intensity. In total, eight worms were collected and morphologically identified as T. callipaeda (seven mature females and one male). PCR amplification of a 689 sequence of partial cytochrome c oxidase subunit 1 target gene confirmed the nematodes were T. callipaeda, haplotype 1. The dog was treated with a single subcutaneous injection of ivermectin combined with additional topical application of ophthalmic fusidic acid drops and oral milbemycin oxime. One week after treatment, no worms were detected and the ocular clinical signs resolved. The most recent reports of canine thelaziosis in the Iberian Peninsula should alert local health authorities to the zoonotic potential of infestation with T. callipaeda, which should be included in the differential diagnosis of conjunctivitis in dogs and humans.

Prevalence of Anguillicoloides crassus and growth variation in migrant yellow-phase American eels of the upper Potomac River drainage.

Prevalence of the non-native swim bladder nematode Anguillicoloides crassus has recently increased in American eels from estuaries of the North American Atlantic coast, but little is known about parasite prevalence or conditions of previous infection in upstream migrant eels within upper watersheds. This study is the first to confirm presence of A. crassus in the upper Potomac River watershed. We estimated A. crassus prevalence during 3 time periods: September to October 2006 (5/143 eels, 3.5%), August to October 2007 (0/49 eels), and June 2008 (0/50 eels). All eels were sampled from the Millville Dam eel ladder on the lower Shenandoah River, a Potomac River tributary located approximately 285 km upstream of Chesapeake Bay, USA. Of the 5 infected eels, parasite intensity was 1 for each eel, and mean intensity was also 1.0. A swim bladder degenerative index (SDI) was calculated for the 50 eels from the final sampling period, and 38% of those eels (19 of 50) showed signs of previous infection by A. crassus. We also aged 42 of the 50 eels (mean ± SE = 6.7 ± 0.29 yr, range 4 to 11 yr) from the final sampling period. Based on the range of possible SDI scores (0 to 6), severity of previously infected swim bladders was moderate (SDI = 1 or 2). Previously infected eels, however, had a lower length-at-age than that of uninfected eels. Female yellow-phase eels in upper watersheds develop into large highly fecund silver-phase adults; hence, a parasite-induced effect on growth of yellow-phase eels could ultimately reduce reproductive potential.

Non-invasive diagnosis for Philometra ovata (Nematoda) infection in the common minnow Phoxinus phoxinus.

Non-invasive diagnoses for most of the extraintestinal helminth infections among vertebrates are rare. In the present study, we developed and tested a non-invasive visual diagnosis method to detect the infection of the nematode Philometra ovata, which is parasitising in the body cavity of the common minnow Phoxinus phoxinus. By observing the surface of minnow's abdomen, we diagnosed infected minnows with irregular or vermiform-shaped uplifts as a consequence of the presence of P. ovata in the body cavity. We conducted the diagnosis in minnows with or without anaesthetisation, and our results showed the non-invasive method is highly reliable (correct rate of diagnoses > 95 %) in both anaesthetised and non-anaesthetised groups. The correct rate of diagnoses in truly uninfected fish (i.e. specificity) was nearly 100 % in both groups, while the correct rate of diagnoses in truly infected fish (i.e. sensitivity) was 71 and 65 % in anaesthetised and non-anaesthetised fish, respectively. The correct rate in fish diagnosed as infected or uninfected (i.e. positive or negative predictive power) of non-invasive diagnosis was nearly 100 % among the anaesthetised fish, and over 95 % in non-anaesthetised group. The diagnosis also yielded prevalence of P. ovata infection similar to the real prevalence in anaesthetised fish. Diagnoses conducted by an inexperienced observer indicated that the method is repeatable. Taken together, the present non-invasive method seems to be a promising new tool for non-invasive detection of P. ovata infections in minnows and probably can be applied for the detection of other body cavity dwelling metazoan parasites in various host taxa.

The first human case of Thelazia callipaeda infection in Vietnam.

A 26-year-old man residing in a village of Thai Nguyen Province, North Vietnam, visited the Thai Nguyen Provincial Hospital in July 2008. He felt a bulge-sticking pain in his left eye and extracted 5 small nematode worms by himself half a day before visiting the hospital. Two more worms were extracted from his left eye by a medical doctor, and they were morphologically observed and genetically analyzed on the mitochondrial cytochrome c oxidase 1 gene. The worms were 1 male and 1 female, and genetically identical with those of Thelazia callipaeda. By the present study, the presence of human T. callipaeda infection is first reported in Vietnam.

Immunohistochemical characterization of lymphocyte and myeloid cell infiltrates in spirocercosis-induced oesophageal nodules.

Spirocerca lupi is a nematode that infects the dog's oesophagus and promotes the formation of an inflammatory fibroblastic nodule that progresses to sarcoma in approximately 25% of cases. Spirocercosis-associated oesophageal sarcoma is an excellent and under-utilized spontaneous model of parasite-associated malignancy. The inflammatory infiltrate of paraffin-embedded, non-neoplastic oesophageal nodules (n = 46), neoplastic nodules (n = 25) and normal oesophagus (n = 14) was examined by immunohistochemistry using MAC387 (myeloid cells), CD3 (T cells), Pax5 (B cells) and FoxP3 (T regulatory cells) antibodies. Myeloid cells predominated in 70% of nodules, in pockets around the worms' migratory tracts and in necro-ulcerative areas in neoplastic cases. T cells predominated in 23% of cases with a focal or diffuse distribution, in the nodule periphery. No significant differences were observed between neoplastic and non-neoplastic stages. FoxP3+ cells were observed in low numbers, not significantly different from the controls. The inflammation in spirocercosis is characterized by pockets of pus surrounded by organized lymphoid foci. There was no evidence of a local accumulation of FoxP3+ cells, unlike many previous studies that have reported an increase in FoxP3+ T cells in both malignancies and parasite infections. The triggering factor(s) driving the malignant transformation of the spirocercosis-associated chronic inflammatory nodule warrants further investigation.