Echinocephalus inserratus sp. n. (Nematoda: Gnathostomatidae) from the stingray Pastinachus ater (Dasyatidae) and new records of congeneric and some other nematode larvae from teleost fishes off New Caledonia.

Based on light and electron microscopical studies, a new nematode parasite, Echinocephalus inserratus sp. n. (Spirurida: Gnathostomatidae), is described from the spiral valve of the broad cowtail stingray Pastinachus ater (Macleay) (Dasyatidae, Myliobatiformes) from off New Caledonia. The new species is morphologically and biometrically most similar to Echinocephalus overstreeti Deardorff et Ko, 1983, differing from it mainly in the absence of serrations on the posterior parts of pseudolabia and on interlabia, and in having a longer gubernaculum (150-299 µm long). Morphologically unidentifiable, mostly encapsulated larvae of Echinocephalus spp. were recorded from the following six species of teleost fishes collected in New Caledonian waters, serving as paratenic hosts: Perciformes: Acanthopagrus berda (Forsskål) (Sparidae) and Nemipterus furcosus (Valenciennes) (Nemipteridae); Tetraodontiformes: Abalistes stellatus (Anonymous), Pseudobalistes fuscus (Bloch et Schneider) (both Balistidae), Lagocephalus sceleratus (Gmelin) (Tetraodontidae) and Aluterus monoceros (Linnaeus) (Monacanthidae). Co-parasitising larvae of Ascarophis sp. and Hysterothylacium sp. were also collected from P. fuscus. All these findings represent new host and geographical records. A key to valid species of Echinocephalus Molin, 1858 is provided.

The occurrence and clinical importance of infectious stage of Echinocephalus (Nematoda: Gnathostomidae) larvae in selected Australian edible fish.

Cases of gnathostomiasis, an infection caused by consuming infected seafood, have been reported in Australia. However, doubt exists over the validity of these diagnoses as there are no reports of Gnathostoma spp. in Australian teleost fish. Also, the diagnoses in human cases were based on a serological test developed in Thailand. The specificity and sensitivity of this test in non-endemic areas are uncertain. Interestingly, parasites belonging to the genus Echinocephalus, which morphologically are very similar to Gnathostomum, are commonly found in Australian fish and shellfish and can potentially infect humans. The aim of this study was to determine the occurrence of these zoonotic nematodes within commercial fish and to characterise nematode larvae in order to provide insights into the specific identity of the potential causative agents of gnathostomiasis in Australia. Six edible fish species (n = 163) were examined. Gnathostomid-type larvae were found only in Acanthopagrus australis and Rhabdosargus sarba. Detailed examination and sequence data suggested parasite larvae belonged to the genus Echinocephalus. Further investigation of the occurrence of zoonotic nematodes within marine environments and observation of their spatial and temporal patterns will help raise awareness of the significance of this food safety issue within global fishing industries and health sectors. The accurate identification of zoonotic nematodes is a key component of disease surveillance and control. This information can also be used to develop specific and sensitive diagnostic test.

Secretome of the carcinogenic helminth Spirocerca lupi reveals specific parasite proteins associated with its different life stages.

Spirocerca lupi is a parasitic and carcinogenic nematode of canids distributed in tropical and subtropical regions around the world. The excretion-secretion proteins (PES) of S. lupi have been suggested to play a role in the pathogenesis of its infection. We aimed to identify the PES of different stages of S. lupi and search for proteins that would be useful for diagnostic, therapeutic and vaccination purposes as well as understand their functions. A nano-UPLC mass spectrometry de novo analysis was performed on proteins collected from cultures of S. lupi L3 larvae, L4 females, adult females and adult males from naturally infected hosts. A total of 211 proteins were identified in all cultures. Accordingly, 117, 130, 99 and 116 proteins were detected in L3 larva, L4 females, adult females and adult males, respectively, with a strong correlation in the biological replicates (Pearson coefficients > 0.73). Fourty-four proteins were detected in all developmental stages, 64 were stage-specific and 49 were exclusively identified in L4 females. Cell compartment enrichment analysis revealed that proteins common to all stages were cytoplasmatic (p < 9.x10-6), whereas L4 unique proteins were in collagen trimers, and macromolecular complexes (p < 0.00001). Functional enrichment analysis of proteins showed significant enrichment in lipid metabolism in L3-unique proteins (p<0.00005), in mannose metabolism and protein de-glycosylation for L4-unique proteins (p < 0.00004), and in phosphorus metabolism in proteins shared by all stages (p <  2.1 x10-9). Interestingly, annexin 6, associated with cancer in humans, was detected in all life stages, but in a larger abundance in L4 females and adults. These findings indicate that S. lupi establishes complex interactions with its hosts by an arsenal of proteins expressed in different patterns in each life stage which influence the pathogenesis and oncogenesis of S. lupi and may be used as potential targets for diagnostic assays, drug targets or vaccine candidates.

Life-cycle of Oxyspirura petrowi (Spirurida: Thelaziidae), an eyeworm of the northern bobwhite quail (Colinus virginianus).

BACKGROUND: Oxyspirura petrowi (Spirurida: Thelaziidae), a heteroxenous nematode of birds across the USA, may play a role in the decline of the northern bobwhite (Colinus virginianus) in the Rolling Plains Ecoregion of West Texas. Previous molecular studies suggest that crickets, grasshoppers and cockroaches serve as potential intermediate hosts of O. petrowi, although a complete study on the life-cycle of this nematode has not been conducted thus far. Consequently, this study aims to improve our understanding of the O. petrowi life-cycle by experimentally infecting house crickets (Acheta domesticus) with O. petrowi eggs, feeding infected crickets to bobwhite and assessing the life-cycle of this nematode in both the definitive and intermediate hosts. METHODS: Oxyspirura petrowi eggs were collected from gravid worms recovered from wild bobwhite and fed to house crickets. The development of O. petrowi within crickets was monitored by dissection of crickets at specified intervals. When infective larvae were found inside crickets, parasite-free pen-raised bobwhite were fed four infected crickets each. The maturation of O. petrowi in bobwhite was monitored through fecal floats and bobwhite necropsies at specified intervals. RESULTS: In this study, we were able to infect both crickets (n = 45) and bobwhite (n = 25) with O. petrowi at a rate of 96%. We successfully replicated and monitored the complete O. petrowi life-cycle in vivo, recovering embryonated O. petrowi eggs from the feces of bobwhite 51 days after consumption of infected crickets. All life-cycle stages of O. petrowi were confirmed in both the house cricket and the bobwhite using morphological and molecular techniques. CONCLUSIONS: This study provides a better understanding of the infection mechanism and life-cycle of O. petrowi by tracking the developmental progress within both the intermediate and definitive host. To our knowledge, this study is the first to fully monitor the complete life-cycle of O. petrowi and may allow for better estimates into the potential for future epizootics of O. petrowi in bobwhite. Finally, this study provides a model for experimental infection that may be used in research examining the effects of O. petrowi infection in bobwhite.

Thelaziosis due to Thelazia callipaeda in Europe in the 21st century-A review.

Thelazia callipaeda was first described at the beginning of the 20th century in Asia, but this eyeworm is now frequently reported in Europe in the 21st century. To date, thelaziosis has been described in the following European countries (in order of appearance): Italy, France, Germany, Switzerland, Spain, Portugal, Belgium, Bosnia and Herzegovina, Croatia, Serbia, Romania, Greece, Bulgaria, Hungary, Slovakia, the United Kingdom, Turkey and Austria. The infected vertebrate host species include domestic carnivores (dogs and cats), wild carnivores (red foxes, wolves, beech martens, wildcats and golden jackals), lagomorphs (brown hares and wild European rabbits) and humans. In Europe, 11 cases of human thelaziosis have been reported, the majority of which are autochthonous. However, some of them have been imported, a fact which highlights the importance of surveillance policies to restrict cross-border spread of the parasite. The objectives of this article are to review key aspects of the epidemiology of T. callipaeda, summarise animal and human cases in Europe and emphasise the importance of education and awareness among veterinarians, physicians (particularly ophthalmologists) and animal, in order to owners to tackle this zoonosis.

The first feline and new canine cases of Thelazia callipaeda (Spirurida: Thelaziidae) infection in Hungary.

BACKGROUND: In Europe, the first Thelazia callipaeda infections were found in the eyes of some dogs in Italy three decades ago. Since that time, this vector-borne nematode species has been diagnosed in domestic and wild carnivores and humans in some western European countries. During the last few years, autochthonous thelaziosis of dogs, red foxes, cats and humans has also been reported from eastern Europe. The first cases of ocular infections caused by T. callipaeda have been described in dogs living in the eastern and southern part of Slovakia and Hungary. METHODS: Whitish parasites found in the conjuctival sac and/or under the third eyelid of one or both eyes of animals were removed and morphologically identified according to species and sex. To confirm the morphological identification with molecular analysis a single step conventional PCR was carried out. RESULTS: A total of 116 adult worms (1-37 per dog, median: 7, IQR: 14.5 and 7 from a cat) were collected from the eyes of 11 animals. Nematodes were identified as T. callipaeda according to the morphological keys and molecular analysis. The sequences of a portion of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene were identical to those representing T. callipaeda haplotype 1, previously reported in neighbouring and other European countries. Since the infected cat and dogs had never travelled abroad, all of the cases were autochthonous thelaziosis. CONCLUSIONS: The present study reports the first case of thelaziosis in a cat and new cases in 10 dogs found in the southern and northern region of Hungary, respectively. Further studies are needed to clarify whether wild carnivores (e.g. red foxes, golden jackals) may act as reservoirs of this eyeworm species in the country.

The use of primary murine fibroblasts to ascertain if Spirocerca lupi secretory/excretory protein products are mitogenic ex vivo.

BACKGROUND: Spirocerca lupi is a nematode that parasitizes vertebrates in particular canids, by forming nodules in the thoracic cavity specifically in the oesophagus. In 25% of Spirocerca infections of the domestic dog, nodules progress from inflammatory to pre-neoplastic to sarcomatous neoplasia. With the mechanism of neoplastic transformation being incompletely understood, this study investigates if S. lupi parasite proteinaceous secretory/excretory products (ESPs) play a role in the neoplastic transformation. METHODS: To facilitate collection of ESPs, we maintained naturally harvested adult parasites in the laboratory under artificial conditions. Media in which the parasites were grown was subsequently evaluated for the presence of proteinaceous compounds using a mass spectroscopy library as well as for their ability to be mitogenic in primary murine fibroblastic cells. RESULTS: Chromatrography of the ethyl acetate extracted incubation media showed the presence of 9 protein compounds, of which three were identified as non-specific proteins isolated from Nematostella vectensis, Caenorhabditis brenneri and Sus scrofa, with the rest being unknown. Acetone, methanol, hexane and ethylacetate extracted culture media were unable to induce a mitogenic change in primary murine fibroblasts in comparison to the controls. CONCLUSION: While no mitogenic effect was evident, further studies are required to understand the role of worm excretory/secretory products on clastogenesis under chronic exposure. In addition, while not of primary importance for this study, the observed duration of parasite survival indicates that ex vivo studies on S. lupi are possible. For the latter we believe that the worm culture method can be further optimized if longer survival times are required.

In vitro lectin binding to the outer surface of Spirocerca lupi at different life-stages.

Spirocerca lupi is the esophageal nematode of dogs. Early, transient eosinophilia occurs in experimentally infected dogs, but is absent in advanced cases, suggesting that the nematode evades the dog's immune system. Lectins are proteins or glycoproteins of plant or animal origin, binding different saccharides, with varying specificities and avidities, used to characterize surface haptens in plant and animal parasitic helminths. This study investigated the in vitro binding of six lectins (Concanavalin A [ConA], wheat germ agglutinin [WGA], peanut agglutinin [PNA], soybean agglutinin [SBA], Dolichus biflorus agglutinin [DBA] and Ulex earopaeus agglutinin I [UEA]) to the surface of S. lupi nematodes at different life stages, the L2 and L3 larvae (dead and alive) and to dead adult worms, with negative controls, with and without addition of the six respective inhibitory sugar haptens. Con A moderately bound to surfaces of both live and frozen L3, to the stoma and excretory pores of adult worms, and to the outer surface nematode's eggs, within a female worm, but not to L2. PNA bound only to stoma and excretory pores surfaces in both frozen and live L3. WGA bound strongly to the outer surfaces of live and dead L2 and L3, which resulted in molting of live larvae. These results suggest that the nematode's surface content change during its development. Such changes may play roles in the nematode's interactions with the intermediate and definitive hosts' tissues, and in its ability to evade the immune response, its long survival within the host, and even induce neoplastic transformation.

Live eyeworm (Oxyspirura petrowi) extraction, in vitro culture, and transfer for experimental studies.

Northern bobwhite ( Colinus virginianus ) have experienced a dramatic decline in West Texas over the last 3 yr, and investigations are underway to evaluate the role of parasites in this decline. One of the key parasites being investigated is the eyeworm (Oxyspirura petrowi). Live eyeworms were extracted from both live and dead northern bobwhites, and in vitro survival was tested using 10 liquid media. Eyeworms placed in an egg white and physiological saline solution lived for at least 36 days. Live O. petrowi placed into the eyes of uninfected pen-raised bobwhites were monitored for 21 days to demonstrate successful transfer. Eyeworm behavior during feeding, mating, and development were monitored. This study is important to research that requires "banking" of live O. petrowi from wild-captured definitive hosts for life history studies and assessing the impact of O. petrowi on host individuals.

Serum C-reactive protein concentration in benign and malignant canine spirocercosis.

BACKGROUND: Spirocerca lupi is a nematode of canids that forms a nodule in the esophagus that can undergo neoplastic transformation. C-reactive protein (CRP) is a major acute phase protein in the dog that has been used for treatment, monitoring, and prognostication in inflammatory and neoplastic disease. HYPOTHESIS/OBJECTIVES: The objectives of this study were to determine if serum CRP concentration (1) is increased in canine spirocercosis, (2) can be used to determine neoplastic transformation, and (3) can be used to monitor response to treatment in benign spirocercosis. ANIMALS: Forty-two dogs naturally infected with S. lupi and 21 control dogs. METHODS: A retrospective study was performed. The infected cases were divided into benign (n = 28) or malignant (n = 14) spirocercosis. CRP was performed on all of the spirocercosis and control cases at presentation. Statistical analysis was done by the one-way analysis of variance and Student's t-test. RESULTS: The mean CRP concentration in the benign cases was 60.4 ± 48.0 mg/L and that of the malignant cases was 76.5 ± 44.8 mg/L; both values were significantly higher (P < .001) than those of the control group where the mean was 13.4 ± 17.9 mg/L. The mean CRP concentration for the convalescent sera in the benign group was lower than the pretreatment concentrations (P= .01). CONCLUSION AND CLINICAL IMPORTANCE: CRP cannot be used to differentiate between benign and malignant spirocercosis. There is a decrease in CRP concentration in dogs with benign spirocercosis once treatment has commenced. Serial CRP measurement can be used to monitor response to treatment in benign spirocercosis.

Experimental infection of selected arthropods with spirurid nematodes Spirocerca lupi Railliet & Henry, 1911 and Gongylonema ingluvicola Molin, 1857.

Gongylonema ingluvicola and Spirocerca lupi are spirurid nematodes that require arthropod intermediate hosts in order to complete their life cycle. Beetles of the family Scarabaeidae are reported to serve as intermediate hosts for both these parasites. In this study selected species of beetles of the family Scarabaeidae as well as other groups of arthropods were screened for susceptibility to infection with S. lupi and G. ingluvicola. Arthropods were exposed to infective eggs of both parasites for a determined period of time and dissected/digested to determine the presence or absence of pre-infective and infective larvae. All the five species of dung beetles exposed to infection with S. lupi, namely, Pachylomerus femoralis, Scarabaeus rugosus, Gymnopleurus humanus, Kheper nigroaeneus and Anachalcos convexus were susceptible and, of the two species exposed to G. ingluvicola, only Gy. humanus was susceptible. Spirocerca lupi eggs developed in millipede species, Daratoagonus cristulatus, and remained as encysted larvae, while in Orthoporoides kyrhocephalus no development was observed. Spirocerca lupi larvae were not detected in the cricket species Gryllus assimilis, or the cockroach species Periplaneta americana, and, similarly, G. ingluvicola larvae were not detected in the millipede species O. kyrhocephalus. The difference in the susceptibility of the arthropods to the two parasite species may depend on their feeding biology.

Prevalence of the dog nematode Spirocerca lupi in populations of its intermediate dung beetle host in the Tshwane (Pretoria) Metropole, South Africa.

Spirocerca lupi (Spirurida: Spirocercidae) is a cosmopolitan parasite, principally of domestic dogs and dung beetles are its main intermediate hosts. In South Africa there has recently been growing concern over the upsurge of reported cases of clinical spirocercosis in dogs, while little is known or understood about the dynamics of the host-parasite associations between dung beetles and this nematode. We determined and compared the prevalence of infection in dung beetles between rural, urban and peri-urban areas of Tshwane (Pretoria) Metropole. Dung beetles were sampled during April and October 2006, at various localities in each of these areas. Localities were selected on the basis of being focal areas of high infection with S. lupi in dogs. Pig, dog and cow dung-baited pitfall traps were used for sampling the beetles. Trap contents were collected 48 h after the traps had been set and only dung beetles were collected from the traps. In total, 453 specimens belonging to 18 species were collected from 63 pitfall traps in all three areas. The numbers of species that were collected varied among the three areas. Dung beetles, irrespective of species (18) and numbers (447), predominantly preferred pig dung. The prevalence of dung beetles infected with the larvae of S. lupivaried considerably in the three areas. In the urban area 13.5% of the dung beetles dissected were infected, while the prevalence of S. lupi in dung beetles in the rural area was 2.3%. All the dung beetles that were infected with this nematode showed a preference for omnivore (pig and dog) dung.

Human thelaziosis--a neglected parasitic disease of the eye.

The oriental eyeworm, Thelazia callipaeda (Spirurida, Thelaziidae), infects a range of definitive hosts, such as dogs, cats, foxes, rabbits, and humans. This parasite usually lives under the nictitating membrane of the eye, where the adult females release first-stage larvae into the lachrymal secretions; these larvae are subsequently ingested by the intermediate arthropod host within which they develop to the infective, third-stage larvae. The latter larvae are then deposited into the eyes of the definitive host. Recently, T. callipaeda has been reported to infect dogs, foxes, and/or cats in Europe (Italy, France, and Germany). Human thelaziosis (HT) is considered to be an underestimated parasitic disease, whose prevalence appears to have increased in poor socioeconomic settings in many Asian countries, including China. In humans, the disease can be subclinical or symptomatic, exhibiting epiphora, conjunctivitis, keratitis, excessive lachrymation, corneal opacity, and/or ulcers. Knowledge about HT is presently fragmentary and mainly limited to clinical case reports. This article provides a background on the parasite and its life cycle, reviews cases of human thelaziosis, summarizes key aspects regarding the diagnosis of thelaziosis, and proposes future research and methods of control of the disease in humans, particularly in Asia.

Nematode biology and larval development of Thelazia callipaeda (Spirurida, Thelaziidae) in the drosophilid intermediate host in Europe and China.

Thelazia callipaeda, commonly known as the 'oriental eyeworm', has been recently reported in Italy and other European countries. The insect/s that act as intermediate hosts and details of larval development inside the vector remain unclear. In order to (1) demonstrate the species of fly that may act as vector/s for T. callipaeda in southern Italy (Site A) and China (Site B) and (2) describe the larval development of the nematode in the body of flies, 847 Phortica (Drosophilidae) flies were collected from the above two sites, each with a history of human and/or canine thelaziosis. Flies were identified as Phortica variegata (245 - site A) and Phortica okadai (602 - site B), experimentally infected by 1st-stage larvae (L1), kept at different temperatures and dissected daily until day 180 post-infection (p.i.). Dead flies from site A were subjected to specific polymerase chain reaction (PCR) assay to detect T. callipaeda. To demonstrate the role of Phortica as vectors of T. callipaeda, 3rd-stage larvae (L3) recovered from the proboscis of flies were deposited onto the cornea of the eyes of dogs and rabbits. Following dissection, 3 (2.9%) of P. variegata in site A were found to be infected by L3 in the proboscis on days +14, +21 and +53 p.i., compared with 26 (18.4%) of Phortica flies recorded as being positive by PCR. Sequences from positive PCR products were 99% identical to sequences of the corresponding species available in GenBank (AY207464). At site B, 106 (17.6%) of 602 dissected P. okadai were found to be infected by T. callipaeda larvae (different stages) and in total 62 L3 were recovered from the proboscis of 34 (5.6%) flies. The shortest time in which L3 were found was at day +14, +17, +19, and +50 p.i. respectively, depending on the environmental temperatures. Of 30 flies overwintered for 6 months, 6 L3 were detected at day +180 p.i. in 3 flies (10%). The biology of larval development was reconstructed on the basis of the dissection of 602 P. okadai-infected flies and the morphology of larval stages in the insect body described. The present work provides evidence that P. variegata and P. okadai act as vectors for T. callipaeda in southern Europe and in China, respectively. The phenomenon of overwintering is described here for the first time for T. callipaeda and discussed. Finally, the relationship between T. callipaeda and its fly vector is considered in light of disease prophylaxis and to model its dissemination into habitats and environments favourable to Phortica flies.

Treatment of dog thelaziosis caused by Thelazia callipaeda (Spirurida, Thelaziidae) using a topical formulation of imidacloprid 10% and moxidectin 2.5%.

Canine thelaziosis caused by Thelazia callipaeda infects dogs, cats, foxes, rabbits, and humans resulting in conjunctivitis, pain and excessive lacrimation. T. callipaeda live in the eyes under the nictitating membrane and females release first stage larvae which are ingested by flies which act as intermediate hosts. Control of canine thelaziosis is currently based on the removal of nematodes directly from the eyes of affected dogs or on the local instillation of antiparasitic drugs. With the aim of evaluating the efficacy against T. callipaeda of an association of imidacloprid 10% and moxidectin 2.5% by spot-on formulation administered via dermal application, three groups of naturally infected animals were selected: i.e. group A (21 dogs) received a single dose of imidacloprid 10% and moxidectin 2.5% by spot-on; group B (21 dogs) received a single dose of imidacloprid 10% by spot-on and group C (20 dogs) were left untreated. The efficacy of treatments was established by eye inspection and parasite viability and vitality after 1, 5 and 9 days after animal treatments (groups A and B) and after 9 days only in untreated animals (group C). Imidacloprid 10% and moxidectin 2.5% in spot-on formulation showed to be effective with regards the control of dog thelaziosis within 5 (90.47%) to 9 (95.23%) days after treatment. Only one dog from group A presented nematodes after treatment. The presence of parasites in the eyes of dogs from groups B (imidacloprid 10%) and C confirm that the anthelmintic efficacy against T. callipaeda in animal from group A (imidacloprid 10% and moxidectin 2.5%) was most likely attributable to moxidectin 2.5%. The spot-on formulation containing imidacloprid 10% and moxidectin 2.5% is very easy to apply and helps overcome problems linked to the mechanical removal of parasites or to the restraining of the animals for the local instillation of drugs in the eyes.

A comparison between ethanol-induced chemical ablation and ivermectin plus prednizolone in the treatment of symptomatic esophageal spirocercosis in the dog: a prospective study on 14 natural cases.

This study included a total of 14 dogs with spontaneous esophageal spirocercosis. Historical and clinical evidence of esophageal dysphagia, detection of parasitic ova in fecal samples and endoscopic documentation of esophageal nodules were the inclusion criteria. The animals were randomly assigned into two groups: group A (n = 6 ) dogs received two intranodular injections of absolute ethanol (96%) via a through-the-endoscope injector, at weekly intervals; group B (n = 8) dogs were put on ivermectin (600 microg/kg BW, subcutaneously, twice, 14 days apart) and oral prednisolone (0.5mg/kg BW, every 12h, for a total of 3 weeks, tapering the dose accordingly). Clinical and fecal examination as well as endoscopy, were performed on admission and at 20, 60 and 180 days from the beginning of the treatment. One group A dog responded poorly and died of pyothorax during the trial and another developed esophagitis due to accidental intraluminal ethanol infusion, only to experience an uneventful recovery. At different times during the 6-month follow-up period, there was a complete disappearance of the clinical signs in 4/6 group A dogs. However, full nodular regression was achieved only in one dog, and parasitic ova were still found in the feces of 4/6 dogs. At the same period of time in five group B dogs still available for evaluation, resolution of the clinical signs and complete nodular regression were seen in four and five animals, respectively, while negative fecal results were obtained in all dogs (8/8) of the same group 2 months from the beginning of the treatment. No significant difference was found between the groups, regarding the resolution of clinical signs, though group B dogs demonstrated a significantly higher rate of regression of esophageal nodules as well as negative fecal results, compared to group A dogs. The combination of ivermectin and prednizolone may be considered an effective treatment in the symptomatic and evidently asymptomatic esophageal spirocercosis of the dog.

Spirocercosis-associated esophageal sarcomas in dogs. A retrospective study of 17 cases (1997-2003).

Seventeen client-owned dogs diagnosed with spirocercosis-associated esophageal sarcomas were retrospectively reviewed. The most common clinical signs noticed were vomiting and/or regurgitation (94%), lethargy and depression (59%), pyrexia and anorexia (41% each). Leukocytosis (82%) and microcytic hypochromic anemia (30%) were the most common hematological abnormalities. Caudal thoracic masses were demonstrated on survey radiographs of 13/15 of the dogs and thoracic spondylitis was detected in 12/15 dogs. Spirocerca lupi eggs were detected in 2/8 patients and worms were demonstrated on 1/11 at necropsy. Ten cases underwent surgical attempt to remove the tumors. In six of them partial esophagectomy (PE) was performed and all of them survived the immediate postoperative hospitalization. Five of the cases that underwent PE also received chemotherapy after surgery (doxorubicin (Adriamycin, Upjohn)) with an average survival time of 267 days. The histopathological results of the esophageal tumors were osteosarcoma (9), fibrosarcoma (5) and undifferentiated sarcoma (1). In areas endemic to spirocercosis, regurgitation or vomiting in dogs and microcytic hypochromic anemia and neutrophilia warrant ruling out esophageal sarcomas. Proper surgical treatment could prolong the dogs' lifespan for months, and improve their quality of life.

Molecular epidemiological survey on the vectors of Thelazia gulosa, Thelazia rhodesi and Thelazia skrjabini (Spirurida: Thelaziidae).

A Polymerase Chain Reaction (PCR)- based assay developed for the specific identification of Thelazia gulosa, Thelazia rhodesi and Thelazia skrjabini (Nematoda, Spirurida), which cause bovine ocular thelaziosis, was evaluated for its usefulness in detecting the intermediate hosts and in estimating the infection prevalence of vectors in field conditions throughout 5 years (from 1997 to 2001). A total of 5190 flies were captured and identified as Musca larvipara, Musca osiris, Musca autumnalis, Musca tempestiva or Musca domestica. Genomic DNA was extracted from pools constituted by heads, thoraces, abdomens and wings of 10 flies of each species, and 2076 samples were subjected to a PCR assay to specifically detect the ribosomal ITS-1 sequence of bovine Thelazia. Amplicons were sequenced and subjected to digestion with CpoI restriction enzyme. M. autumnalis, M. larvipara, M. osiris and M. domestica species were shown to be PCR positive. T. gulosa was specifically detected by PCR in M. autumnalis, M. larvipara, M. osiris and M. domestica, whereas T. rhodesi is in M. autumnalis and M. larvipara. Of 27 positive samples, 23 were positive for T. gulosa and 4 for T. rhodesi, with a mean prevalence of 2.86% in the whole fly population collected. The highest mean prevalence values of infection were detected in M. autumnalis (4.46%) and M. larvipara (3.21%), and the former species was confirmed to be the vector of T. gulosa and T. rhodesi. This study is the first report of M. osiris as a vector of T. gulosa and M. larvipara as a vector of T. gulosa and T. rhodesi under natural conditions. The occurrence of Thelazia in fly populations in the Apulia region of Italy (in the 5 grazing seasons considered) indicates that cattle thelaziosis is enzootic in southern Italy. This molecular assay should be a useful epidemiological tool for assessing the role of different species of flies as intermediate hosts of thelaziae.