Haematobium irritans and Haematobium titillans as potential vectors of Parabronema skrjabini in camels (Camelus bactrianus) in Inner Mongolia, China.

Parabronema skrjabini is one of the most harmful nematodes to camels and is responsible for economic losses in animal husbandry industry. There is an urgent need for in-depth studies of potential vectors of the nematode due to its scant regarding information. As previous studies indicated that flies may be the vectors of P. skrjabini, we captured flies in the main camel-producing areas of Inner Mongolia. After autopsy of the specimens of two species of horn flies, we observed the morphology of the suspected nematode larvae found in them. Internal transcribed spacer ribosomal-DNA gene sequences were considered the best candidate to confirm the species of the larvae found. Our results showed that the homology compared with P. skrjabini was 99.5% in GenBank. Subsequently, we preliminarily identified two species of horn flies through morphological observation and then sequenced the mitochondrial-DNA-gene cytochrome oxidase subunit I obtained from two species of horn flies, with 100 and 99.2% similarity to sequences deposited in GenBank, respectively. Thus, we identified Haematobia titillans and Haematobia irritans and provided evidence for their potential role as vectors of parabronemosis. Our study provides reference for future research on the life history of the nematode and the vectors of parabronemosis.

Rhabdochona angusticaudata sp. n. (Nematoda: Rhabdochonidae) from the Japanese eel Anguilla japonica, and new records of some other nematodes from inland fishes in Japan.

Collections of parasitic nematodes of fishes from some freshwater and brackish-water localities in Honshu, Shikoku and Hokkaido, Japan, carried out in 1996-2009, yielded the following eight species: Hysterothylacium haze (Machida, Takahashi et Masuuchi, 1978) (Anisakidae), Paraquimperia sp., Pingus sinensis Hsü, 1933 (both Quimperiidae), Heliconema anguillae Yamaguti, 1935 (Physalopteridae), Rhabdochona angusticaudata sp. n., Rhabdochona zacconis Yamaguti, 1935 (both Rhabdochonidae), Ascarophis arctica Polyanskiy, 1952 (Cystidicolidae) and Eustrongylides sp. larvae (Dioctophymatidae). The new species, R. angusticaudata sp. n., described from the intestine of the Japanese eel Anguilla japonica Temminck et Schlegel in the Renjoji River, Shikoku, is mainly characterised by the presence of 14-16 anterior prostomal teeth, non-bifurcated deirids with rounded ends, the length of the left spicule (300-384 µm long), the right spicule with a dorsal barb and non-filamented eggs. The species of Paraquimperia Baylis, 1934 and A. arctica are reported from Japan for the first time. Based on light and scanning electron microscopical examinations, some new morphological data on H. haze, H. anguillae, R. zacconis and A. arctica are provided.

The larval development of Habronema muscae (Nematoda: Habronematidae) affects its intermediate host, Musca domestica (Diptera: Muscidae).

Although the life cycle of the equid stomach parasite Habronema muscae was disclosed more than 100 years ago, little is known about the effect of the developing nematode larvae in its intermediate host, Musca domestica. In a series of experiments, freshly hatched M. domestica larvae were exposed to H. muscae eggs contained in a faecal sample of a naturally infected horse. In daily intervals, 50 fly larvae were removed and transferred on a parasite-free larval rearing medium where they completed their development. Hatched flies were examined for the presence of Habronema third-stage larvae. In two subsequent control groups, flies spend their entire larval life in contaminated horse faeces and in a parasite-free larval rearing medium, respectively. Out of the 700 fly larvae used in the infection experiments, 304 developed into adult flies of which 281 were infected. The average nematode larval burden rose from 3.6 in the group with the shortest exposure to more than 25 in the groups with the longest exposure. The proportion of larvae that developed into the adult insect fell from 82 % in the uninfected control group to 27 % in the positive control group. The pupae of the positive control group were smaller and lighter than those of the uninfected control group. Lower pupal size and weight in the positive control group as well as a lower insect developing rate might be attributed to the destruction of adipose cells in the maggots by Habronema larvae.

Light and scanning electron microscopical examination of the third stage larva of Physocephalus dromedarii (Nematoda: Spirocercidae)--an abomasal nematode of the one humped camel (Camelus dromedarius).

The life cycle of Physocephalus dromedarii was studied under experimental conditions. Larvae obtained from naturally infected Scarabaeus cristatus and Aphodius sp. (Coleoptera: Scarabaeidae) were measured and examined in light and scanning electron microscopy and used to infect a dromedary as final host as well as chicken, mice and a toad as possible paratenic hosts. Larvae with the same morphology and similar measurements were found in naturally infected reptiles (Trapelus flavimaculatus, Eryx jayakari, Cerastes gasperettii). Body length of examined larvae varied between 1450 and 1700 µm. Dorsal, ventral and lateral lips, peg-like papillae and amphideal pits are located on the cephalic cone. There are two asymmetrical cervical deirids, long simple lateral wings and a knob-like posterior end covered with minute spines. In the camel, patency is reached within 12 weeks after infection while larvae in paratenic hosts migrate into the wall of the alimentary tract and become dormant.

Female host sex-biased parasitism with the rodent stomach nematode Mastophorus muris in wild bank voles (Myodes glareolus).

Abundance and prevalence of helminth infections often differ between host sexes, and are usually biased in favor of males. Relatively few cases of female-biased parasitism have been reported. We sampled bank voles in three woodland sites in N.E. Poland over 11 years at 3-4-year intervals, and assessed their parasite burdens. Prevalence and abundance of the stomach nematode Mastophorus muris were consistently higher among females. Among adult female bank voles from the two sites that showed the highest prevalence with M. muris, both prevalence and abundance were significantly higher in lactating bank voles, but not pregnant animals, and the effect of lactation was evident in both sites, in all four surveys, and in both age classes. Although the magnitude of the effect of lactation varied between years, it was not confounded by any significant interactions with other factors. We hypothesize that mature and reproductively active female bank voles are subject to higher exposure compared with males of similar age, as a consequence of the increased content of invertebrates in their diet, including the intermediate hosts of M. muris, required to meet the higher increased energy and protein demands of nursing litters throughout the summer months.

Behavioural changes in the flour beetle Tribolium confusum infected with the spirurid nematode Protospirura muricola.

We examined changes to the behaviour of flour beetles, Tribolium confusum, infected with the rodent stomach worm, the spirurid Protospirura muricola, in the context of the 'Behavioural Manipulation Hypothesis'. Trobolium confusum infected with the third-stage infective larvae of P. muricola showed consistently altered patterns of behaviour. Relative to uninfected beetles, over a measured time period, beetles infected with P. muricola were likely to move over a shorter distance, when moving their speed of movement was slower, they were more likely to stay in the illuminated area of their environment, more likely to emerge from darkened areas into the illuminated areas, and their longevity was significantly shortened. The changes in behaviour, as reflected in effects on speed of movement, were only evident among beetles that actually harboured infective cysts and not among those carrying younger infections when the larvae within their haemocoels would have been at an earlier stage of development and not yet capable of infecting the definitive murine hosts. We discuss whether these changes would have made the beetles more susceptible to predation by rodents, and specifically by the omnivorous eastern spiny mouse, Acomys dimidiatus, the natural definitive host of this parasite in Egypt, from where the P. muricola isolate originated, and whether they support the Behavioural Manipulation Hypothesis or reflect parasite-induced pathology.

Habronema muscae (Nematoda: Habronematidae) larvae: developmental stages, migration route and morphological changes in Musca domestica (Diptera: Muscidae).

The present paper describes the morphological modifications occurring during the larval development of Habronema muscae (Nematoda: Habronematidae) in Musca domestica (Diptera: Muscidae), along with the reactions caused by parasitism and the migration route of the nematodes inside the flies. Houseflies were reared on faeces of a H. muscae-infected horse, then dissected and processed by histology. The experimental part of the study was performed in 1996 in the Parasitological Experimental Station W.O. Neitz, Federal Rural University of Rio de Janeiro, Seropédica, Rio de Janeiro State, Brazil. Three different larval stages of H. muscae were recovered, measured and described. The encapsulation of larval nematodes was found in the third larval instar (L3) of M. domestica and cryptocephalic pupa. The mature capsules were observed in dipteran L3, pupae and mainly adults. In 1day-old or more M. domestica adults an active rupturing of capsules by H. muscae L3 and the migration to the head through the circulatory system and insect hemocoel were observed. Infective H. muscae L3s remained exclusively in the head of adult 5days-old or more M. domestica.

Can a specialist parasite species of a widespread and common host species be rare? The case of Spinitectus inermis (Nematoda: Cystidicolidae) in eels Anguilla anguilla.

The claim by many authors that Spinitectus inermis (Zeder, 1800), a narrowly specific parasite of European eels Anguilla anguilla (L.), is a rare species is considered at three levels: its geographical range, its frequency of occurrence compared to other eel parasites and its relative abundance in component communities. The parasite is widely distributed in freshwater throughout the European range of the eel but its occurrence is erratic and unpredictable, being known from only 8 countries. Surveys of eel parasites in the United Kingdom and in Continental Europe show that it is present in only 13% of British and 29% of continental localities. This satisfies one of the criteria for rarity. When present, its prevalence ranges from 1.8% to 43.3%, so it can be considered rare in some localities but in a few it may be common and on occasion it may be the dominant species in the gastro-intestinal community. Populations of S. inermis are almost always characterised by high levels of overdispersion, even at low prevalence. The species also displays an ability to colonise a locality following introduction there. Overall it meets many of the criteria of a rare species including a restricted distribution and a low frequency of occurrence and so it can be considered to exhibit diffusive rarity.

[A new nematode species, Physaloptera musayevi sp. n., from the Lybian jird Meriones libycus and the social vole Microtus socialis in the Gobustan area of Azerbaijan Republic].

Nematodes belonging to the genus Physaloptera Rudolphi, 1819 were obtained from the Lybian jird Meriones libycus and the social vole Microtus socialis in the Gobustan area as a result of helminthological research. These nematodes significantly differed from other species of the genus in morphological and morphometric characters. In the present paper, they are described as the new species Physaloptera musayevi sp. n.

Identification of the intermediate host of Gongylonema sp., the etiological agent of the necrotic oropharyngeal disease of the Scops owl (Otus scops).

Since 1997, fledgling Scops owls (Chordata: Strigidae) have been brought to the Brinzal Owl Rescue Centre (Madrid, Spain) with severe lesions in their oral cavities. Lesions consist of the presence of proliferative necrotic material in the oral cavity resulting in white plaques, which can lead to death by starvation. This disease has been detected in owls only within the limits of the city of Madrid. The etiologic agent has been identified as Gongylonema sp. (Nematoda: Spirurida), a nematode genus that includes a coprophagous arthropod as intermediate host in its cycle. The aim of this study was to identify the intermediate host of the parasite. Our work was structured in four component phases: i) Diet study of newborn chicks; ii) trapping arthropods that could be intermediate hosts; iii) molecular detection of the parasite in the selected arthropods: and iv) molecular characterization of the detected parasites by amplifying the cox1 gene. Four male owls were radio-tagged in order to locate their nests and a camera trap was placed to identify the prey brought to the owlets. Secondly, the arthropods present in the hunting areas of the owls were sampled, identified and analyzed by real time PCR (rtPCR). Only oriental cockroaches, B. orientalis (Arthropoda: Blattodea), were positive by rtPCR detection of Gongylonema sp. (66.7%). The nematodes obtained from cockroaches had a 99.8% identity of the cox1 gene with the Gongylonema sp. isolated for the first time in a Scops owl. Furthermore, these sequences only showed an <89% identity with all the other Gongylonema sequences available in the GenBank database. We conclude that the oriental cockroach should be considered as an intermediate host of the etiologic agent of NOD.

A survey of dung beetles infected with larval nematodes with particular note on Copris lunaris beetles as a vector for Gongylonema sp. in Iran.

Dung beetles (family Scarabaeidae) are one of the largest families of beetles worldwide. Due to biological behavior of these arthropods, they are considered to play an important role in the life cycle of some helminths. In the present study, dung beetles collected from cattle pastures in rural areas of Ardabil province, north-west of Iran were examined for infection with larval stages of helminths. According to the results, nematodes of 2 genera were identified including Rhabditis and Gongylonema. The more common species was Rhabditis sp. which was found in 9 species of beetles. Out of 15 different species of dung beetles, Copris lunaris was the only scarabaeid to be found naturally infected with the larval stages of Gongylonema sp. Our new findings introduce C. lunaris as a potential biological vector for transmission of Gongylonema sp. to vertebrates in the surveyed region.

Identification of the intermediate hosts of Habronema microstoma and Habronema muscae under field conditions.

A polymerase chain reaction (PCR)-based assay was used for the specific detection of Habronema microstoma and Habronema muscae (Nematoda, Spirurida) in order to identify the intermediate hosts of both nematode species under field conditions. A total of 1087 netted and 165 laboratory-bred flies were tested. Flies were identified as Musca domestica Linnaeus 1758, Musca autumnalis De Geer 1776, Haematobia irritans (Linnaeus 1758), Haematobia titillans (De Geer 1907) and Stomoxys calcitrans (Linnaeus 1758) (Muscidae). Genomic DNA was extracted from pools of fly heads, thoraces and abdomens, and 703 samples were subjected to a duplex two-step semi-nested PCR assay to specifically detect diagnostic regions within the ribosomal ITS2 sequence of both H. microstoma and H. muscae. Stomoxys calcitrans specimens were positive for H. microstoma DNA and M. domestica specimens were positive for H. muscae DNA. In particular, PCR-positive samples derived from both farm-netted and laboratory-bred flies. The present study represents the first evidence of the vectorial competence of different fly species as intermediate hosts of Habronema stomachworms under field conditions. We discuss the roles of S. calcitrans and M. domestica in transmitting H. microstoma and H. muscae.

Acute phase proteins, interleukin-6, tumor necrosis factor, nitric oxide and oxidative stress markers in horses with cutaneous habronemosis under field condition.

Habronemosis is a common parasitic disease of horses worldwide. In order to investigate how haptoglobin (Hp), serum amyloid A (SAA), oxidative stress markers, nitric oxide (NO), interleukin-6 (IL-6) and tumor necrosis factor (TNF- α), varies in cutaneous habronemosis, 30 horses with the clinical disease and 20 clinically healthy horses were included in the current study. The serum levels of Hp, SAA, and proinflammatory cytokines (IL-6 and TNF-α), NO, malondialdehyde (MDA), super oxide dismutase (SOD), glutathione (GSH), and total antioxidant capacity (TAC) were determined in horses before and after two weeks of treatment. The serum levels of Hp, SAA, IL-6, TNF-α and MDA were significantly elevated in infected horses as compared to the controls. Alternately, the serum levels of SOD, GSH, TAC and NO, were recorded low in infected horses as compared to the controls. All tested markers resumed the same levels after treatment as in control group. The Hp, SAA, IL-6, TNF-α, and MDA exhibited a high degree of clinical accuracy of the cases diagnosis. The area under the curve (AUC) for acute phase proteins (SAA, Hp), IL-6, TNF-α, and MDA was 0.87, 0.94, 0.96, 0.96 and 1.0, respectively. These findings showed that Hp, SAA, IL-6, TNF-α, and MDA may be supportive in the diagnosis of cutaneous habronemosis in horses and, simultaneously, they can also be used to monitor the progress of the treatment.

A review of Gongylonema spp. (Nematoda: Gongylonematidae) in North American rodents with description of a new species from the cotton rat, Sigmodon hispidus (Mammalia: Cricetidae).

Gongylonema archboldi n. sp. (Nematoda: Gongylonematidae) is described from tunnels in the gastric mucosa of the stomach of the cotton rat (Sigmodon hispidus) from Highlands County, Florida, U.S.A. Measurements are also given for specimens from cotton mice (Peromyscus gossypinus), oldfield mice (Peromyscus polionotus), Florida mice (Podomys floridanus), and golden mice (Ochrotomys nuttalli) from the same locality. Additional specimens were collected from the cotton rat and the rice rat (Oryzomys palustris) from Berry Island, San Patricio County, Texas. The new species is differentiated from congeners by a combination of the following characters: length of the left spicule, length and shape of the gubernaculum, distribution of cuticular bosses, length of esophagus, and distance of the vulva from the posterior end. The status of the genus Gongylonema in North American rodents is reviewed.

Further observations on the development of Gongylonema pulchrum in rabbits.

Third-stage larvae of Gongylonema pulchrum from naturally infected dung beetles were inoculated orally into 24 rabbits. Worm recovery ranged from 54 to 91% (mean = 67.5%) during the period from 24 hr to 52 wk postinoculation (PI). Two hours PI, the larvae entered the mucosa at the junction of the stomach and esophagus and migrated upward. Early development occurred primarily in pharyngeal mucosa, tongue, and buccal mucosa. The third molt took place 11 days PI and the final molt at 36 days PI. Male worms reached sexual maturity at 7 wk PI and females at 9 wk PI. Adult worms were found mainly in the esophagus but also occurred in the tongue and the wall of the oral cavity after 30 wk PI. Embryonated eggs appeared in the feces of 3 rabbits inoculated with 50 or 100 larvae on days 72-81 PI. Morphologically, the cuticle in young fourth-stage larvae exhibited bosses on the anterior portion on day 11 PI, and the left spicule length : total body length exhibited no remarkable change between 9 and 52 wk PI. The latter finding confirms the utility of the ratio for identification of the nematode.

The influence of food competition and host specificity on the transmission of Triaenophorus crassus (Cestoda) and Cystidicola farionis (Nematoda) to Coregonus lavaretus and Coregonus albula (Pisces:Coregonidae) in Finland.

As a superior competitor for planktonic food, vendace (Coregonus albula), when abundant, is expected to displace whitefish (Coregonus lavaretus) from feeding on plankton and to force it to rely more on benthic food. The predicted result would be a reduced abundance of the copepod-transmitted cestode Triaenophorus crassus in whitefish, but an increase in the abundance of the nematode Cystidicola farionis transmitted via benthic amphipods. We studied the occurrence of both parasites in whitefish during 1991-1996 in three interconnected areas at Lake Saimaa, Finland, where the densities of the vendace stocks varied due to natural fluctuation in year-class strengths. In accordance with our hypothesis, some indication of the effect of the density of the vendace population on abundance of C. farionis infection in whitefish was found, but not in the case of T. crassus. Only 0.2% of vendace were infected with T. crassus, while up to 100% of the whitefish in the yearly samples harboured the parasite. In further experiments we clarified which copepod species in Lake Saimaa act as first intermediate hosts of T. crassus, and verified from stomach samples the exposure of both whitefish and vendace to those species. Experimental infections indicated that the infectivity of T. crassus is lower for vendace than for whitefish. We suggest that the reason why vendace stock density does not affect T. crassus infection in whitefish is that T. crassus is transmitted in littoral areas during a short period in spring. At that time of the year copepods are abundant and available to both whitefish and vendace, but since the parasite is less infective to vendace. they do not become infected. At other times of the year, dense vendace stocks may force whitefish to shift to benthic food, which includes amphipods transmitting C. farionis. During vendace stock decline, whitefish may, however, continue to feed on plankton and avoid exposure to C. farionis.

Tetrameres americana Cram (1927) populations in chickens infected with different dose levels.

Three groups of 20-45 Lohman brown chickens aged 3 weeks were orally infected with doses of 25, 100 and 400 Tetrameres americana, respectively. Fifteen chickens were kept as uninfected controls. Every third week in a period of 12 weeks, 5-15 chickens were slaughtered and the proventriculi were examined for the presence of adult stages of T. americana. From day 21 post-infection, pooled feacal samples were examined for parasite eggs, whereas the weight gain of the chickens was monitored weekly. The parasite established the infection in similar rates in all the three groups, 9.5-15.2%, except on day 84 post-infection, when the establishment rate of the high-dose group was significantly lower (P < 0.005). The average worm burden increased with increasing dosages, though displaying the worm burden of the high-dose group as being roughly halved on day 84 post-infection, thus suggesting an expulsion of worms. Females were more abundant than males. The mean lengths of male and female specimens showed no significant differences between the groups. The egg output was also increased with increasing dosages with the earliest prepatent period of 38 days post-infection found in the high-dose group. Infected chickens exhibited no difference in weight gain in comparison with the controls. This study demonstrated that single infections with varying doses influenced the establishment rate and the worm burden but not the parasites egg excretion, worm size nor the weight gain and mortality of the final host.

Pathology associated with physalopterid larvae (Nematoda: Spirurida) in the gastric tissues of Australian reptiles.

Cysts containing one or more physalopterid larvae were found commonly in the gastric tissues of snakes (Elapidae) and in all five families of lizards (Agamidae, Gekkonidae, Pygopodidae, Scincidae and Varanidae) in Western Australia. Snakes had been collected from many locations in tropical and arid Western Australia between 1912 and 1976, and lizards from the Great Victoria Desert between September 1978 and March 1979. Most cysts occurred in the submucosa; others were found within stomach muscle, and subserosally on the external stomach surface. Encysted and apparently viable larvae were associated with minimal host inflammatory response. Encysted dead and degenerating larvae occurred in cysts with inflammatory cell infiltrates, principally in snakes.

Migration of a spiruroid nematode through the brain of a horse.

A pregnant 10-year-old Paint mare was examined because of an acute neurologic disturbance. Physical examination revealed signs consistent with extensive, asymmetric brain stem disease. The hemogram, serum chemical panel, and results of lumbosacral spinal fluid analysis were within normal limits. A primary diagnosis of equine protozoal myeloencephalitis was considered, and the mare was placed on treatment with trimethoprim-sulfadiazine. After 5 weeks of steady improvement, an acute exacerbation of neurologic signs necessitated euthanasia of the mare. At necropsy, large, malacic tracts were found extending through the brain stem and cerebral cortex. Cross sections of a nematode were observed microscopically and subsequently were identified as belonging to a single gravid female Draschia megastoma.

On Physaloptera sibirica Petrow et Gorbunow, 1931 (Nematoda: Physalopteridae) parasitizing Iberian mammals.

Faunistic and morphological study of Physaloptera sibirica PETROW et GORBUNOW, 1931 (Nematoda: Physalopteridae), a nematode isolated in Spain from three mammal species: red fox, Vulpes vulpes (LINNAEUS, 1758) (Carnivora: Canidae), eurasian badger, Meles meles (LINNAEUS, 1758) (Carnivora: Mustelidae), and garden dormouse, Eliomys quercinus (LINNAEUS, 1766) (Rodentia: Myoxidae) in the Iberian Peninsula. Morphological features of cephalic and caudal regions of males are presented for the first time by SEM (scanning electron microscopy).