Eye trematode infection in small passerines in Peru caused by Philophthalmus lucipetus,an agent with a zoonotic potential spread by an invasive freshwater snail

Until now, four species of eye trematodes have been found in South America. Of them, Philophthalmus lucipetus (synonymized with Philophthalmus gralli) displays a broad host spectrum, with at least 30 bird species (prevalently large water birds), five mammal species and humans serving as definitive hosts, and with snails Fagotia (Microcolpia) acicularis, Amphimelania holandri, Melanopsis praemorsa and Melanoides tuberculata serving as intermediate hosts. When examining a total of 50 birds of ten species in the wetland of Pantanos de Villa, Lima, Peru in July 2011, eye trematodes were identified visually in the edematous conjunctival sac of 11 (48%) out of 23 resident many-colored rush tyrants Tachuris rubrigastra. Based on morphometric characteristics, the trematodes were identified as P. lucipetus. ITS2 and CO1 gene of the examined specimens combined showed a 99% similarity to an Iranian isolate of Philophthalmus sp. from the intermediate host Melanoides tuberculata, an invasive freshwater snail, suggesting that these two isolates represent the same species with a wide geographical range. Moreover, the prevalence of infection with the philophthalmid cercariae was 31% in 744 Melanoides tuberculata examined in Pantanos de Villa in 2010. It is evident that P. lucipetus occurs throughout the world as well as locally, including Eurasia and South America. Here we report this trematode for the first time in Peru, and we were the first to sequence any of the South American eye trematodes. Low host specificity of P. lucipetus and the invasive character of Melanoides tuberculata as a competent intermediate host suggest that eye trematodosis caused by P. lucipetus may emerge frequently in various parts of the world, especially in the tropics. Increase of the zoonotic potential of the P. lucipetus associated with this invasive snail spreading across the world is predictable and should be of interest for further research.     

Lassen sich Silber-, Wei?kopf- und Heringsm?we (Larus argentatus, L. cachinnans, L. fuscus) molekulargenetisch unterscheiden?

Zusammenfassung Ein Teilstück des mitochondriellen Cytochrom-b Gens vonLarus argentatus, L. cachinnans michahellis undL. fuscus wurde mittels PCR amplifiziert und sequenziert. Der Sequenzvergleich mittels PAUP 3.1 zeigt, daß Weißkopfmöwe und Silbermöwe genetisch klar getrennt sind und sich durch 2% Basenaustausche unterscheiden. Weißkopf- und Heringsmöwe stehen sich näher als Weißkopf- und Silbermöwe. Diese 3 Möwenarten unterliegen keinem signifikanten Genfluß und bilden vermutlich eine Superspezies. Aus der Dimension der Basenaustausche kann man schätzen, daß es sich um relativ junge, vielleicht 100 000–500 000 Jahre alte Taxa handeln muß. Die Abtrennung vonL. cachinnans als eigene Art ist molekulargenetisch demnach gerechtfertigt.

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Genetic distinction ofLarus argentatus, L. fuscus andL. cachinnans

The mitochondrial cytochrome b gene ofLarus argentatus, L. fuscus andL. cachinnans michahellis was amplified by PCR and partially sequenced. Sequence analysis by PAUP 3.1 indicated thatL. argentatus andL. cachinnans are genetically distinct and separated by 2% base exchanges.L. cachinnans seems to be closer related toL. fuscus than toL. argentatus. The three taxa represent a superspecies. Our data imply that gene flow between the 3 gull species must be negligible and furthermore, that they represent rather young species which may have split 100 000–500 000 years ago. The recognition ofL. cachinnans as a distinct species is supported by molecular evidence.

     

Molecular characterization of Oxyspirura mansoni and Philophthalmus gralli collected from the eyes of domestic chickens in Bangladesh

A variety of helminths have been found in domestic chickens in Bangladesh, but little is known about their gene sequences. Here, parasitic nematodes and trematodes were collected from the eyes of domestic chickens and analyzed for their morphological and morphometric characteristics, and characterized molecularly. The helminths were identified as Oxyspirura mansoni and Philophthalmus gralli. The ITS1 and ITS2 sequences of O. mansoni were 532 bp and 306 bp in length, respectively, and showed low identity (50.7–62.7%) with those of O. petrowi and O. conjunctivalis. Furthermore, the O. mansoni CO1 sequences (393 bp) showed five haplotypes (97.5–99.5% similarity) that formed a monophyletic clade. With respect to P. gralli, the ITS1 (452 bp) and ITS2 (736 bp) sequences showed 100% similarity with the reference sequences in GenBank. Both the ND1 and CO1 phylograms showed that P. gralli from Bangladesh, Costa Rica and Peru form a monophyletic clade, distinct from the clades of P. lucipetus and P. lacrymosus. Our data show that, Philophthalmus gralli isolates from Bangladesh, Costa Rica and Peru are genetically close to each other.     

Impacts of an endoparasitic copepod, Ismaila belciki, on the reproduction,growth and survivorship of its nudibranch host, Janolus fuscus

Copepods from the genus Ismaila are large endoparasites that inhabit the main body cavity and/or cerata of opisthobranch molluscs. These parasites exhibit many life history characteristics typically found in parasitic castrators, yet the actual impact of infection on reproduction, growth or survivorship of the hosts are unknown. On the Oregon (USA) coast, Ismaila belciki can infect over 80% of their hermaphroditic hosts, Janolus fuscus. In laboratory mating experiments, we compared the reproductive output (egg mass weight, number of egg capsules, number of viable embryos) and the gonadal somatic index of infected versus uninfected J. fuscus. Infected J. fuscus could produce viable sperm and copulate. Mating with an infected individual did not limit a sea slug’s reproductive output. However, infected J. fuscus had significantly lower reproductive output (by 34–54%), producing smaller egg masses with fewer capsules and viable embryos. Infected hosts had significantly lower gonadal somatic index than their uninfected counterparts, although there was no significant difference in gonadal somatic index between hosts with single and double infections. By collecting the egg sacs produced by the copepod parasite during experiments, we estimated that 25–34% of the host’s reproductive output is usurped by the parasite and re-directed to the parasite’s own reproduction. In the laboratory, infection did not alter growth in J. fuscus. However, infection significantly decreased survivorship in mature (but not immature) nudibranch hosts. These results suggest that I. belciki is not a true castrator, but it does reduce the reproductive output of its host and may therefore limit the natural population size of J. fuscus.     

Focal Philophthalmus gralli infection possibly persists in Melanoides tuberculata over two years following the definitive hosts' removal

Philophthalmosis is a zoonotic disease associated largely with the spread of the invasive freshwater snail Melanoides tuberculata, serving as an intermediate host. Here we examined Philophthalmus gralli focal fenced infection site reported recently as being associated with Tinamus major and M. tuberculata in Alajuela, Costa Rica. Removal of the definitive hosts allowed us to address also the long-term survival strategy of the parasite. Initially, the snail intermediate hosts displayed high prevalence of P. gralli infection across all its age cohorts. Two years following the removal of definitive hosts, the infection rate decreased by one order of magnitude, while the snails aging less than one year displayed zero infection prevalence. Additionally, phylogenetic analysis of mitochondrial (ND1) and nuclear (ITS1, ITS2) DNA loci revealed negligible intrasite DNA variability of the specimens obtained at the study site in Costa Rica (but not of those obtained earlier in Peru or New Zealand), supporting strongly the hypothesis on focal origin of the infection. The observed dynamics of infection suggests the explanation for the high variability in P. gralli prevalence in intermediate hosts experienced worldwide. We noticed that the reports claiming > 20% prevalence of M. tuberculata infection by P. gralli originated exclusively from foci with known eye infection of the definitive hosts, while the P. gralli infection penetrance < 2% is typically associated with sites, where the infection of definitive hosts was not observed, suggesting that the infected definitive hosts were present onsite only in the past, or were present only at a site upstream or downstream of the respective sampling site. Thus, this is the first evidence on the possible persistence of eye-trematode infection site for over two years following the last confirmed outbreak in its adult hosts.     

Mating behavior of Philophthalmus megalurus and P. gralli in concurrent infections of chicks

Nollen P. M. 1984. Mating behavior of Philophthalmus megalurus and P. gralli in concurrent infections of chicks. International Journal for Parasitology14: 71–74. Mating patterns were determined in concurrent infections of the eyeflukes of birds, Philophthalmus megalurus and P. gralli, by detection of ³H-tyrosine-labeled sperm on autoradiograms. When labeled P. megalurus adults were transplanted singly with unlabeled P. gralli adults, interspecies mating took place. When both unlabeled P. gralli and P. megalurus were transplanted with single labeled P. megalurus, sperm were transferred to both species. However, the insemination rate of P. gralli was much lower than when unlabeled P. meglaurus adults were absent. Labeled P. gralli transplanted singly with unlabeled P. megalurus or unlabeled adults of both species did not show interspecies mating, but carried on intraspecies mating at a normal rate. A low rate of self-insemination was observed in labeled worms when the opposite species was present, but not when the same species was present. Evidence for interspecies fertilization and hybridization between P. megalurus and P. gralli was not found.     

The natural occurrence of Pandora heteropterae (Zygomycetes: Entomophthorales) infecting Lygus lineolaris (Hemiptera: Miridae)

An unknown fungal pathogen was recovered from Lygus lineolaris (Palisot de Beauvois) during a survey of parasitic and pathogenic natural enemies conducted in Franklin County, Arkansas. The pathogen was identified as Pandora heteropterae (Ba?azy) Keller based on characteristics of the morphology, as well as growth and sporulation on hosts. The fungus infected 11 of the 3405 (0.32%) wild L. lineolaris collected. In a laboratory host-range bioassay, five of seven hemipteran species from the families Miridae, Coreidae, Lygaeidae, and Pentatomidae were successfully infected. P. heteropterae was previously reported only once, from an unidentified host species in Poland. Here we describe the morphology and growth of P. heteropterae and discuss its potential impact on L. lineolaris in the field.     

Rapid long‐distance migration in Norwegian Lesser Black‐backed Gulls Larus fuscus fuscus along their eastern flyway

We studied the long‐distance migration of Lesser Black‐backed Gulls Larus fuscus fuscus breeding in northern Norway along their eastern flyway using geolocators in 2009 and 2010. The majority of birds wintered in lakes in East Africa and the southeast Mediterranean was the most important stopover area. Larus f. fuscus along the eastern flyway travelled at a net travel speed of 399 and 177 km/day during the autumn and spring migration, respectively, higher than published travel speeds for Dutch Larus fuscus migrating along the western flyway. The results suggest that the long‐distance migratory Norwegian L. f. fuscus seek to minimize time spent in transit, whereas lower travel speed during northerly spring migration may reflect differences in wind patterns or food conditions between spring and autumn.     

A physiological marker for quantifying differential reproductive investment between the sexes in Yellow-legged gulls (Larus michahellis)

Asymmetry between males and females in the energy they invest initially in reproduction has resulted in the evolution of differing reproductive strategies (caring females vs. competitive males). However, parental care in many birds is shared by both sexes suggesting that male energy expenditure in agonistic behaviors and courtship feeding might compensate female costs of clutch production. Here, we tested the hypothesis that initial investment in reproduction by both sexes in the Yellow-legged Gull (Larus michahellis), a species with biparental care, is similar from a physiological perspective. In this income breeder, female and male reproductive investment during early breeding can be ultimately related to muscular activity (local foraging effort required for clutch production in females and courtship feeding and agonistic behaviors in the case of males). Thus, we evaluated sex-specific patterns of creatine kinase (CK, IU/L) levels in plasma, an indicator of physical effort associated with muscular activity dependent behaviors, through incubation as a reflection of the physiological response of both sexes to the reproductive investment they made up to clutch completion. Raw levels of CK were related to plasma levels of total proteins (TP, g/dL) to account for the differential physiological state of individuals when sampled (i.e. differential dehydratation). Female costs of clutch production were associated with post-laying levels of CK/TP. We grouped females according to their relative investment in clutch production: < 15.8%, 15.8 to 17.3% and > 17.3% of field metabolic rate; which showed increasing values of CK/TP (24.6, 53.1 and 66.0 IU/g, respectively). Moreover, we found similar CK/TP trends throughout incubation for both sexes (CK/TP = 50.2− [3.3 × days from laying]) suggesting similar physiological responses to reproductive effort and, therefore, analogous sex-specific initial investment. Thus, male investment in agonistic behaviors and courtship feeding apparently equaled female investment in clutch production. The use of CK measurements is revealed as a useful approach to investigating overall investment in reproduction for both sexes, providing further insights into our comprehension of reproductive strategies in seabirds.     

Sleep in the herring gull (Larus argentatus)

Sleep postures and eye state of free-ranging herring gulls (Larus argentatus) were studied during the breeding season. Three mutually exclusive behaviours were observed, namely sleep, rest-sleep and rest postures. Arousal thresholds, eye blink rates and eye closure time were obtained during these behaviours. Significant relationships existed between eye blinking, eye closure, and a raised threshold of arousal when birds were in the sleep and rest-sleep postures. During a natural disturbance, birds in the sleep posture remained in this posture but did not blink their eyes: this is called pseudo sleep. Male gulls also exhibited a lower threshold of arousal while in the sleep posture compared with females. We conclude that rhythmic eye blinking is a good indication of sleep in herring gulls.     

Effects of a novel entomopathogenic nematode-infected host formulation on cadaver integrity, nematode yield, and suppression of Diaprepes abbreviatus and Aethina tumida

An alternative approach to applying entomopathogenic nematodes entails the distribution of nematodes in their infected insect hosts. Protection of the infected host from rupturing, and improving ease of handling, may be necessary to facilitate application. In this study our objective was to test the potential of a new method of formulating the infected hosts, i.e., enclosing the infected host in masking tape. Tenebrio molitor L. cadavers infected with Heterorhabditis indica Poinar, Karunakar and David or Steinernema carpocapsae (Weiser) were wrapped in tape using an automatic packaging machine; the machine was developed to reduce labor and to standardize the final product. The effects of the tape formulation on the ability to protect the cadavers from mechanical damage, nematode yield, and pest control efficacy were tested. After exposure to mechanical agitation at 7-d-post-infection, S. carpocapsae cadavers in tape were more resistant to rupture than cadavers without tape, yet H. indica cadavers 7-d-post-infection were not affected by mechanical agitation (with or without tape), nor was either nematode affected when 4-d-old cadavers were tested. Experiments indicated that infective juvenile yield was not affected by the tape formulation. Laboratory experiments were conducted measuring survival of the root weevil, Diaprepes abbreviatus (L.), or the small hive beetle, Aethina tumida Murray, after the application of two H. indica-infected hosts with or without tape per 15 cm pot (filled with soil). A greenhouse experiment was also conducted in a similar manner measuring survival of D. abbreviatus. In all experiments, both the tape and no-tape treatments caused significant reductions in insect survival relative to the control, and no differences were detected between the nematode treatments. Fifteen days post-application, the infected host treatments caused up to 78% control in A. tumida, 91% control in D. abbreviatus in the lab, and 75% in the greenhouse. These results indicate potential for using the tape-formulation approach for applying nematode infected hosts.     

Development, growth, and migration of Philophthalmus gralli (Trematoda) from Jordan in the eyes of chicks

The growth of Philophthalmus gralli from Jordan was studied experimentally by inoculating 10 excysted metacercariae around the right eye of 47 day-old chicks. The growth rate was slow in the first three days during which flukes did not exceed 0.30 mm long. Then growth became rapid reaching 2.70 mm long on day 24. This was followed by a low growth rate during which the flukes grew from 2.70 mm to 2.90 mm on day 40. The flukes started to appear in the left eye on day 12 post-inoculation of metacercariae in the right eye. The recovery of flukes in the left eye increased from 5% on day 12 to 30% on day 32. The development of P. gralli from Jordan was the same as of other species of Philophthalmus. However, a sixth stage, post-maturation stage, is suggested in addition to the 5 developmental stages of Philophthalmus. The ratio of the transverse diameter of the ovary to that of testes in the post-maturation stage was 1:1.1-1:1.2.     

Litomosoides sigmodontis: A simple method to infect mice with L3 larvae obtained from the pleural space of recently infected jirds (Meriones unguiculatus)

Litomosoides sigmodontis is a filarial nematode that is used as a mouse model for human filarial infections. The life cycle of L. sigmodontis comprises rodents as definitive hosts and tropical rat mites as alternate hosts. Here, we describe a method of infecting mice with third stage larvae (L3) extracted from the pleural space of recently infected jirds (Meriones unguiculatus). This method enables infection of mice with a known number of L3 larvae without the time-consuming dissection of L3 larvae from mites and results in higher worm recovery and patency rates than conventional methods. Additionally, this method allows for geographical separation of the facility maintaining the L. sigmodontis life cycle from the institution at which mice are infected.     

Pathology and first report of natural eye infection with the trematode Philophthalmus gralli (Digenea,Philophthalmidae) in Tinamus major (Tinamiformes,Tinamidae), Costa Rica

The eye-fluke Philophthalmus gralli (Philophthalmidae Looss, 1899) was found in six birds known as great tinamous (Tinamus major) reared in a wild animal shelter located in Alajuela, Costa Rica. The birds presented conjunctival hyperemia, blepharitis, anorexia and weakness. Some of them suffered from unilateral blindness and ocular loss. After morphometric analysis, the specimens showed characteristics compatible with the digenean trematode P. gralli. The clinical signs of infection were resolved by manual removal of the adults, treatment with praziquantel and relocation into an environment without a natural water source. In order to determine if an ongoing cycle of this pathogen was present in the shelter, the habitat of the birds was inspected for the presence of infected intermediate hosts and contaminated water and objects. It was found that the snails Melanoides tuberculata acted as the intermediate host, and reared the infectious stages toward other animals, as shown by the reproduction of ocular philophthalmiasis in chickens artificially infected with excysted metacercaria. Moreover, three out of every ten snails found in the place were infected with rediae of P. gralli, raising the possibility of the dispersion of the parasite into new environments as well as the imminent zoonotic risk. The finding of P. gralli in Costa Rica is the first official report in Central America.     

Age differences in kleptoparasitism by laughing gulls (Larus atricilla) on adult and juvenile brown pelicans (Pelecanus occidentalis)

We recorded the choice of victims by a population of adult and first-winter laughing gulls (Larus atricilla) making attempts to steal food from adult and juvenile brown pelicans (Pelecanus occidentalis). Adult gulls made food theft attempts on juvenile pelicans with a disproportionately great frequency; first-winter gulls appeared to select pelican victims at random. Adult pelicans were more frequently successful in their foraging attempts than juvenile pelicans, suggesting that adults may be more reliable as potential food sources for gulls making theft attempts. However, juvenile pelicans attempted to evade kleptoparasitizing gulls less frequently than did adult pelicans, suggesting that the fish prey of juveniles may be more easily stolen. These patterns are discussed in relation to optimal victim choice by gulls, and to deferred maturity in both species.     

Detection, identification and molecular typing of Leishmania major in Phlebotomus papatasi from a focus of zoonotic cutaneous leishmaniasis in central of Iran

Leishmania major is the causative agent and Phlebotomus papatasi is the main vector of rural zoonotic cutaneous leishmaniasis (ZCL) in Iran and elsewhere. Nested PCR protocols were used to amplify a region of the ribosomal RNA amplicon of Leishmania (ITS1-5.8S rRNA gene) in female P. papatasi. In the current investigation, L. major was found in Natanz, Isfahan province in centre of Iran, in a focus of rural ZCL. Ten (1.8%) out of 549 female P. papatasi was found to be infected with L. major based on the PCR detection and sequencing of parasite ITS-rDNA.Nine (1.8%) out of 498 female P. papatasi infected with L. major came from animal shelters, inside houses and yards. And one (1.9%) out of 51 female P. papatasi infected with L. major came from gerbil borrows. Infection rates were higher for females containing red blood meals, large eggs (semi-mature and mature) than for those without either blood meals or eggs. From the 10 infections detected three different haplotypes of L. major were identified. Two haplotypes were found to be novel. The other haplotypes of L. major was found to be identical to that of isolates of L. major from Iran and in elsewhere using GenBank data. Comparisons of infection rates between habitats will be inaccurate when the proportions of blood-fed and gravid flies differ among sandfly samples.     

Colonial breeding reduces nest predation in the common gull (Larus canus)

It is often suggested that colonial breeding reduces nest predation for birds with a high defence capacity, but experimental comparison of predation at solitary and colonial nests is seldom feasible within a single species. We here report on such a test in the common gull (Larus canus). The rate of predation on experimental eggs was significantly lower near colonies than near solitary gull nests, and the eggs survived longer at the edge of a colony than farther away. Communal mobbing of nest predators is the likely reason. In both of two years, almost all nests of solitary gulls were destroyed by predators, while most clutches survived in colonies. Nest predation hence selects strongly for colonial breeding in the present population of common gulls.     

Lymnaea fuscus (Pfeiffer, 1821) as a potential intermediate host of Fascioloides magna in Europe

Experimental infections of two different populations of Lymnaea fuscus in France and Sweden, with a Czech isolate of Fascioloides magna were carried out to determine if this lymnaeid species enables parasite larval development. Species identification of both snail populations was performed using the morphology of the copulatory organ, and also confirmed by sequencing of the internal transcribed spacer 2 (ITS2) region of the snail genomic rDNA. Only juvenile snails measuring less than 3 mm (1–3 weeks of age) were successfully infected (the viable cercariae were recorded) and infection prevalence decreased with age, as documented by increased shell height. In both French and Swedish L. fuscus populations, prevalence ranged between 1.1% and 58.8%. The mean number of metacercariae obtained from cercariae-shedding snails was 13.7 (±11.4), while the total cercarial production noted in snails dissected at day 85 post-exposure was 147.5 (±56.6). Compared to uninfected control snails, we observed reduced growth of infected snails. Despite age-related resistance of snail to the parasite, and limited cercarial production in these experimentally infected snails, F. magna was still able to complete larval development in L. fuscus.     

Effects of body size on sex‐related migration vary between two closely related gull species with similar size dimorphism

Studies of migration have revealed multiple trade‐offs with other life‐history traits that may underlie observed variation in migratory properties among ages and sexes. To assess whether, and to what extent, body size and/or sex‐specific differences in competition for resources (e.g. breeding territories or winter food) may shape variation in migration distance and timing of arrival in ecologically and phylogenetically related species, we combined over 30 000 sightings of individually marked, sexually mature males and females of Herring Gulls Larus argentatus and Lesser Black‐Backed Gulls Larus fuscus with biometric measurements and phenological observations at a mixed breeding colony. In L. argentatus, larger males migrated further from the breeding colony, whereas migration distance was independent of body size in adult females. In L. fuscus, no relationship between body size and migration distance was apparent in either sex. Mean arrival dates at the breeding colony did not vary with migration distances but differed between males and females of L. argentatus (but not L. fuscus). As allometry at least partly explains sexual segregation in migration behaviour in L. argentatus, but not in L. fuscus, we conclude that the effect of body size on sex‐related migratory strategies may vary between closely related, sympatric species despite similar size dimorphism.