Non-invasive detection of Orthohalarachne attenuata (Banks, 1910) and Orthohalarachne diminuata (Doetschman, 1944) (Acari: Halarachnidae) in free-ranging synanthropic South American sea lions Otaria flavescens (Shaw, 1800).

Respiratory mites of the genera Orthohalarachne and Halarachne (Acari: Halarachnidae) are causative agents of nasopharyngeal/nasopulmonary acariasis in pinnipeds and sea otters. Until now, these endoparasitic mites were mainly diagnosed via necropsies and invasive procedures. So far, non-invasive diagnostic techniques have neither been developed nor applied in free-ranging pinnipeds. In the current study, we aimed to evaluate the halarachnid mite infestation status of free-ranging "urban" South American sea lions Otaria flavescens in the city of Valdivia, Chile. Therefore, non-invasive sampling methods were used in the current study, e. g. by observation-based sampling of freshly expectorated nasal mucus in the animal environment. Further, collection devices were developed for target-oriented sampling of sneezed nasal mucus, including sterile petri dishes and stretched clingfilms mounted on telescopic rods. Applying these techniques, 26 individual sputum samples were collected. 11.5% of sputum samples proved positive for halarachnid larvae (in total, n = 7), which were morphologically identified as Orthohalarachne attenuata (n = 2) or Orthohalarachne diminuata (n = 5). In one of the individual sea lion mucus samples, both Orthohalarachne species were detected, thereby confirming a patent co-infestation in vivo. 16S rDNA-based molecular identification of individual Orthohalarachne spp. larvae confirmed morphological findings. For the first time, we here molecularly characterized Orthohalarachne spp. on the basis of three gene regions [18S, 28S and the internal transcribed spacer 1 (ITS1)]. Overall, current data include the successful application of non-invasive techniques to sample halarachnid mites from free-ranging synanthropic pinnipeds and contribute to the current knowledge on respiratory mites infesting South American sea lions by combining morphological and molecular methods to overcome challenges in species identification. This study should further serve as baseline study and calls for more research on occurrence, biology and health implications of orthohalarachnosis in free-living as well as captive pinnipeds.

First report of a severe nasopulmonary acariasis caused by Orthohalarachne diminuata Doetschman, 1944 (Acari: Halarachnidae) in a captive South American sea lion (Otaria flavescens Shaw, 1800).

Obligatory endoparasitic mites of the genera Halarachne Allman, 1847 and Orthohalarachne Newell, 1947 (Acari: Halarachnidae) parasitize different segments of the respiratory tract of marine mammals, including pinnipeds and sea otters, and infestations can cause asymptomatic to serious respiratory diseases. However, knowledge on biology, pathogenic potential and occurrence of halarachnid mites infesting pinnipeds, especially in captivity, is scarce. A two-year-old South American sea lion (Otaria flavescens Shaw, 1800) male, born and held at the Vienna Zoo, was anesthesized for routine pre-transport examinations, including computed tomography, bronchoalveolar lavage, and blood sampling. During the final phase of general anesthesia, the individual abruptly became apneic and died despite all attempts at resuscitation. At necropsy, 45 highly motile whitish millimeter-sized structures were macroscopically detected in the trachea, bifurcatio tracheae and main bronchi and were identified as adult stages of Orthohalarachne diminuata Doetschman, 1944 following morphological descriptions. After trepanation of the nasal cavity and sinus paranasalis, a total of 407 larval and 3 nymphal specimens distributed in clusters were detected. Macroscopically, sinus mucosa showed hyperemia and multiple petechial hemorrhages. Histopathological analyses of paranasal sinuses revealed mite cross-sections surrounded by sanioserous exudate and epithelial exfoliation. For the first time, O. diminuata was molecularly characterized and phylogenetically analyzed based on its 16S rDNA. Our study constitutes the first record of a severe O. diminuata infestation in captive O. flavescens and one of the few host-parasite records in general. We present clinical data and pathological results, the first scanning electron microscopic images of a O. diminuata larval stage and discuss the etiology of this autochthonous infestation, possible transmission pathways and detrimental effects. Further studies on biology and pathogenic effects of halarachnid mites, as well as on the development of non-invasive sampling techniques are essentially required for a better understanding of (ortho-)halarachnosis in pinnipeds held in zoological gardens.

Genetic diversity of Fasciola hepatica in Austria.

The worldwide occurring common liver fluke Fasciola hepatica can infect humans and animals and leads to considerable illness and economic loss annually. The aim of this study was to determine the genetic diversity of F. hepatica in Austria. In total, 31 adult flukes isolated from cattle from various regions in Austria were investigated for their cytochrome oxidase subunit 1 (cox1) and nicotinamide dehydrogenase subunit 1 (nad1) gene sequences. It was shown that Austrian isolates of F. hepatica reveal extensive genetic diversity. To the best of our knowledge, these are the first data on the diversity of F. hepatica in Austria.

Wherefrom and whereabouts of an alien: the American liver fluke Fascioloides magna in Austria: an overview.

The giant liver fluke Fascioloides magna, an invasive species originating from North America, was recorded in Austria in the wild for the first time in 2000. Since then, various data concerning the epidemiology in snail intermediate hosts and cervid final hosts have been reported. Galba truncatula acts as snail intermediate host, and red deer, roe deer and fallow deer act as final hosts. G. truncatula is abundant throughout the region, especially along muddy shores of slow-flowing branches of the river system. Prevalence in deer (20-100 %) is much higher than in snails (0.03-0.2 %). Despite medical treatment of parts of the deer population, the parasite has successfully established itself on both sides of the Danube floodplain environments southeast of Vienna. Genetic analysis revealed that the infection of Austrian deer populations apparently originated from foci in the Czech Republic or from populations of Danube tributaries. Areas adjacent southwards, which will soon be joined through wildlife crossings, have not yet evidenced F. magna. Nonetheless, these environments are inhabited by host snails and deer and therefore constitute suitable habitats for F. magna. Invading alien parasites not only threaten native individual hosts but also influence host populations, thus potentially also modifying parasite communities and interactions. The host range of F. magna includes a variety of potential hosts, notably other Lymnaeidae as potential intermediate hosts and various ungulates, including sheep and cattle, as final hosts. Because eradication after medical treatment was unsuccessful, and due to the risk of further spread of the parasite into unaffected regions, enhanced control strategies need to be developed. We recommend assessment of introduction pathways and dispersal, continuous monitoring of host abundance and distribution and the prevalence of flukes in intermediate and final hosts, as well as coordinated and concerted actions with neighbouring countries. This strategy could help to reduce potential negative impacts of this and other invasive parasites on host populations in Europe.

Fasciola hepatica - monitoring the milky way? The use of tank milk for liver fluke monitoring in dairy herds as base for treatment strategies.

In this study 595 lactating cows originating from 31 carinthian farms were investigated in accordance of liver fluke infection using individual and tank milk as well as individual blood and faecal samples. Two commercial ELISAs were used to test the milk and blood serum, and the results were compared with coproscopy and a commercial copro-antigen ELISA. Receiver operating characteristics (ROC) and two-graph operating characteristics (TG ROC) of tank milk results were conducted based on the individual milk to determine the minimum reliable in-herd antibody prevalence for the predominant condition in the investigation area. In 17.8% of the examined individuals located in 64.5% of the farms eggs were detected by coproscopy. The copro-antigen ELISA delivered 13.4% positive individuals from 54.8% of the farms. The milk ELISAs showed 42.7% (Euroclone) and 44.2% (Pourquier) positive cows on 90.3% of the farms. The blood samples were positive in 43% (Euroclone) and 45.2% (Pourquier) of the individuals from 90.3% to 96.8% of the herds, respectively. Based on the milk and the blood an average in-herd prevalence of 30-45% can be assumed. The serum and milk samples delivered correlating results with kappa values between 0.94 and 0.97, whereas the coproscopy and copro-antigen ELISA did not correlate well with the ELISA results. The two different ELISA tests highly correlated on individual and on herd level. Both showed a reliable minimum in-herd prevalence of ∼20%, meaning that one fifth of the individuals in a herd have to be positive to obtain a positive bulk tank milk result. In the investigated area a higher in-herd prevalence is expected, therefore the tank milk is useful as a monitoring tool and can be used as a basis for intervention strategies.

[Incidence and control of the American giant liver fluke, Fascioloides magna, in a population of wild ungulates in the Danubian wetlands east of Vienna]. / Vorkommen und Bekämpfung des Amerikanischen Riesenleberegels, Fascioloides magna, in einer Schalenwildpopulation in den Donauauen östlich von Wien.

Fascioloides magna,the giant liver fluke, is an introduced parasite imported to Europe with infected game which is endemic in the Austrian wetlands of the Danube from Vienna to the Slovakian border. Due to its pathogenicity (especially in roe deer) and its potential of transmission to domestic ruminants a project on the epidemiology and control of this parasite was carried out between 2000 and 2005. To assess distribution faecal droppings from red deer were collected in an area along the Danube and livers of red and roe deer shot or found dead were examined. Simultaneously, triclabendazole-medicated feed was offered repeatedly in several areas of known fluke presence from 2001. The average prevalence of infection as demonstrated by Fascioloides eggs in droppings (n = 145) was 6.3%, with > 50% in some areas. Infection rates decreased from 11.3% (01/2001) to 1.5% (04/2002) in the whole area. In Fischamend, a highly endemic area, prevalences in monthly samples (n = 25) decreased from 70% (01/2001) to less than 1% (01/2005). Similarly, the number of eggs per gram of faeces was significantly reduced. Of 457 red deer livers 15.8% were positive. In the Fischamend area, a reduction of positive livers was also found (2000:100%; 2005:13%), as well as a reduction of the numbers of flukes/liver. Control of F. magna in the Danubian wetlands by anthelmintic treatment proved to be successful although eradication was not achieved.

Effects of excretory/secretory products of Haemonchus contortus on cell vacuolation.

Excretory/secretory (ES) products of the gastric nematode, Haemonchus contortus, have been implicated in the inhibition of gastric acid secretion which follows infection. Parietal cell vacuolation has been observed in abomasal sections from parasitised sheep, and ES prepared in vitro has been reported to cause vacuolation and to increase neutral red (NR) uptake in epithelial cell cultures. We have used the latter approach to examine, at the cellular level, the effects of ES prepared from L3 and adult nematodes. Both NR uptake and cellular vacuolation were increased following exposure to larval or adult ES products. ES preparations from adult worms induced more extensive vacuolation then those from L3 worms and, as with VacA treatment, adherent cells remained viable despite high levels of vacuolation. Whereas VacA-induced vacuolation resulted in NR uptake predominantly localised in vacuoles, this appeared not to be the case with ES-induced vacuolation, suggesting that different mechanisms might be involved. Both ES and VacA exposure was associated with an increased rate of cell detachment.

[Holiday dreams with a downside: uncommon tapeworm infection in an infant]. / Traumurlaub mit Folgen -- ein ungewöhnlicher Bandwurmbefall bei einem zweieinhalbjährigen Kind.

Eight weeks after his return from Mauritius, a 2.5-year-old boy started to show signs of gastrointestinal illness including diarrhoea with frequent ill-smelling stools as well as restlessness, insomnia and apathy. Proglottids excreted with the faeces were diagnosed as Raillietina madagascariensis (Davain, 1870), the tropical rat tapeworm. Treatment with praziquantel (10 mg/kg BW) proved to be efficient.

Scraping or shaking--a comparison of methods for the quantitative determination of Echinococcus multilocularis in fox intestines.

A modified sedimentation technique (the "shaking in a vessel" technique; SVT) to examine intestines for smaller helminths such as Echinococcus multilocularis is described and compared with the intestinal scraping technique (IST). Out of 356 foxes, 26 were E. multilocularis positive, 19 using IST (sensitivity 73.1%) and 25 with SVT (sensitivity 96.2%) after IST. Especially low infection intensities went undetected with IST. The result is comparable to the established sedimentation and counting technique (SCT). SVT is recommended for the routine examination of fox intestines as an alternative to SCT.

[Screening for antibodies against zoonotic agents among employees of the Zoological Garden of Vienna, Schönbrunn, Austria]. / Untersuchungen auf Antikörper gegen Zoonoseerreger bei Angestellten des Wiener Tiergartens Schönbrunn.

The aim of this study was to investigate the prevalence of antibodies against zoonotic agents in employees of the zoological garden of Vienna, Schönbrunn, Austria. Sixty out of 120 employees participated in the study. In 97% of them antibodies to at least one zoonotic agent were identified. Only two participants were free of antibodies to the zoonotic agents tested. The following seroprevalences (in brackets) were obtained: Viral zoonotic (and potentially zoonotic) agents: Influenzavirus A/H1N1 (58%), Influenzavirus A/H3N2 (85%), Lymphocytic choriomeningitis virus (13%), Encephalomyocarditis virus (5%), Orthopox- (Cowpox-) virus and Hantavirus type Puumala (3%). Hantavirus type Hantaan and Borna disease virus (all negative). Bacterial zoonotic agents: Bartonella henselae (65 %), Borrelia burgdorferi (10%), Leptospira interrogans serovar copenhageni and serovar icterohaemorrhagiae as well as Chlamydophila psittoci (2% each). Brucella spp., Coxiella bumetii, and Francisella tularensis (all negative). Parasitic zoonotic agents: Toxoplasma gondii (53%), Toxocara spp. (21%), Capillaria hepatica (2%), Fasciola hepatica, Schistosoma mansoni, E. multilocularis, and E. granulosus (all negative). The remarkably high seroprevalence to the causative agent of cat scratch disease, Bartonella henselae, is probably due to the private contact of the employees to cats. Regarding viral zoonotic agents it has to be mentioned that Influenzavirus vaccination and/or human-to-human transmission of especially A/H3N2 Influenzaviruses probably attributed significantly to the very high seroprevalence to both Influenzavirus types A/H1N1 and A/H3N2. When investigating parasitic zoonotic agents, high prevalence rates were found against Toxoplasma gondii and Toxocara spp., however, it was not possible to establish a causal link between seropositivity and the professional activity in the zoo. Interestingly, in the case of antibodies to T. gondii, the typical correlation with age was not found in this study, while in the case of the Toxocara spp. positive subjects a correlation was identified with both age and duration of employment in the zoo. Regarding the later two zoonotic parasites, employees of the zoological garden showed significantly higher seroprevalences than the average Austrian population. Antibodies to Capillaria hepatica, a hepatic-parasite in rodents which is diagnosed in humans rarely, were identified in one employee and another one showed a questionable positive result. Further investigations did not exhibit clinical infestations with the parasite in these two individuals so far.

[Ophthalmomyiasis caused by Oestrus ovis in a 3 year old boy]. / Oestrus ovis als Ursache einer Ophthalmomyiasis bei einem 3-jährigen Kind.

Oestrus ovis larvae are obligatory parasites of the nasal and sinus cavities of sheep and goats. Oestrosis in small ruminants is prevalent in Mediterranean and tropical countries. Occasionally, humans are affected by ocular myiasis (ophthalmomyiasis) and more rarely, nasal myiasis. A case of human ophthalmomyiasis caused by first instar larvae of Oestrus ovis in a 3 year old boy was observed in Gran Canaria. Acute catarrhal conjunctivitis developed within few hours. During the inspection of the eye several larvae could be observed, moving quickly over the cornea to the conjunctival sac. Rinsing and mechanical removal of the larvae lead to an uncomplicated recovery.