Description of Discocotyle ciray n. sp. (Monogenea: Discocotylidae) from Parahucho perryi (Brevoort) from Hokkaido, Japan, with a redescription of D. sagittata (Leuckart, 1842).

Discocotyle sagittata (Leuckart, 1842) (Monogenea: Discocotylidae) is redescribed, based on specimens collected from the type host, Salmo trutta Linnaeus, from the type locality, Freiburg, Germany, supplemented with specimens from S. trutta and rainbow trout, Oncorhynchus mykiss (Walbaum) reared in an Austrian aquarium. The diagnosis of the genus Discocotyle Diesing, 1850 is emended. Discocotyle ciray n. sp. is described, based on immature, preadult and adult specimens from the salmonid, Parahucho perryi (Brevoort) at Eniwa, Hokkaido, Japan. Adult specimens of the new species were about twice as large as those of D. sagittata from S. trutta. When the type specimens of D. ciray n. sp. were examined together with museum specimens from P. perryi at Tsurui, Hokkaido, the body and clamp sizes were positively correlated to the host size. Their measurements from a smaller P. perryi at Tsurui overlapped with those of D. sagittata, showing that these size differences were not suitable differentiating keys. Discocotyle ciray n. sp. can be separated from D. sagittata by the morphologies of the female genital system (relatively anteriorly positioned ovary, short joint vaginal duct and much more strongly winding uterus). The genetic distances of COI mtDNA sequence between D. ciray n. sp. and D. sagittata were 18.0-18.6%. These remarkable genetic divergences also supported the distinct taxonomic status of D. ciray n. sp.

First description of freshwater mite Unionicola sauerensis sp. nov. infesting thick-shelled river mussel Unio crassus.

A sample of 30 thick-shelled river mussels Unio crassus Philipsson (Unionida: Unionidae) was collected from the River Sauer in Luxembourg to acquire data on parasitic infestations of the mussels. Among other parasites, different development stages of freshwater mites were collected from the gills and the mantle of the mussels and were documented with bright-field, stereo, and confocal laser scanning microscopy and microscopic X-ray computed tomography. The retrieved data allowed a morphological description of larvae and female adults of the mites and assigning them to the genus Unionicola Haldeman (Trombidiformes: Unionicolidae) and the subgenus Pentatax Thor. Additionally, adult stages and larvae were barcoded by sequencing a section of the mitochondrial COI and 18S rRNA genes. This resulted in 4 new, similar Unionicola lineages from the adult stages, which differ in at least 14.7% (uncorrected p distance) from those already published. Barcoding of larval DNA was not successful. The comparison with known European species of the genus Unionicola and analysis of the barcoding results allowed the proposal of a new species of the genus Unionicola. The species was named Unionicola sauerensis sp. nov. after the River Sauer in Luxembourg, where the infested mussels were collected.

Koi herpesvirus disease outbreak: Input for the implementation of a surveillance program in South Tyrol - Italy.

A severe episode of high and abnormal mortality was observed in the population of Cyprinus carpio of Lake Caldaro (South Tyrol, Italy) in summer 2016. The diagnostic investigation carried out led to the identification of Koi Herpesvirus (KHV) as the etiologic agent. Following this disease outbreak and its socio-economic consequences, the local authorities, in agreement with the local Fishing Association, decided to implement a surveillance program for the achievement of KHV-free health status (Category I) in the Province, in accordance to the Implementing Decision (EU) 2015/1554. The selected area was a defined geographical compartment (the Monticolo lakes compartment, South Tyrol, Italy), which is located near Lake Caldaro, where the Koi Herpesvirus disease (KHVD) outbreak had occurred. This area is of particular interest because it supplies other water bodies with juvenile C. carpio individuals; with the achievement of a KHV-free health status, South Tyrol could possibly become independent in the breeding of this fish species. Suitable samples were collected and processed during a two-year period in order to detect the presence/absence of KHV. The same samples were tested for other viruses that can affect carp, namely spring viraemia of carp (SVCV) and carp edema virus (CEV). According to the results, the authors conclude that the Monticolo lakes area should be classified as KHV-free, as no sample has tested positively for the presence of this specific virus (KHV).

Acanthocephalan parasites collected from Austrian fishes: molecular barcoding and pathological observations.

Acanthocephalan parasites were collected from the intestinal tracts of 137 predominantly wild fish (1 barbel Barbus barbus, 3 European chub Squalius cephalus, 13 rainbow trout Oncorhynchus mykiss and 120 brown trout Salmo trutta) from 12 localities. The condition factor, intensity of acanthocephalan infection and pathological lesions, if applicable, were documented. Routine bacteriology and virology were performed, and the brown trout were additionally tested for the presence of the myxozoan parasite Tetracapsolioides bryosalmonae by PCR. In total, 113 acanthocephalans were barcoded by sequencing a section of the mitochondrial cytochrome oxidase subunit I (COI) gene. Barcoding of the acanthocephalan tissues resulted in 77 sequences, of which 56 were assigned to Echinorhynchus truttae (3 genotypes), 11 to Pomphorhynchus tereticollis (9 genotypes), 9 to Acanthocephalus sp. (5 genotypes) and 1 to Neoechinorhynchida. Most of these genotypes were detected for the first time. Statistically, the acanthocephalan infection did not have an impact on the condition factor of the brown trout. Infection with P. tereticollis caused more severe pathological changes in the digestive tract than E. truttae. The present study provides new data regarding the distribution of acanthocephalan species in Austria and their impact on individual fish. In addition, new barcoding data from acanthocephalan parasites are presented, and the occurrence of P. tereticollis in European chub in Austria and in brown and rainbow trout in general was confirmed for the first time.

Identification of new genogroups in Austrian carp edema virus isolates.

Carp edema virus disease (CEVD), also known as koi sleepy disease (KSD), is an emerging viral disease caused by the carp edema virus (CEV). It is characterized by lethargic behavior, gill necrosis, and generalized edema, leading to significant morbidity and mortality in common carp and koi Cyprinus carpio. Accurate diagnosis of CEVD relies on amplification of a P4a protein-encoding DNA segment from the CEV genome. A phylogenetic analysis of amplified fragments revealed 3 distinct CEV genogroups: I, IIa, and IIb. We explored the phylogenetic relationship between Austrian CEV isolates with existing CEV genogroups. The phylogenetic analysis (n = 18) established the presence of the 3 extant CEV genogroups as well as 2 new CEV genogroups (IIIa and IIIb) classified to identify the Austrian isolates that were distinct from the existing CEV genogroups. It is evident that CEV infection cases are growing in number each year, which may be due to development of sensitive diagnostic assays, while information regarding the virus is scarce. National and international efforts are required to study the epidemiology of the CEV in major carp-producing countries.

Hole-in-the-head disease in discus fish, Symphysodon (Heckel, 1840): Is it a consequence of a dietary Ca/P imbalance?

Hole-in-the-head (HITH) disease-affected fish develop characteristic lesions in the skin above sensory pores of the head and the trunk. This study investigated whether an unfavourable Ca/P ratio in the diet could provoke lesions consistent with HITH disease in discus fish Symphysodon (Heckel, 1840) as a comparable condition to secondary hyperparathyroidism of tetrapod species. Two groups of five fish were fed a plain beef heart diet (Ca/P of 0.03), whereas two other groups were kept on commercial discus feed (Ca/P of 2.73). Each feeding group was submitted to two different water hardness regimes (35.66-71.39 mg/L CaCO3 and 124.94-196.33 mg/L CaCO3 , respectively). All fish were observed for the development of the characteristic lesions for 16 weeks. At the end of the study, histological, bacteriological and parasitological examinations were conducted and plasma Ca, P and Mg values were determined. Diplomonad flagellates were detected in two fish. Isolated bacteria of all groups mostly belonged to Aeromonadales and Pseudomonadales. No significant difference of plasma mineral values between the groups was observed. Compared to the results of other authors, Ca stayed mainly in the range and P exceeded the reference values. Histological examinations did not indicate HITH disease, and no fish developed signs of the disease during the study. Clinical trial registration number GZ 68.205/0135-WF/V/36/2014.

PRELIMINARY RADIOGRAPHIC OBSERVATIONS OF VERTEBRAL CHANGES IN DISCUS ( SYMPHYSODON DISCUS).

Discus ( Symphysodon discus) maintained in aquaria are held in a wide range of water parameters and subjected to many different feeding regimes. In a pilot study, four groups of discus (mean length 12.1 cm, mean weight 57.3 g) were submitted to a radiographic examination to assess the skeletal structure of the vertebral column under defined environmental conditions. Water temperature was 30°C for all groups. Two groups were held at <28.6 mg/L calcium (Ca) and two at 50.0-78.6 mg/L Ca within the ambient water. One of each water quality group was fed a commercial discus diet while the other two groups were kept on a plain beef-heart diet, creating a total of four separate groups. In the case of the beef heart group, dietary Ca content (g/kg) was 0.06 and phosphorous (P) content 2.06, leading to a Ca : P ratio of 0.03, whereas in the commercial diet group a Ca content of 20.1 g/kg and P of 7.36 g/kg resulted in a Ca : P ratio of 2.7. Magnesium (Mg) contents of the beef-heart diet were 0.21 and of the commercial diet 1.69 g/kg. Six fish were submitted to radiography at the beginning of the experiment as a control. After 16 wk of the above diets and environmental conditions, radiographs were taken from all fish (six per group) and evaluated by three independent persons using a scoring system. Alterations were found in all groups. The results of this pilot study give reason to scrutinize rearing and keeping conditions of this fish species.

Distribution and prevalence of T. bryosalmonae in Austria: A first survey of trout from rivers with a shrinking population.

The first evidence of proliferative kidney disease (PKD) in an Austrian river (the River Kamp) was documented in 2016, and no information on the PKD infection status of trout in other rivers was available. Since then, brown trout (Salmo trutta fario) and rainbow trout (Oncorhynchus mykiss) have been collected from rivers in Upper and Lower Austria for different diagnostic purposes. In this study, we summarize the recent findings of a first survey concerning the distribution of Tetracapsuloides bryosalmonae, the causative agent of proliferative kidney disease (PKD), from these samples. Between September 2015 and October 2017, a total of 280 brown trout and 39 rainbow trout were collected from 21 rivers in the provinces of Upper and Lower Austria. T. bryosalmonae was detected by PCR of kidney tissue in 17 of 21 sampled rivers and in 138 of 280 brown trout as well as in 11 of 39 rainbow trout. Pathological signs of PKD (e.g., hypertrophy of the kidney) were observed in 33 analysed brown trout and six rainbow trout samples. No correlations between fish infected by T. bryosalmonae and the parameters size and age class, condition factor, geological origin of the streams and distribution within the river course were found, while positively tested fish are significantly increased at sampling sites exceeding water temperatures of 15°C for median periods of 115 days. The prevalence within the affected streams or stream sections is highly variable, and in single rivers, infection rates of up to 90% are confirmed.

Morphological and molecular characterization of Thelohanellus hoffmanni sp. nov. (Myxozoa) infecting goldfish Carassius auratus auratus.

A new species of the genus Thelohanellus Kudo, 1933 (Myxosporea, Bivalvulida) was isolated from the fins of goldfish Carassius auratus auratus (Linnaeus 1758). The fish had been imported from China by an Austrian retailer. Nodules from the margins of the fins contained pyriform myxospores with a singular polar capsule. In valvular view, the spores measured 12.2 µm in length and 6.4 µm in width. In sutural view, the thickness was 2.9 µm. The polar capsule measured 4.2 × 3.1 µm and contained a polar filament with 8 to 9 coils. Histological sections showed plasmodia of 0.2 to 4.0 mm diameter with the earlier developmental stages of the parasite in the periphery and the mature spores closer to the center. In the transmission electron microscope examination, the different developmental stages could be observed. Morphological data, host specificity, tissue tropism, and molecular analysis of the small subunit rDNA identify this parasite as a new species of Thelohanellus, which we have named Thelohanellus hoffmanni sp. nov.