Allopodocotyle palmi sp. nov. and Prosorhynchus maternus Bray & Justine, 2006 (Digenea: Opecoelidae & Bucephalidae) from the Orange-Spotted Grouper Epinephelus coioides (Hamilton, 1822) off Bali, Indonesia, Described Using Modern Techniques.

BACKGROUND: The most convincing species of Allopodocotyle Pritchard, 1966 (Digenea: Opecoelidae) are known overwhelmingly from groupers (Serranidae: Epinephelinae). Six species of Allopodocotyle have been reported, collectively, from species of Cromileptes Swainson, 1839, Epinephelus Bloch, 1793 and Plectropomus Oken, 1817. These are A. epinepheli (Yamaguti, 1942), A. heronensis Downie & Cribb, 2011, A. manteri (Saoud & Ramadan, 1984), A. mecopera (Manter, 1940), A. plectropomi (Manter, 1963) and A. serrani (Yamaguti, 1952). In addition, a not yet fully described and unnamed seventh species, morphologically and phylogenetically close to A. epinepheli, was isolated from the orange-spotted grouper Epinephelus coioides (Hamilton, 1822) off Bali, Indonesia in 2016. An eighth species, again from E. coioides off Bali is described herein. METHODS: Morphological and phylogenetic analyses justify the recognition of A. palmi sp. nov., which is also genetically different from the as yet unnamed congener from the same host and locality. For the first time, 3D confocal laser scanning microscopy was applied to study and distinguish Digenea taxonomically. We introduce the 'Palm pattern', a new simplified way to visualise morphometric differences of related digenean taxa. RESULTS: Allopodocotyle palmi sp. nov. is distinguished from its congeners that infect groupers by its elongate body with a size > 2.7 mm and diagonal testes. The ovary is located mainly, and the anterior testis completely, in the posterior half of the body; the uterine coils are in the fourth eighth of the body. The cirrus-sac is 0.75-1.4 (1.1) mm long, its posterior extremity is well separated from the anterior extent of the vitelline fields, just reaching the anterior border of uterine coils. In addition, Prosorhynchus maternus Bray & Justine, 2006 (Bucephalidae) was isolated from E. coioides, representing the first record in Indonesia and the third record for this fish species. CONCLUSION: The biodiversity research in Indonesia is enhanced with a new species description based on modern and newly applied techniques.

Metazoan endoparasite fauna and feeding ecology of commercial fishes from Java, Indonesia.

Despite being an important component of the marine ecosystem and posing health risks to human seafood consumers, fish parasites in Indonesia have yet to be adequately described. Here, we analyzed the diet and metazoan parasite fauna of seven commercial fish species (Alectis indica, Carangoides chrysophrys, Johnius borneensis, Mene maculata, Trichiurus lepturus, Upeneus asymmetricus, U. moluccensis) landed in Java, Indonesia. We isolated 11 endoparasite species, established 22 new host and 14 new locality records, and extended parasitological records of A. indica by 24%, C. chrysophrys by 25%, J. borneensis by 40%, M. maculata by 44%, U. asymmetricus by 100%, and U. moluccensis by 17%. We genetically identified the trematode Stephanostomum cf. uku (of Bray et al. 2005) from Alecta indica for the first time in Indonesia and provided the sequence of its 28S marker. Stomach content analysis revealed seven different prey items, and the examined fish species were grouped into four feeding categories, which differed significantly in their respective endoparasite fauna. All but two examined fish species hosted potentially zoonotic nematodes, which reveal a risk for parasite-borne diseases in Indonesian food fishes and call for more consequent monitoring with regard to seafood safety in this region. With this study, we were able to establish an association between the feeding ecology and the endoparasite fauna of marine fishes which will help to better understand the transmission pathways of (potentially zoonotic) parasites in food fishes in tropical waters.

Practical measures for sustainable shark fisheries: Lessons learned from an Indonesian targeted shark fishery.

Overfishing is a major threat to the survival of shark species, primarily driven by international trade in high-value fins, as well as meat, liver oil, skin and cartilage. The Convention on the International Trade in Endangered Species of Wild Fauna and Flora (CITES) aims to ensure that commercial trade does not threaten wild species, and several shark species have recently been listed on CITES as part of international efforts to ensure that trade does not threaten their survival. However, as international trade regulations alone will be insufficient to reduce overexploitation of sharks, they must be accompanied by practical fisheries management measures to reduce fishing mortality. To examine which management measures might be practical in the context of a targeted shark fishery, we collected data from 52 vessels across 595 fishing trips from January 2014 to December 2015 at Tanjung Luar fishing port in East Lombok, Indonesia. We recorded 11,920 landed individuals across 42 species, a high proportion of which were threatened and regulated species. Catch per unit effort depended primarily on the number of hooks and type of fishing gear used, and to a lesser degree on month, boat engine power, number of sets and fishing ground. The most significant factors influencing the likelihood of catching threatened and regulated species were month, fishing ground, engine power and hook number. We observed significant negative relationships between standardised catch per unit effort and several indicators of fishing effort, suggesting diminishing returns above relatively low levels of fishing effort. Our results suggest that management measures focusing on fishing effort controls, gear restrictions and modifications and spatiotemporal closures could have significant benefits for the conservation of shark species, and may help to improve the overall sustainability of the Tanjung Luar shark fishery. These management measures may also be applicable to shark fisheries in other parts of Indonesia and beyond, as sharks increasingly become the focus of global conservation efforts.

Pseudempleurosoma haywardi sp. nov. (Monogenea: Ancyrocephalidae (sensu lato) Bychowsky & Nagibina, 1968): An endoparasite of croakers (Teleostei: Sciaenidae) from Indonesia.

An endoparasitic monogenean was identified for the first time from Indonesia. The oesophagus and anterior stomach of the croakers Nibea soldado (Lacépède) and Otolithes ruber (Bloch & Schneider) (n = 35 each) sampled from the South Java coast in May 2011 and Johnius amblycephalus (Bleeker) (n = 2) (all Sciaenidae) from Kedonganan fish market, South Bali coast, in November 2016, were infected with Pseudempleurosoma haywardi sp. nov. Prevalences in the first two croakers were 63% and 46%, respectively, and the two J. amblycephalus harboured three and five individuals. All three croakers represent new hosts for this monogenean genus. We provide infection rates, light microscopical observations, 3D confocal microscopical illustrations, and a morphometric comparison with all congeners. The new species differs in body size, the position and shape of the ovary and testes, and especially in the composition of the dorsal anchor complex, with the dorsal bar being anteriorly concave rather than planar or convex as in its congeners. The dorsal and ventral anchors of this new species are the longest in the genus, whereas the male copulatory organ is the smallest. The first DNA sequences for a member of this genus demonstrate the greatest similarity with endoparasitic freshwater monogeneans from African cichlid fishes. This suggests a freshwater origin for these marine endoparasitic monogeneans.

Protozoan and helminth parasite fauna of free-living Croatian wild wolves (Canis lupus) analyzed by scat collection.

The European wolf (Canis lupus) is a large carnivore species present in limited areas of Europe with several small populations still being considered as endangered. Wolves can be infected by a wide range of protozoan and metazoan parasites with some of them affecting free-living wolf health condition. On this account, an epidemiological survey was conducted to analyze the actual parasite fauna in Croatian wild wolves. In total, 400 individual faecal samples were collected during field studies on wolf ecology in the years 2002-2011. Parasite stages were identified by the sodium acetate acetic acid formalin (SAF)-technique, carbolfuchsin-stained faecal smears and Giardia/Cryptosporidium coproantigen-ELISAs. A subset of taeniid eggs-positive wolf samples was additionally analyzed by PCR and subsequent sequencing to identify eggs on Echinococcus granulosus/E. multilocularis species level. In total 18 taxa of parasites were here detected. Sarcocystis spp. (19.1%) occurred most frequently in faecal samples, being followed by Capillaria spp. (16%), ancylostomatids (13.1%), Crenosoma vulpis (4.6%), Angiostrongylus vasorum (3.1%), Toxocara canis (2.8%), Hammondia/Neospora spp. (2.6 %), Cystoisospora ohioensis (2.1%), Giardia spp. (2.1%), Cystoisospora canis (1.8%), Cryptosporidium spp. (1.8%), Trichuris vulpis (1.5%), Taenia spp. (1.5%), Diphyllobothrium latum (1.5%), Strongyloides spp. (0.5%), Opisthorchis felineus (0.5%), Toxascaris leonina (0.3%), Mesocestoides litteratus (0.3%) and Alaria alata (0.3%). Some of the here identified parasites represent relevant pathogens for wolves, circulating between these carnivorous definitive hosts and a variety of mammalian intermediate hosts, e. g. Taenia spp. and Sarcocystis spp., while others are considered exclusively pathogenic for canids (e.g. A. vasorum, C. vulpis, T. vulpis, Cystoisospora spp.). This study provides first records on the occurrence of the two relevant anthropozoonotic parasites, Giardia spp. and Cryptosporidium spp., in wild wolves from Croatia.

Microbial Diversity and Parasitic Load in Tropical Fish of Different Environmental Conditions.

In this study we analysed fecal bacterial communities and parasites of three important Indonesian fish species, Epinephelus fuscoguttatus, Epinephelus sexfasciatus and Atule mate. We then compared the biodiversity of bacterial communities and parasites of these three fish species collected in highly polluted Jakarta Bay with those collected in less polluted Indonesian areas of Cilacap (E. sexfasciatus, A. mate) and Thousand Islands (E. fuscoguttatus). In addition, E. fuscoguttatus from net cages in an open water mariculture facility was compared with free living E. fuscoguttatus from its surroundings. Both core and shared microbiomes were investigated. Our results reveal that, while the core microbiomes of all three fish species were composed of fairly the same classes of bacteria, the proportions of these bacterial classes strongly varied. The microbial composition of phylogenetically distant fish species, i.e. A. mate and E. sexfasciatus from Jakarta Bay and Cilacap were more closely related than the microbial composition of more phylogentically closer species, i.e. E. fuscoguttatus, E. sexfasciatus from Jakarta Bay, Cilacap and Thousand Islands. In addition, we detected a weak negative correlation between the load of selected bacterial pathogens, i.e. Vibrio sp. and Photobacterium sp. and the number of endoparasites. In the case of Flavobacterium sp. the opposite was observed, i.e. a weak positive correlation. Of the three recorded pathogenic bacterial genera, Vibrio sp. was commonly found in E. fuscoguttatus from mariculture, and lessly in the vicinity of the net cages and rarely in the fishes from the heavily polluted waters from Jakarta Bay. Flavobacterium sp. showed higher counts in mariculture fish and Photobacteria sp. was the most prominent in fish inside and close to the net cages.

Parasite fauna of Epinephelus coioides (Hamilton, 1822) (Epinephelidae) as environmental indicator under heavily polluted conditions in Jakarta Bay, Indonesia.

The objective of this study was to assess the environmental conditions of a heavily polluted marine habitat using descriptors of fish parasites. Epinephelus coioides from Jakarta Bay as well as off Jakarta Bay was studied for metazoan parasites. Based on 70 fish and considering previous studies (230 fish), an environmental indicator system was designed. Including the recent study, a total of 51 parasite species have been recorded for E. coioides in Indonesian waters. Seven of them combined with five parasitological indices are useful descriptors for the environmental status of marine ecosystems. The results are visualized in a star graph. A significant different parasite infection between nine analyzed localities demonstrates the negative influence of the megacity Jakarta onto the coastal environment. We herewith complete a parasite based indicator system for Indonesian coastal waters, and suggest that it can be used in other marine habitats as well as for further epinephelids.

Accumulation patterns of lipophilic organic contaminants in surface sediments and in economic important mussel and fish species from Jakarta Bay, Indonesia.

Non-target screening analyses were conducted in order to identify a wide range of organic contaminants in sediment and animal tissue samples from Jakarta Bay. High concentrations of di-iso-propylnaphthalenes (DIPNs), linear alkylbenzenes (LABs) and polycyclic aromatic hydrocarbons (PAHs) were detected in all samples, whereas phenylmethoxynaphthalene (PMN), DDT and DDT metabolites (DDX) were detected at lower concentrations. In order to evaluate the uptake and accumulation by economic important mussel (Perna viridis) and fish species, contaminant patterns of DIPNs, LABs and PAHs in different compartments were compared. Different patterns of these contaminant groups were found in sediment and animal tissue samples, suggesting compound-specific accumulation and metabolism processes. Significantly higher concentrations of these three contaminant groups in mussel tissue as compared to fish tissue from Jakarta Bay were found. Because P. viridis is an important aquaculture species in Asia, this result is relevant for food safety.

Endoparasite survey of free-swimming baleen whales (Balaenoptera musculus, B. physalus, B. borealis) and sperm whales (Physeter macrocephalus) using non/minimally invasive methods.

A number of parasitic diseases have gained importance as neozoan opportunistic infections in the marine environment. Here, we report on the gastrointestinal endoparasite fauna of three baleen whale species and one toothed whale: blue (Balaenoptera musculus), fin (Balaenoptera physalus), and sei whales (Balaenoptera borealis) and sperm whales (Physeter macrocephalus) from the Azores Islands, Portugal. In total, 17 individual whale fecal samples [n = 10 (B. physalus); n = 4 (P. macrocephalus); n = 2 (B. musculus); n = 1 (B. borealis)] were collected from free-swimming animals as part of ongoing studies on behavioral ecology. Furthermore, skin biopsies were collected from sperm whales (n = 5) using minimally invasive biopsy darting and tested for the presence of Toxoplasma gondii, Neospora caninum, and Besnoitia besnoiti DNA via PCR. Overall, more than ten taxa were detected in whale fecal samples. Within protozoan parasites, Entamoeba spp. occurred most frequently (64.7%), followed by Giardia spp. (17.6%) and Balantidium spp. (5.9%). The most prevalent metazoan parasites were Ascaridida indet. spp. (41.2%), followed by trematodes (17.7%), acanthocephalan spp., strongyles (11.8%), Diphyllobotrium spp. (5.9%), and spirurids (5.9%). Helminths were mainly found in sperm whales, while enteric protozoan parasites were exclusively detected in baleen whales, which might be related to dietary differences. No T. gondii, N. caninum, or B. besnoiti DNA was detected in any skin sample. This is the first record on Giardia and Balantidium infections in large baleen whales.

Endo- and ectoparasites of large whales (Cetartiodactyla: Balaenopteridae, Physeteridae): Overcoming difficulties in obtaining appropriate samples by non- and minimally-invasive methods.

Baleen and sperm whales, belonging to the Order Cetartiodactyla, are the largest and heaviest existent mammals in the world, collectively known as large whales. Large whales have been subjected to a variety of conservation means, which could be better monitored and managed if physiological and pathophysiological information, such as pathogen infections, could already be gathered from free-swimming animals instead of carcasses. Parasitic diseases are increasingly recognized for their profound influences on individual, population, and even ecosystem health. Furthermore, a number of parasite species have gained importance as opportunistic neozoan infections in the marine environment. Nonetheless, traditional approaches to study parasitic diseases have been impractical for large whales, since there is no current routine method for the capture and handling of these large animals and there is presently no practical method to obtain blood samples remotely from free-ranging whales. Therefore, we here not only intend to review the endo- and ectoparasite fauna of large whales but also to provide new insights in current available methods for gathering parasitological data by using non- or minimally invasive sampling techniques. We focus on methods, which will allow detailed parasitological studies to gain a broader knowledge on parasitoses affecting wild, free-swimming large whale populations.

Parasite communities and feeding ecology of the European sprat (Sprattus sprattus L.) over its range of distribution.

The metazoan parasite fauna and feeding ecology of 165 Sprattus sprattus (L., 1758) was studied from different geographic regions (Baltic Sea, North Sea, English Channel, Bay of Biscay, Mediterranean Sea). A total of 13 metazoan parasite species were identified including six Digenea, one Monogenea, two Cestoda, two Nematoda and two Crustacea. Didymozoidae indet., Lecithocladium excisum and Bomolochidae indet. represent new host records. The parasite species richness differed according to regions and ranged between 3 and 10. The most species-rich parasite fauna was recorded for sprats from the Bay of Biscay (North Atlantic), and the fishes from the Baltic Sea contained the lowest number of parasite species. More closely connected geographical regions, the North Sea, English Channel and Bay of Biscay, showed more similar parasite component communities compared with more distant regions. From the examined stomachs of S. sprattus, a total of 11 different prey items were identified, including Mollusca, Annelida, Crustacea and Tunicata. The highest number of prey organisms belonged to the crustaceans. The variety of prey items in the stomach was reflected by the parasite community differences and parasite species richness from the different regions. The feeding ecology of the fish at the sampled localities was responsible for the observed parasite composition and, secondarily, the zoogeographical distribution of the parasites, questioning the use of the recorded sprat parasites as biological indicators for environmental conditions and change.

Genetic variability in Hysterothylacium aduncum, a raphidascarid nematode isolated from sprat (Sprattus sprattus) of different geographical areas of the northeastern Atlantic.

Species of the genus Hysterothylacium are among the most common marine nematode fish parasites in the northern Atlantic. Due to recent findings of cryptic speciation in other parasitic ascaridoid nematodes, a similar pattern of sibling species was hypothesized also for Hysterothylacium aduncum. By investigating a 886- to 890-bp-long genomic DNA fragment including ITS-1, 5.8S rDNA and ITS-2 of 40 specimens of H. aduncum of sprat (Sprattus sprattus) of four different biogeographical regions (North Sea, English Channel, Bay of Biscay, Adriatic Sea), we could not detect significant genetic variability and therefore cryptic speciation. Nevertheless, while ITS-1 and 5.8S rDNA sequences were identical for all analysed specimens, ITS-2 sequences showed a population-specific pattern with the differentiation of an English Channel/Bay of Biscay group from a North Sea/Mediterranean Sea group.