Characterisation of Nematoda and Digenea in selected Australian freshwater snails.

Freshwater snails are integral to local ecosystems as a primary food source for various vertebrate species, thereby contributing significantly to ecological food webs. However, their role as intermediate hosts also makes them pivotal in the transmission of parasites. In Australia, research on freshwater snails has predominantly focused on their role as intermediate hosts for livestock parasites, while there has been limited exploration of the impact of these parasites on snail health and population dynamics. The aim of this study was to determine parasitic infection in freshwater snails. This study was conducted in the south-eastern region of Australia, in 2022. A total of 163 freshwater snails from four different species were collected and examined in the Murrumbidgee catchment area in the southeastern part of Australia during the Southern Hemisphere summer and autumn months (February to May). The species included Isidorella hainesii, Glyptophysa novaehollandica, Bullastra lessoni (endemic species), and Physella acuta (an introduced species). Through the analysis of sequence data from the various regions of the nuclear ribosomal DNA, we determined that the Digenea species in this study belonged to three distinct species, including Choanocotyle hobbsi, Petasiger sp. and an unidentified species belonging to Plagiorchioidea. Additionally, analysis of the sequences from Nematoda found in this study, revealed they could be categorized into two separate taxa, including Krefftascaris sp. and an unidentified nematode closely associated with plant and soil nematodes. This research holds significant implications for the future understanding and conservation of Australian freshwater ecosystems. Most parasites found in the present study complete their life cycle in snails and turtles. As many of freshwater snail and turtle species in Australia are endemic and face population threats, exploring the potential adverse impacts of parasitic infections on snail and turtle health, is crucial for advancing our understanding of these ecosystems and also paving the way for future research and conservation efforts. While none of the native snail species in the present study have been listed as endangered or threatened, this may simply be attributed to the absence of regular population surveys.

Occurrence of Stenurus globicephalae (Nematoda: Pseudaliidae) in the blowhole of Globicephala macrorhynchus (Cetacea: Delphinidae) in Tasmania, Australia.

Information about parasites of cetaceans in Australia is scarce and mostly opportunistic. The morphology of specimens of the metastrongyloid Stenurus globicephalae Baylis & Daubney, 1925 (Nematoda: Pseudaliidae), collected from the blowhole of a pilot whale Globicephala macrorhynchus Gray, 1846 (Cetacea: Delphinidae) off northern Tasmania, Australia, were studied. Light and scanning electron microscopical examinations enabled a detailed redescription of this nematode species, including corrections of some inaccuracies in previous species descriptions, particularly those concerning cephalic and caudal structures. The presence of numerous ventrolateral oblique muscle bands, characteristic of the males of S. globicephalae, is reported for the first time. This is the second finding of this nematode parasite, in a different host species, in Tasmania.

Multidisciplinary approach to the diagnosis of Contracaecum magnipapillatum infections in Australian black noddies, Anous minutus (Charadriiformes: Laridae).

We provide the incidental necropsy findings associated with anisakid nematode infections of black noddy terns, Anous minutus Boie, 1844 (Charadriiformes: Laridae), from offshore islands in the southern Great Barrier Reef, Queensland, Australia. Specimens collected from the proventriculi were identified morphologically as Contracaecum magnipapillatum Chapin, 1925 (Rhabditida: Anisakidae), using light and scanning electron microscopy (SEM). The entire nuclear ribosomal DNA internal transcribed spacer (ITS) region (ITS1-5.8S-ITS2) was amplified by polymerase chain reaction (PCR) and sequenced to provide reference sequences for morphologically well-identified voucher specimens. Interestingly, after an alignment with closely related taxa using BLAST, sequences of the ITS1 and ITS2 were 100% identical to the sequences assigned to Contracaecum septentrionale Kreis, 1955, from a razorbill, Alca torda Linnaeus, 1758 (Charadriiformes: Alcidae), from Spain. These results either raise questions about the ITS as a genetic marker for some members of Contracaecum, or the identity of the specimens assigned to C. septentrionale, given that no supporting morphological data was associated with them. We highlight the need for a combined morphological and molecular approach to parasite diagnostics and the use of multiple genetic loci to resolve the molecular taxonomy of cryptic species. Morphological identifications should be taxonomically robust, transparent and precede the deposition of molecular barcodes in public repositories. The gross and histopathological findings of our investigation concur with previous reports of widespread Contracaecum infections in black noddies and support the contention that Contracaecum spp. are an unlikely primary cause of mortality.

A critical review of anisakidosis cases occurring globally.

A review was conducted to identify the most common causative agents of anisakidosis, the methods used for identification of the causative agents, and to summarize the sources of infection, and patients' demographics. A total of 762 cases (409 articles, inclusive of all languages) were found between 1965 and 2022. The age range was 7 months to 85 years old. Out of the 34 countries, Japan, Spain, and South Korea stood out with the highest number of published human cases of anisakidosis, respectively. This raises the question: Why are there few to no reports of anisakidosis cases in other countries, such as Indonesia and Vietnam, where seafood consumption is notably high? Other than the gastrointestinal tract, parasites were frequently found in internal organs such as liver, spleen, pancreas, lung, hiatal and epigastric hernia, and tonsils. There are also reports of the worm being excreted through the nose, rectum, and mouth. Symptoms included sore throat, tumor, bleeding, gastric/epigastric/abdominal/substernal/lower back/testicular pain, nausea, anorexia, vomiting, diarrhea, constipation, intestinal obstruction, intussusception, blood in feces, hematochezia, anemia, and respiratory arrest. These appeared either immediately or up to 2 months after consuming raw/undercooked seafood and lasting up to 10 years. Anisakidosis commonly mimicked symptoms of cancer, pancreatitis, type I/II Kounis syndrome, intussusception, Crohn's disease, ovarian cysts, intestinal endometriosis, epigastralgia, gastritis, gastroesophageal reflux disease, hernia, intestinal obstruction, peritonitis, and appendicitis. In these cases, it was only after surgery that it was found these symptoms/conditions were caused by anisakids. A range of not only mainly marine but also freshwater fish/shellfish were reported as source of infection. There were several reports of infection with >1 nematode (up to >200), more than one species of anisakids in the same patient, and the presence of L4/adult nematodes. The severity of symptoms did not relate to the number of parasites. The number of anisakidosis cases is grossly underestimated globally. Using erroneous taxonomic terms, assumptions, and identifying the parasite as Anisakis (based solely on the Y-shaped lateral cord in crossed section of the parasite) are still common. The Y-shaped lateral cord is not unique to Anisakis spp. Acquiring a history of ingesting raw/undercooked fish/seafood can be a clue to the diagnosis of the condition. This review emphasizes the following key points: insufficient awareness of fish parasites among medical professionals, seafood handlers, and policy makers; limited availability of effective diagnostic methodologies; and inadequate clinical information for optimizing the management of anisakidosis in numerous regions worldwide.

Genetic characterization and phylogenetic relationships of Phyllodistomum parasites in Indian subcontinent: insights from freshwater fish and shrimp hosts.

Phyllodistomum is the large digenean group of fish parasites, with 25 species described so far in the Indian subcontinent. Here, we redescribed two adult species of Phyllodistomum (P. srivastava Rai 1964 and P. parorchium Jaiswal 1957) collected from freshwater fish Heteropneustes fossilis Bloch, 1974 and Glossogobius giuris Ham, 1822, respectively, and an unknown Phyllodistomum metacercaria from shrimp (Macrobrachium dayanum Henderson, 1893). These parasites were genetically characterized using 28S and first and second internal transcribed spacers (ITS1 and ITS2) regions of the nuclear ribosomal DNA and CoxI region of the mitochondrial (mt) DNA to establish the link between metacercaria and adult. Morphologically, both the unknown metacercaria in shrimp and adult Phyllodistomum srivastava in fish, resembled in terms of crenulated margin of hind body, arrangement of diagonal testes, bipartite seminal vesicle, and compact paired vitelline masses. The two adult parasite species, P. srivastava from P. parorchium, were different in terms of shape and size of the body, ratio of suckers, the absence of crenulated margin of hind body, a single chambered seminal vesicle, and deeply lobed paired vitelline masses, in the former species. Comparison of the 28S, ITS, and mtCoxI sequence data suggested P. srivastava and Phyllodistomum metacercaria belong to the same species, and supported the distinction between P. srivastava and P. parorchium. Exploring the potential impact of Phyllodistomum infection on host behaviour and health would be prospective areas for future research.

Characterization of Clinostomum (Digenea: Clinostomidae) spp. in India.

Platyhelminths belonging to the family Clinostomidae (Digenea) have a worldwide distribution and are known to infect piscivorous birds through their intermediate hosts, usually fish species. In the present study, clinostome metacercariae were collected from fish hosts, including Channa punctata (Bloch 1793) (n = 25) and Trichogaster fasciata Bloch and Schneider 1801 (n = 25), from a freshwater system in India. The experimental infection of cattle egrets, Ardea (Bubulcus) ibis Linnaeus 1758, with some of the live metacercariae found in the present study was successful. Live adult parasites were obtained from the buccal cavity of the birds. Both metacercaria and adult specimens were subjected to molecular studies to obtain the sequences of 28S, ITS1, and ITS2 (nuclear rDNA) regions. The parasites were found to belong to three species, Clinostomum giganticum Agarwal 1959; C. piscidium Southwell and Prashad 1918; and Euclinostomum heterostomum (Rudolphi 1809). Phylogenetic analyses of the sequences obtained from the adults and metacercariae established a link between the metacercariae in the fish hosts and adults in the avian host, which is essential to elucidate their partial life cycle and specify morphological characteristics in the metacercarial stage.

Characterization of tongue worms, Linguatula spp. (Pentastomida) in Romania, with the first record of an unknown adult Linguatula from roe deer (Capreolus capreolus Linnaeus).

Specimens of the pentastomid parasite, Linguatula serrata, have been reported from several animals in Romania, including some domestic dogs translocated to other parts of Europe. In this study, gray wolves (Canis lupus, n = 80), golden jackals (C. aureus, n = 115), red foxes (Vulpes vulpes, n = 236), and roe deer (Capreolus capreolus, n = 1) were examined for pentastomes. Overall, 17.5% of wolves were found to be infected with specimens of Linguatula, with a range of infections of one to five individuals per animal. Golden jackals and foxes had much lower infection levels, with 1.73% of golden jackals and 1.69% of foxes infected; both host species were found to be infected with one or two individual pentastomes per animal. The single deer specimen was infected with three individual pentastomes. The pentastomes collected from the wolves and golden jackals were determined to be immature and mature adult specimens of L. serrata based on morphological examination and molecular analysis using the 18S rRNA gene. No pentastomes collected from the red foxes were available for identification. The pentastomes collected from the roe deer were expected to be L. arctica but determined to be mature adult male specimens of an unknown Linguatula, herein, referred to as Linguatula sp. based on its morphology; the results of molecular sequencing for the Linguatula specimen collected from the deer were inconclusive, preventing a final species identification. This study presents the first report of L. serrata in any hosts from Romania through both morphological and molecular characterization, and also presents the first report of a Linguatula sp. in Ca. capreolus, utilizing morphological characterization. Issues of morphological variability are discussed, including the presence of spines in the hook pit of specimens of Linguatula. This study highlights the need to examine all specimens of Linguatula to confirm the stage of development. Despite the inconclusive molecular result for some specimens, the authors still urge future researchers to incorporate a combined molecular and morphological approach in identifying specimens of Linguatula.

Occurrence and molecular identification of Monogenea from blue-spotted flathead Platycephalus caeruleopunctatus (Scorpaeniformes: Platycephalidae) in Australian waters.

This study describes the occurrence and molecular identification of Monogenea from blue-spotted flathead Platycephalus caeruleopunctatus (McCulloch) (Scorpaeniformes: Platycephalidae) from waters off the NSW coast, Australia. Platycephalus spp. are favoured by consumers for delicate, white, mild flavoured flesh and therefore are commercially important species within Australia. Platycephalus spp. are also extensively targeted by Australian recreational fishers. There has been no previous study that has exclusively focused on Pl. caeruleopunctatus in Australia or globally. Although a single study by Dillon (1985), of monogeneans infecting Platycephalus spp. from Australian waters, identified Microcotyle bassensis Murray, 1931 in Pl. caeruleopunctatus. The present study combines both morphological and molecular methods to identify both host and parasites. A total of 116 fish, sourced from the waters off the coast of New South Wales, Australia, were examined. A total of 1498 Monogenea were recovered from the gills. The overall prevalence, mean intensity, and mean abundance were 72%, 18.05, and 12.91, respectively. Monogenea were initially classified morphologically as two different species M. bassensis (family: Microcotylidae) and Platycephalotrema bassense (Hughes, 1928) Kritsky & Nitta, 2019 (family: Ancyrocephalidae). Molecular identification of Monogenea was conducted through sequencing of their mitochondrial cytochrome c oxidase subunit 1 (cox1) and nuclear 28S genes. The specific identification of host Pl. caeruleopunctatus was confirmed through sequencing the cox1 gene. There was no comparable sequence for cox1 and 28S genes available in GenBank for the monogenean species found in the present study. Only a single sequence (obtained from the nuclear ITS2-rDNA) was deposited in GenBank for M. bassensis. However, the phylogenetic analyses of mitochondrial and nuclear sequences revealed that the identified Monogenea clustered according to their familial groups. Platycephalotrema bassense was identified for the first time in Pl. caeruleopunctatus in the present study. This study has provided the first evidence for the exploration of both cox1 and 28S sequences of all Monogenea. The findings of this study serve as a foundation for future monogenean research on other Platycephalus spp. from Australian waters.

28S rRNA sequences for Linguatula spp.

Identification of specimens belonging to the genus Linguatula (Pentastomida) is relatively easy due to their unique morphology. However, differentiation between species of Linguatula can be challenging for several reasons, including considerable differences between different developmental stages of the parasite within and between species. Currently, 18S rRNA and Cox1 sequences are the only available comparable sequences in GenBank, but recent research has discussed the utility of 28S rRNA for pentastomid phylogenetics. This study presents 28S rRNA gene sequences for two members of the genus Linguatula. Sequences of 28S rRNA were successfully obtained from well-identified samples of L. serrata (collected in Australia) and L. nuttalli (collected in South Africa), with voucher specimens. Phylogenetic analysis of the 28S rRNA region showed 6% difference between L. serrata and L. nuttalli, with low levels of intraspecific variation. In comparison, 18S rRNA and Cox1 sequences from the same specimens showed 0.23% and 13% interspecific differences, respectively. The results of this study show that 28S rRNA has greater genetic diversity to allow for improved differentiation between species of Linguatula than 18S rRNA but is on par with Cox1. Records that do not provide adequate morphological or molecular data to justify independent specific diagnoses must be regarded cautiously, and the need for continued research on species of Linguatula, using a combined morphological and molecular analysis, across a number of different hosts, development stages, geographical regions and molecular markers is highlighted.

Molecular (cox1), geographical, and host record investigation of monogeneans Mazocraes australis (Mazocraeidae), Polylabris sillaginae, and P. australiensis (Microcotylidae).

This study determines the occurrence and molecular characterisation of Monogenea from three commercially important Australian fish: Australian sardine Sardinops sagax (Jenyns), Australian anchovy Engraulis australis (White), and eastern school whiting Sillago flindersi McKay. Earlier studies have provided only morphological species identification, whereas this study combines both morphological and molecular methods. A total of 247 fish across 3 species, sourced from the New South Wales and Victorian coasts, were examined for Monogenea. A total of 187 monogenean parasites were recovered from the gills. The overall prevalence, mean intensity, and mean abundance were 34%, 2.23, and 0.78, respectively. The parasites were initially classified morphologically as three species across two families. Family Mazocraeidae was represented by Mazocraes australis Timi et al. J Parasitol 85:28-32, 1999, and family Microcotylidae by Polylabris sillaginae (Woolcock, Parasitology 28:79-91, 1936) Dillon, Hargis, and Harrises, 1983 and P. australiensis Hayward, 1996. Molecular identification of parasites was conducted through sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The fish hosts in the present study were also barcoded (mitochondrial cox1 gene) to confirm specific identities. There was no comparable cox1 sequence available in GenBank for the parasites found in the present study. However, the phylogenetic tree clustered the monogenean species identified in this study according to their familial groups of Mazocraeidae and Microcotylidae. The presence of M. australis on E. australis and S. sagax was confirmed in this study. Polylabris australiensis was only found on S. sagax but Si. flindersi was found to be a host for both Polylabris species. This study is the first to explore the mitochondrial cox1 genes of these three-monogenean species. These findings will serve as a foundation for future monogenean research in Australian waters and elsewhere.

Phylogenetic relationships of philometrid nematodes (Philometridae Baylis & Daubney, 1926) inferred from 18S rRNA, with molecular characterisation of recently described species.

Nematodes of the family Philometridae Baylis & Daubney, 1926 (Dracunculoidea Stiles, 1907) are generally poorly known, and there are many taxonomic issues within the family. Philometrids are parasites of fish and are found in various locations throughout the host, including within the subcutaneous tissues and musculature, the abdominal cavity and gonads; vast sexual dimorphism often means the males are not collected, leading to many species being described solely on female characteristics. Although there have been a number of studies utilising molecular data, the vast majority of species are yet to be sequenced. This study undertook genetic sequencing of 15 recently described species of philometrids across 4 genera, many of which were from specimens collected from waters off Australia. All of the sequences obtained were closely related with representatives of the family Philometridae. Species were found to be distributed in the phylogenetic trees within 4 clades based on a combination of site of infection within the host and host habitat. Family of host and geographical location was not as important for position within the trees. Clade A contained philometrids collected from the abdominal cavities and head tissues of South American freshwater fish. Clade B contained philometrids primarily from the abdominal cavities of freshwater European cyprinids. Clade C contained philometrids primarily from the ovaries of marine fish. Clade D contained philometrids from the body tissues of marine and freshwater fish. The potential co-evolutionary patterns between philometrids and their fish hosts are highlighted as an area of future research. This research also highlighted the importance of correct identification of any sequenced specimen.

Dolichoperoides macalpini (Nicoll, 1914) (Digenea: Dolichoperoididae) infecting venomous snakes (Elapidae) across Australia: molecular characterisation and infection parameters.

Specimens of Dolichoperoides macalpini (Nicoll, 1914) (Digenea: Dolichoperoididae) were collected from Australian venomous snakes (Elapidae): Notechis scutatus Peters, 1861 and Austrelaps superbus (Günther, 1858) from Tasmania and surrounding islands and N. s. occidentalis Glauert, 1948 from wetlands near Perth, Western Australia. Despite variation in morphological measurements, genetic analysis showed that the one species of digeneans infected the snakes from all locations. This study presents the first DNA sequences for D. macalpini (internal transcribed spacer, 18S, 28S), confirming its placement in a family separate from the Reniferidae and Telorchiidae. Analysis of the infection dynamics of infection in Western Australian snakes showed significant differences in levels of infection between wetland locations, season and year of collection. Infection of D. macalpini was reported in the gastrointestinal tract, including the mouth, in freshly euthanised snakes in Western Australia, and in the lung in Tasmanian snakes, consistent with earlier reports. Differences in morphology and site of infection are suggested to be due to a combination of season and maturity of the digenean, with infection potentially occurring early in the season, as the snakes emerge from torpor. The need for research on the seasonal dynamics of infection with this parasite is discussed.

The occurrence and abundance of infective stages of zoonotic nematodes in selected edible fish sold in Australian fish markets.

Seafood is nutritious and a healthy source of proteins and its regular consumption is highly recommended by medical professionals and dieticians. Owing to this, the global consumption of seafood per capita has been significantly increasing since the 1960s. Consequently, seafood-borne pathogens, including parasites, have also become more widely known and recognised. In Australia, a vast island country, information about such parasites is extremely limited. This study aimed to determine the prevalence and abundance of zoonotic parasites, including anisakid nematodes, in selected Australian edible fish. Four species of fish, namely tiger flathead, Platycephalus richardsoni (n = 43), blue mackerel, Scomber australasicus (n = 117), snapper, Pagrus auratus (n = 11) and school whiting, Sillago flindersi (n = 90) were purchased from a fish market. Although a range of parasites was found, due to their significance for human health, the focus of this study was on nematodes whose infectious stage was found in these fish. The prevalence of nematodes in these fish species was 86.05%, 64.10%, 45.45% and 56.67%, respectively. Among the parasites found, Anisakis spp., Contracaecum spp. and Hysterothylacium spp. in tiger flathead, blue mackerel and school whiting, might be of zoonotic importance. Our findings suggest there is a need to revise current seafood safety protocols and develop educational campaigns for seafood industries stakeholders.

The taxonomic position of Anoplotaenia dasyuri (Cestoda) as inferred from molecular sequences.

Anoplotaenia dasyuri Beddard, 1911 (Cestoda), from the Tasmanian devil, Sarcophilus harrisii (Boitard, 1842), is a taxonomic enigma, where a combination of morphological features, host type and geographical location have prevented it from being placed within a family and it is considered incertae sedis, despite its accepted validity. We performed a phylogenetic analysis of three A. dasyuri specimens collected from three Tasmanian devils using 18S and 28S rRNA sequences. Anoplotaenia dasyuri was found to have closest affinity with the family Paruterinidae, especially the genus Cladotaenia Cohn, 1901. The postulated theory of transfer of an ancestor of Anoplotaenia Beddard, 1911 transferring to the Tasmanian devil from an unrelated carnivorous host, such as an accipitriform or other carnivorous bird, is discussed and supported.

The occurrence of Anisakis spp. in Australian waters: past, present, and future trends.

As one of the world's megadiverse countries, Australian biodiversity is vital for global biodiversity. Nematodes belonging to the genus Anisakis (family Anisakidae) are an important part of this biodiversity due to their ability to be repeatedly transmitted among their intermediate hosts before reaching the top of the food pyramid. Therefore, they have a significant impact on the community structures of various ecosystems. In addition, globally, they are known to be of medical and veterinary significance. The aim of this article is to provide an update on the current knowledge about these important parasites in Australia. Since 1916, a total of 234 records of Anisakis spp. from various hosts and localities have been found in Australia. It is estimated that the occurrence of Anisakis spp. and their health impacts in at least 84, 98.5, and 95% of Australian marine mammals, fish, and water birds, respectively, have not been documented yet. The results of this study suggest Australia is perhaps home to the most diverse Anisakis fauna. Available information is dominated by reports of these parasites in fish hosts, many of them among edible fish. Given the popularity of seafood in Australia and the occurrence of infectious stages of Anisakis spp. in edible fish, all stakeholders should be made aware of the occurrence, prevalence, and survival of Anisakis spp. in seafood. Also, as more pet owners feed their pets with a variety of fish and seafood products, it is important for veterinarians to be aware of seafood transmitted Anisakis spp. in pet animals. This study also highlights several important knowledge gaps: (i) The detailed life cycle of Anisakis spp. in Australia is not known. Detecting their first intermediate hosts is important for better management of crustacean zooplankton populations in our waters. (ii) Research on Anisakis spp. in Australia has been restricted to limited taxonomical studies and should extend to other aspects of these important parasites. (iii) The capacity to identify parasite taxa to species is especially important for resolving biological diversity around Australia; however, opportunities to formally train in parasite taxonomy are rare and diminishing. There is a need to train researchers with taxonomy skills. (iv) Given the vast range of biodiversity in Australia and the broad host-specificity of Anisakis spp., particularly in the larval stages, the full range of their intermediate hosts remains unknown. (v) The health impacts of the infection of the intermediate/definitive hosts with Anisakis spp. are not fully understood. Thus, one of the important areas for future studies is investigating the pathogenicity of Anisakis spp. in affected animals. This is a crucial yet unknown factor for the conservation of some endangered species in Australia.

Integrative species delimitation and community structure of nematodes in three species of Australian flathead fishes (Scorpaeniformes: Platycephalidae).

This study aimed to determine the integrative characterisation of nematodes from three species of edible flathead fishes (Scorpaeniformes: Platycephalidae) in New South Wales, Australia, and describe nematode communities within three species of flatheads. Tiger (Platycephalus richardsoni (Castelnau); n = 20) and sand flatheads (Platycephalus bassensis (Cuvier); n = 20), sourced from the Nelson Bay area, and dusky flathead (Platycephalus fuscus (Cuvier); n = 20) from the Manning River, Taree, were examined for the presence of nematodes. The nematodes were initially classified morphologically as 12 different morphotypes belonging to the families Anisakidae (Anisakis types I, II, and III, Contracaecum type II, Terranova types I and II), Raphidascarididae (Hysterothylacium types IV, VI, VIII, and H. zhoushanense larva), and Gnathostomatidae (Echinocephalus sp. larva), Capillariidae (Capillaria sp.), followed by genetic identification through sequencing of the internal transcribed spacer (ITS-1, 5.8S, ITS-2) regions. Phylogenetic analyses revealed the evolutionary relationship between the identified larval specimens in the present study with available GenBank larval and adult nematodes. Sand flathead was 90% infected with nematodes followed by tiger flathead at 85% and dusky flathead at 15%. Nematodes infecting estuarine dusky and oceanic sand and tiger flatheads contrasted markedly. The analysis of similarities (ANOSIM) showed significant differences (p < 0.001) in the composition of taxa within nematode communities between the three species of flatheads (global R = 0.208) with the highest difference being between sand and dusky flatheads (R = 0.308, p < 0.001). The findings of the present study provide a foundation for future investigations of the community composition, life cycles, and distribution of nematode populations in edible fish in Australia and explore and clarify their significance to public health.

Do parasites influence behavioural traits of wild and hatchery-reared Murray cod, Maccullochella peelii?

This study aimed to investigate the links between parasites and behavioural traits of juvenile Murray cod (Maccullochella peelii). The Murray cod is an endangered Australian freshwater fish for which restocking programs are in place and there is a growing human consumption market. However, little is known about the parasites of these fish and how these parasites influence their behaviour and survival. Fingerlings and yearling fish were sourced from a hatchery and the wild, and after acclimatisation in the laboratory, variation in behavioural traits was examined using emergence, exploration and predator inspection tests. The fish were then euthanised to determine their age and examined for infection with parasites. Wild fish had more camallanid nematodes and lernaeid copepods than hatchery fish. An information theoretic approach using Akaike's Information Criterion (AIC) indicated that infection with protozoan cysts was an important factor for predicting the latency to emerge and explore a new environment, which was interpreted as reduced "boldness". In contrast, the presence of lernaeid copepods was included in two of the four best models predicting predator inspection, indicating that infected fish were less likely to inspect a predator. Source of fish (wild or hatchery) was found to be a strong influence on behavioural responses in all our tests. All parasites found in the present study are known to result in clinical signs of diseases in their fish hosts, raising the possibility that responses in tests of behavioural traits reflect side effects of infection. Additionally, the effect of host adaptation to not show signs of parasite infection, or more simply that the effects on behaviour are subtle and difficult to reveal with small sample sizes, is discussed. Nonetheless, we propose that it is important that infection with parasites is considered in fish behavioural studies both to assess survival behaviour and to avoid misinterpretation of behavioural tests of animal personality.

Characterization of Clinostomum sp. (Trematoda: Clinostomidae) infecting cormorants in south-eastern Australia.

Clinostomum Leidy, 1856 (Trematoda: Clinostomidae) is a cosmopolitan, zoonotic genus of fluke that has been poorly studied in an Australian setting. Following previous reports of reservoir fish in Australian fish ponds being heavily infected with Clinostomum metacercaria, the current study was conducted to determine the specific identity of Clinostomum sp. in inland Australia, by examining and characterizing parasites collected from a potential definitive host, cormorants. A total of 33 parasite specimens belonging to the genus Clinostomum were collected from two cormorants (little black cormorants, Phalacrocorax sulcirostris) that were collected from the Narrandera Fisheries Research Centre, New South Wales, at the same locality where metacercaria of Clinostomum sp. have been reported in fish. All specimens in our study were immature adults. Clinostomum specimens with similar morphology have been identified as C. complanatum in the past, based on their morphological characteristics. However, phylogenetic analyses based on the ITS sequence data in the present study suggest they are the same as the Clinostomum sp. previously reported from carp gudgeons (Hypseleotris spp.) from the same farm, and distinct from C. complanatum. The ITS sequences obtained from the specimens in the present study were most similar to those belonging to C. phalacrocoracis (never reported in Australia). Our specimens formed a distinct clade on the phylogenetic tree and their specific identity awaits until fully mature specimens are described in future studies.

Anisakis allergy: unjustified social alarm versus healthy diet; commentary to the "Letter to the Editor" of Drs Daschner, Levsen, Cipriani, and del Hoyo, referencing to "World-wide prevalence of Anisakis larvae in fish and its relationship to human allergic anisakiasis: a systematic review".

Anisakiasis is an underrecognized condition globally, and accurate diagnosis remains problematic even in countries where the condition is well known. Our "systematic review" was conducted according to Prisma guidelines. The stated basis of our study was "syndromic surveillance." Both methods are recognized in published literature as valid to identify or predict disease and to make accessible large amounts of evidence from published literature. Our study identified Anisakis allergy "hot spots" and other geographical areas where fish are highly infected with Anisakis without commensurate studies of human allergy. Results of our study will open up new lines of enquiry. Norway, used as an example to discredit the scientific integrity of our article, has a cuisine thriving with raw fish dishes and many sushi restaurants. The peer reviewed data sets, confirmed A. simplex sensitization among the Norwegian population, although this has been overlooked by the authors of the "Letter to the Editor." The identification of hot spots in our study may be influential in many ways not the least in raising diagnostic suspicion to expedite accurate diagnosis.

Two species of philometrid nematodes (Philometridae) newly recorded from marine fishes off South Australia, including Philometra inconveniens n. sp. from Hyporhamphus melanochir (Valenciennes) (Hemiramphidae).

Recent examinations of some marine fishes from off the coast of South Australia revealed the presence of two species of Philometra Costa, 1845 (Nematoda: Philometridae): P. inconveniens n. sp. from the ovary (males) and body cavity (subgravid female) of the southern garfish Hyporhamphus melanochir (Valenciennes) (Beloniformes, Hemiramphidae) and Philometra sp. (gravid and subgravid females) from the body cavity of the Australian barracuda Sphyraena novaehollandiae Günther (Perciformes, Sphyraenidae) (new host and geographical records). Specimens of species are described and illustrated based on light and scanning electron microscopical examinations. Philometra inconveniens n. sp. differs from the most similar species P. longa Moravec, Barton & Shamsi, 2021, a parasite of the body cavity of the congeneric host off eastern Australia, mainly by a different structure of the gubernaculum (absence of dorsal barbs and presence of lateral extensions on its distal portion). This indicates a high degree of host specificity of these nematodes in co-occuring congeneric hosts.