Characterization of Echinostoma spp. (Trematoda: Echinostomatidae Looss, 1899) infecting ducks in south-eastern Australia.

Waterbirds, are one of the popular game animals and are of significant relevance to parasite spread due to their ability to fly and migrate great distances in relatively short periods of time. In Australia, however, the knowledge of parasites infecting native waterbirds is lacking with some of the last reports occurring over 50 years ago. The study aimed to characterise Echinostoma spp. infecting wild native Australian ducks found in the southern regions of the Murray Darling Basin (MDB). Ducks (n = 98) were collected from southern New South Wales within the MDB catchment. Three different species of native ducks were found including Anas superciliosa (n = 37), Anas gracilis (n = 47) and Chenonetta jubata (n = 14), of which 4.3 %, 2.7 % and 7.1 %, respectively, were found to be infected with adult stages Echinostoma spp. Examination of the parasites revealed the presence of two morphotypes. The 18S, 28S and ITS rRNA as well as the mitochondrial nad1 genes were sequenced for representative isolates of the two morphotypes. These sequences were then compared with existing sequences of Echinostoma spp. available in the GenBank. Phylogenetic analysis based on the ITS region indicated that the two morphotypes were genetically distinct. Although there are comparable sequences of Echinostoma spp. in Australia these morphotypes appear to be genetically distinct. Based on their distinct morphology and genetics we suggest that these two morphotypes are previously undescribed in Australia. This study sheds light on the presence of Echinostoma parasites in native Australian waterbirds and highlights the need for further research to better understand the diversity and prevalence of these parasites in the region.

First findings of Sarcocystis species in game deer and feral pigs in Australia.

Several wild game meat species, including deer and feral pigs are hunted and consumed in Australia. Feral pigs and deer are not indigenous to Australia, but they have proliferated extensively and established their presence in every state and territory. Following the report of a sambar deer displaying Sarcocystis like white cysts in its rump muscles, the present study was conducted to explore the prevalence of Sarcocystis infections in wild deer and feral pigs in the southeastern regions of Australia. Oesophagus, diaphragm, and heart tissue from 90 deer and eight feral pigs were examined visually for sarcocysts. All results were negative. PCR testing of randomly selected deer and feral pigs yielded positive results, which were subsequently supported by histopathology. This is the first study to report the presence of Sarcocystis spp. in deer and feral pigs in Australia. As no visual cysts were found on the heart or oesophagus that came back positive with PCR, infected animals, particularly those reared free-range, could be passing through meat quality checks unidentified. If people consume this meat without cooking it properly, it may lead to a human infection of Sarcocystis. However, a more targeted study focused on determining the parasite's prevalence and assessing its risks is necessary to determine if it constitutes a food safety issue. As this species has been found not only in feral pigs but also in domestic pigs, the potential for infection spreading between feral pigs and pigs in free-range livestock systems is high, potentially posing a large problem for the Australian pork industry, particularly with the increased emphasis on free-range pig husbandry. Future studies should concentrate on determining the species of Sarcocystis in feral animals commonly consumed as game meat to determine potential zoonotic risks. This could also include a more in-depth look at the prevalence of Sarcocystis infections in other game animals. Identifying where these parasites are present and to what extent, are important areas for future studies.

The first finding of Dictyocaulus cervi and Dictyocaulus skrjabini (Nematoda) in feral fallow deer (Dama dama) in Australia.

Feral deer are widespread throughout Australia with the capacity to impact livestock production via transmission of parasites. Samples of Dama dama (fallow deer), Rusa unicolor (sambar deer), Cervus elaphus (red deer) and an unidentified deer were sourced from various locations in south-eastern Australia for examination for parasites. Adult nematodes were collected from the lungs of all deer species across four separate geographical locations. The nematodes were identified as species of Dictyocaulus through both morphological and molecular means. Species identification based on morphological features was difficult, with many measurements from described species overlapping. Molecular analyses targeting three markers, namely 18S rRNA, ITS2, and cox1 revealed the presence of two distinct species: Dictyocaulus cervi and Dictyocaulus skrjabini. These are the first genetically confirmed reports of species of Dictyocaulus in feral deer in Australia, and although cross-transmission of species of Dictyocaulus with livestock has not yet been reported, it cannot be completely discounted without further research.

Age-Related Changes in Survival Behaviour in Parasite-Free Hatchery-Reared Rainbow Trout (Oncorhynchus mykiss).

Millions of hatchery-reared Rainbow trout are currently released in Australian waters to support recreational fisheries objectives, yet many of these fish die soon after release. In addition, little is known whether these fish harbour parasites that can potentially threaten freshwater ecosystems and human health. Here, we tested the behaviour of hatchery-reared trout using six tank-based tests at six different ages to evaluate their chances of survival and then dissected fish to investigate parasite prevalence. At 7 weeks of age fish readily emerged from a hide and showed the greatest number of startle responses to predators. Behaviour around 25-29 weeks of age was relatively "shy", staying in shelter and avoiding open water. At around 37-41 weeks of age though, behaviour changed, with fish emerging from a hide more readily and exploring the environment. Interestingly, at 58 weeks of age fish were slower to initiate exploration, possibly indicating a return to "shyer" behaviour. All fish underwent thorough parasite examination, revealing no infections. We conclude that knowledge of the behaviour of hatchery-reared fish at different ages is useful for decisions around the timing of release that balance the needs of recreational fishers whilst managing the impact on freshwater ecosystem.

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.

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.

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.

The Presence of a Parasite in the Head Tissues of a Threatened Fish (Bidyanus bidyanus, Terapontidae) from South-Eastern Australia.

The silver perch, Bidyanus bidyanus (Mitchell) (Terapontidae) is a freshwater fish, endemic to the Murray-Darling river system in south-eastern Australia. Population declines have led to the fish being listed as critically endangered by the Australian Government. Knowledge about parasites and diseases of wild populations of freshwater fish are limited in Australia. During an examination of wild-caught silver perch, digenean mesocercaria were observed in the head tissues. A total of five of the 11 silver perch collected from the Wakool River, New South Wales, were infected with mesocercaria. All mesocercaria were found in the head tissues; no mesocercaria were found encysted in the eye lens. The mesocercaria were found to belong to the family Strigeidae based on the sequences of their internal transcribed spacer (ITS) region. The lack of comparable sequences of strigeid digeneans from Australian hosts precludes being able to determine if the mesocercaria found in this study are a new species or representatives of an already described species. However, genetic results confirm that this is a different species to other digeneans previously described from silver perch, thus increasing the number of digeneans reported from B. bidyanus to three species. The presence of digenean mesocercaria in the head tissues of a wild population of silver perch, as found in the present study, is of potential conservation significance. Given the critically endangered conservation status of B. bidyanus, and previous evidence of strigeid infection altering fish behaviour, ecology, and predation mortality, further research on the potential impacts of infection on wild populations is warranted.

Occurrence of digenean parasites in freshwater snails in the Murrumbidgee catchment area, Australia.

Freshwater snails are important hosts in the life cycles of many medically important parasites, particularly for digenetic trematodes such as liver flukes and schistosomes. The current study was conducted to determine the infection of freshwater snails with parasites that can potentially be transmitted to humans within the Murrumbidgee catchment area which is an area of widespread intensive aquaculture in Australia. A total of 116 freshwater snails, belonging to three species (Isidorella hainesii, Glyptophysa novaehollandica and Bullastra lessoni), were examined for the presence of parasites in both man-made and natural environments. The analysis of sequence data, including the internal transcribed spacers (ITS) of nuclear ribosomal DNA, small subunit (18S) ribosomal DNA, and large subunit (28S) ribosomal DNA, indicated that the collected parasites belonged to two distinct genera, namely Clinostomum and Echinostoma. It is noteworthy that species of both of these digenean parasites have the potential to be zoonotic. Cercariae of both Clinostomum and Echinostoma were observed in snails collected from aquaculture settings. It is important to highlight that infectious stages of Clinostomum  has been previously detected in edible fish within Australia. This information raises concerns regarding the potential transmission of these parasites to humans through the consumption of contaminated fish. These findings emphasize the importance of monitoring and controlling the presence of Clinostomum and Echinostoma in aquaculture environments to minimise the risk of zoonotic infections and ensure food safety. Further research and surveillance are needed to better understand the prevalence, transmission dynamics, and potential public health implications associated with these parasites in the context of aquaculture in Australia.

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.

Infection levels of Gnathostomatidae (Nematoda) larvae in commercial fishes in north-eastern Australian waters and related food safety concerns.

The majority of research on the safety of marine edible fish has primarily focused on anisakid nematodes, neglecting the potential risks posed by other parasites, including those belonging to the family Gnathostomatidae. In Australia, there have been few reported cases of human infections with gnathostomatid parasites since 2011. However, due to the absence of a standardized diagnostic test in the country, it is believed that the actual number of infections is higher than reported. This study aimed to assess the occurrence and prevalence of infectious gnathostomatid parasites in selected commercial fish species in Australia. A total of 1947 marine fish from northern Australia, representing 9 families, 16 genera, and 30 species, were examined for gnathostomatid nematode infections. Overall, 12.3 % of the fish were found to be infected with at least one gnathostomatid larva. Among the species examined, the yellow-dabbled flounder (Branchypleura novaezeelandiae) exhibited the highest prevalence (83.3 %; n = 6) and the largest number of gnathostomatid larvae. The identification of the gnathostomatid larvae was confirmed as belonging to the genus Echinocephalus based on both morphological characteristics and sequence data. No significant correlation was observed between the prevalence, mean abundance, and mean intensity of infection with the length or weight of the examined fish species. Notably, several of the infected fish species are considered popular choices in the Australian market. Hence, it is imperative to raise awareness among relevant food safety authorities regarding the occurrence of these parasites. The findings from this study should be taken into consideration for the revision of current seafood safety protocols in the country.

Relationships between local variability in parasite communities of the black-spotted croaker (Protonibea diacanthus) (Teleostei: Sciaenidae) and host population structure and seasonality.

We evaluated spatial and temporal variability in parasite communities from the commercially important tropical marine fish the black-spotted croaker (Protonibea diacanthus) (Teleostei: Sciaenidae) to examine its population structure off the coast of the Northern Territory, Australia. Differences in parasite assemblage between four locations, across three seasons of the year, were used to evaluate the degree of connectivity of the sciaenid across coastal study areas. Analysis of parasite prevalence and mean intensity in these fish suggested the four sampling sites are distinct host populations. Across time, parasite assemblages at the four sites were distinct during the mid-dry (April-August) and build-up (September-November) seasons. During the wet season (December-March) there was substantial overlap in the parasite assemblages at three of the four sites indicating that fish population mixing may be occurring. Parasite assemblages at one nearshore site remained distinct across spatial and temporal scales. Our findings support the utility of parasitic organisms for elucidating the population structure of host species and reiterate the need to account for both spatial and temporal variability when performing stock discrimination analyses.

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.

Protozoan Parasites of Iranian Freshwater Fishes: Review, Composition, Classification, and Modeling Distribution.

This article investigates the occurrence and distribution of parasitic protozoa of Iranian freshwater fishes (both farmed and wild). Our search shows 26 known parasitic protozoan species were recorded from 52 freshwater fish species across different ecoregions of Iran. Most of these fish are edible. While none of the identified protozoan parasites are of zoonotic importance, our study does not exclude presence of zoonotic species in Iranian fishes. Present data suggest the northern and western regions of the country are the main macrohabitat of protozoa (35 parasitic records reported), with the greatest concentration of parasitic protozoa occurring in the Urmia basin in Iran's northwest. The clustered distribution pattern of protozoa among freshwater fish was also more evident in the northern and western parts of the country. The gills and skin were the most infected microhabitats for parasitic protozoa. The highest number of parasites was observed in the fish family Cyprinidae with nine species found in the native fish, Capoeta capoeta. The most diverse host range was observed in the holotrich ciliate, Ichthyophthirius multifiliis isolated from 46 cyprinid species in 39 different locations. However, due to the great richness of fish and extreme habitat diversity, parts of the parasite fauna of Iranian freshwater fish are still poorly understood. Furthermore, current and future changes in climate and environmental parameters, and anthropogenic interventions are likely to affect fish hosts and their parasites.

Description of two new species of Diplectanum Diesing, 1858 (Monogenea: Diplectanidae) collected from Protonibea diacanthus (Lacepède, 1802) (Teleostei: Sciaenidae) from waters off northern Australia.

Two new species of the family Diplectanidae Monticelli, 1903 from the gills of Protonibea diacanthus (Lacepède, 1802) (Teleostei: Sciaenidae) off the northern Australian coast are described. Previous studies have either morphological or genetic results, whereas this study combines morphological and advanced molecular methods to provide the first detailed descriptions for species of Diplectanum Diesing, 1858 from Australia utilising both methodologies. Two new species, Diplectanum timorcanthus n. sp. and Diplectanum diacanthi n. sp., are morphologically described and genetically characterised using the partial nuclear 28S ribosomal RNA gene (28S rRNA) and the internal transcribed spacer 1 (ITS1) partial sequence.

Occurrence and characterisation of Eustrongylides species in Australian native birds and fish.

In Australia, nematodes belonging to the genus Eustrongylides were believed to be endemic species until the late 20th century when they were all considered to be E. excisus, invalid or inquirendae. Although these nematodes have frequently been reported in Australian fish, reptiles, and birds and cause disease or mortality among them, there has been no attempt to date to characterise them genetically. Globally, also, no one has validated or defined suitable genetic markers to distinguish between species of Eustrongylides. In this study, adult Eustrongylides from little black cormorant (Phalacrocorax sulcirostris; n = 3) and larvae from mountain galaxias (Galaxias olidus, n = 2) and a Murray cod (Maccullochella peelii, n = 1), and a Murray cod-trout cod hybrids (Maccullochella peelii x Maccullochella macquariensis, n = 1) were available for morphological examination and molecular characterisation. The adult nematodes from cormorants were identified as E. excisus. Sequences of the 18S and ITS regions were then obtained for all nematodes, which were identical among all specimens (larvae and adults) and also identical to those of E. excisus available in the GenBank. However, only one base pair difference exists between the 18S sequences of E. excisus and E. ignotus, with limited sequences available in GenBank accompanied with proper morphological data for the nematodes. With that limitation in mind, identifying our specimens as E. excisus suggests spill-over - that it is an introduced parasite species that has successfully established its life cycle among Australian native species - may have occurred. Our study is the first report of E. excisus in the little black cormorant, P. sulcirostris. Our results do not exclude the possibility of the occurrence of other species of Eustrongylides, either native or exotic, in Australia. This parasite is zoonotic and with increasing demand for fish and changing dietary preferences, such as the consumption of raw or undercooked fish, its occurrence in the flesh of the fish is concerning. This parasite is also associated with anthropogenic habitat alteration affecting the reproductive success of the infected hosts. Therefore, awareness among the relevant authorities of the presence of the parasite in Australia and its adverse impact on native animals is crucial for the success of conservation plans such as fish recovery and relocation efforts.

Bacteria of Zoonotic Interest Identified on Edible Freshwater Fish Imported to Australia.

Previous research has shown that freshwater edible fish imported into Australia are not compliant with Australian importation guidelines and as a result may be high risk for bacterial contamination. In the present study, the outer surface of imported freshwater fish were swabbed, cultured, confirmatory tests performed and antimicrobial patterns investigated. Channidae fish (Sp. A/n = 66) were contaminated with zoonotic Salmonella sp./Staphylococcus aureus (n = 1/66) and other bacteria implicated in cases of opportunistic human infection, these being Pseudomonas sp. (including P. mendocina and P. pseudoalcaligenes (n = 34/66)); Micrococcus sp. (n = 32/66); Comamonas testosteroni (n = 27/66) and Rhizobium radiobacter (n = 3/66). Pangasiidae fish (Species B/n = 47) were contaminated with zoonotic Vibrio fluvialis (n = 10/47); Salmonella sp. (n = 6/47) and environmental bacteria Micrococcus sp. (n = 3/47). One sample was resistant to all antimicrobials tested and is considered to be Methicillin Resistant S. aureus. Mud, natural diet, or vegetation identified in Sp. A fish/or packaging were significantly associated with the presence of Pseudomonas spp. The study also showed that visibly clean fish (Sp. B) may harbour zoonotic bacteria and that certain types of bacteria are common to fish groups, preparations, and contaminants. Further investigations are required to support the development of appropriate food safety recommendations in Australia.

Occurrence and molecular identification of nematodes from blue mackerel Scomber australasicus Cuvier in Australian waters.

This study determines the occurrence and molecular identification of nematodes from blue mackerel Scomber australasicus Cuvier (Perciformes: Scombridae), an edible fish from Australian waters. A total of 150 fish were sourced from the New South Wales and Victorian coasts. Nematodes were initially classified morphologically as 10 morphotypes belonging to the families Anisakidae (Anisakis morphotypes I and II, Contracaecum morphotype II, Terranova morphotypes I and II), Raphidascarididae (Hysterothylacium morphotypes IV, VI, VIII, and XIV), and Capillariidae (Capillaria sp.), followed by molecular identification through sequencing of their internal transcribed spacer (ITS-1, 5.8S, ITS-2) region. Anisakis morphotype I was confirmed as A. pegreffii Campana-Rouget & Biocca, 1955 and A. berlandi Mattiucci, Cipriani, Webb, Paoletti, Marcer, Bellisario, Gibson & Nascetti, 2014. Anisakis morphotype II and Contracaecum morphotype II were confirmed as A. physeteris Baylis, 1923 and C. ogmorhini Johnston & Mawson, 1941, respectively. Terranova morphotypes I and II were identified as Pulchrascaris australis Shamsi, Barton & Zhu, 2020 and Euterranova pectinolabiata n. comb. (Shamsi, Barton & Zhu, 2019) Moravec & Justine, 2020, respectively. The specific identification of Hysterothylacium morphotypes IV, VI, and VIII was not possible as no comparable adult Hysterothylacium species sequences were available in GenBank, with the exception of morphotype XIV which was confirmed as H. persicum Shamsi, Ghadam, Suthar, Mousavi, Soltani & Mirzargar, 2016. Seven nematode morphotypes were identified for the first time in Australian blue mackerel. The outcomes of the study provide a basis for future research into the community structure, life cycles, and distribution of nematode species in Australian mackerel and to analyse and clarify their importance for public health.

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.

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.