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.

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.

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.

Re-description and molecular characterisation of Choricotyle australiensis Roubal, Armitage & Rohde, 1983 (Monogenea: Diclidophoridae) infecting Chrysophrys auratus (Forster) (Perciformes: Sparidae).

Choricotyle australiensis Roubal, Armitage & Rohde, 1983, a diclidophorid monogenean species, is redescribed and genetically characterised using the partial nuclear 28S ribosomal RNA gene (28S rRNA) and a fragment of mitochondrial cytochrome c oxidase subunit 1 (cox1) gene sequences for specimens collected from Chrysophrys auratus (Forster) off Australia and New Zealand. Previous studies have either provided morphological or genetic results, whereas this study combines morphological and advanced molecular methods. A total of 70 Ch. auratus were examined with 22 individuals of C. australiensis recovered from the gills (overall prevalence of 23%). This study has provided the first evidence for the exploration of mitochondrial cox1 region for C. australiensis. Comparison of the newly generated sequences with other available data supported the distinction of C. australiensis among diclidophorid Furhmann, 1928 species thus confirming its taxonomic status.

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.

Genetic characterisation of Tanqua (von Linstow, 1879) (Nematoda: Gnathostomatidae) larval forms including new host and locality records.

In an unrelated study of spotted snakehead fish Channa punctata (Bloch) of family Channidae (N = 103) from Bangladesh, ten fish had taupe and clear coloured cysts attached to the intestinal mesentery. Investigation of the cysts revealed larval nematodes. The larvae were damaged and not suitable for detailed morphological study, however, key features such as tooth like projections of the pseudolabia and lateral pseudolabium were observed in specimens with undamaged cephalic regions. Molecular characterisation was undertaken and although the parasite genetic material was poor, five of the twelve nematode larvae through sequencing of the 18S ribosomal RNA gene, showed 98.17% match with sequences assigned for Tanqua tiara (accession number JF934728) deposited in GenBank. The prevalence of infection was 9.7% and the mean intensity 2.70. Tanqua has not previously been identified in fish, or from the definitive host, the Asian water monitor Varanus salvator (Laurenti, 1768) of family Varanidae (class Reptilia), in Bangladesh. Therefore, this study represents a new host and locality record for this nematode species. In many previous reports from this region, nematode larvae have been identified morphologically and assigned to a diverse range of nematode genera. Some confusion therefore exists regarding their accuracy and further investigations are required using molecular methodology to clarify the species of larval nematodes which infect edible fish in Bangladesh.

Molecular characterisation of the monogenea parasites of blue mackerel Scomber australasicus (Perciformes: Scombridae) in Australian waters.

This study describes the occurrence and molecular identification of Monogenea from blue mackerel Scomber australasicus (Cuvier) (Perciformes: Scombridae), an edible fish, from Australian waters. Previous studies have provided either morphological or genetic results, whereas this study combines both methods of species identification. A total of 50 fish sourced from the waters off the south-eastern Australian coastline were examined and 71 Monogenea were recovered from the gills. The overall prevalence, mean intensity, and mean abundance were 64%, 2.22, and 1.42, respectively. Monogenea were initially classified morphologically as five species belonging to two families. Family Mazocraeidae was represented by Kuhnia scombri (Kuhn, 1829) Sproston, 1945, K. scombercolias Nasir & Fuentes Zambrano, 1983 and Pseudokuhnia minor (Goto, 1894) Rohde & Watson, 1985 and family Gastrocotylidae by Gastrocotyle kurra Unnithan, 1968 and Allogastrocotyle bivaginalis Nasir & Fuentes Zambrano, 1983. Molecular identification of Monogenea was conducted through sequencing of the mitochondrial cytochrome c oxidase subunit 1 (cox1) gene. The host S. australasicus was barcoded (cox1) to confirm the specific identity. There was no comparable sequence available in GenBank for K. scombercolias. Also, limited sequences were available in GenBank for the gastrocotylid Monogenea identified in this study. However, phylogenetic analyses of cox1 sequences of the Monogenea identified in this study clustered according to their familial groups. Gastrocotyle kurra and A. bivaginalis were identified for the first-time on S. australasicus in Australian waters. This study provides the first sequencing of cox1 gene for K. scombercolias. The outcomes of the study provide a basis for future Monogenea research in Australian waters, as well as for other Scomber spp.

Occurrence and abundance of zoonotic nematodes in snapper Chrysophrys auratus, a popular table fish from Australian and New Zealand waters.

In Australia and New Zealand (NZ), snapper Chrysophrys auratus is known for delicate mild flavoured flesh and is a favoured species to serve raw as sashimi or in sushi. The diet of snapper includes a variety of intermediate hosts of larval nematodes, and as a result, snapper has potential to become highly infected with zoonotic/non-zoonotic nematodes. The aims of this study were to survey nematodes in snapper from Australia and New Zealand waters and to identify nematode species using combined morphological and molecular methods. The zoonotic potential of nematodes identified in this study are discussed. A total of 112 snapper were purchased from the Sydney fish market, New South Wales, Australia. Fish were dissected and only the visceral content and digestive tract were examined for nematode infection. Parasites were initially identified by the microscopic method as four different types belonging to the families Anisakidae (Anisakis types I & III, and Terranova type II) and Cucullanidae (Dichelyne spp.). All Anisakidae nematodes were at infective stages. Species-level identification was actualised through sequencing of the internal transcribed spacer (ITS-1, 5.8S, ITS-2) regions. The Anisakis types I & III were confirmed as Anisakis pegreffii and A. brevispiculata, respectively of which A. pegreffii is considered globally as a zoonotic nematode. The specific identification of Terranova type II and Dichelyne spp. was not possible as no comparable sequence data were available in GenBank. The phylogenetic tree clustered Anisakis types I & III with A. pegreffii and A. brevispiculata, respectively; Terranova type II sequences as a separate clade with previously identified larval and adult Terranova and Pseudoterranova species. Based on phylogenetic analyses the present Cucullanid specimens were assigned herein as Dichelyne cf. pleuronectidis, and an unknown species Dichelyne sp. 1. This study represents the first host record globally for zoonotic Anisakid nematodes in this popularly consumed table fish and a new region record for D. cf. pleuronectidis and Dichelyne sp. 1. Further investigation is required, using more comprehensive parasite detection and recovery methods, to assess the health risk these nematodes may pose to human and fish health in Australia/NZ.

Zoonotic nematode parasites infecting selected edible fish in New South Wales, Australia.

Despite increases in the annual consumption of seafood in Australia, studies on the occurrence and prevalence of zoonotic parasites in fish and the risk they may pose to human health are limited. The present study was aimed at determining the occurrence of zoonotic nematodes in commonly consumed fish in New South Wales, Australia's most populous state. Three species of fish, including the Australian pilchard, Australian anchovy, and eastern school whiting, were purchased from a fish market and examined for the presence of nematode parasites. All Australian pilchards examined in this study were infected (100%; n = 19), followed by the eastern school whiting (70%; n = 20) and Australian anchovy (56%; n = 70). Nematodes were in the larval stage and, therefore, classified by morphotype, followed by specific identification through sequencing of their internal transcribed spacer (ITS) regions. Seven different larval types with zoonotic potential, belonging to the families Anisakidae (Contracaecum type II and Terranova type II) and Raphidascarididae (Hysterothylacium types IV [genotypes A and B], VIII, XIV and a novel Hysterothylacium larval type, herein assigned as type XVIII), were found. The new larval type was identified as Hysterothylacium thalassini, based on ITS sequence data. The presence of the infective stage of a range of zoonotic parasites in fish commonly consumed in New South Wales is important, particularly as, in some dishes, these fish are used whole, raw or undercooked. This study provides the basis for future research on other aspects of these parasites, in regards to public health.