Photobacterium damselae subspecies damselae Pneumonia in Dead, Stranded Bottlenose Dolphin, Eastern Mediterranean Sea.

Photobacterium damselae subspecies damselae, an abundant, generalist marine pathogen, has been reported in various cetaceans worldwide. We report a bottlenose dolphin in the eastern Mediterranean Sea that was found stranded and dead. The dolphin had a severe case of chronic suppurative pneumonia and splenic lymphoid depletion caused by this pathogen.

Multilocus sequence analysis of diverse Streptococcus iniae isolates indicates an underlying genetic basis for phenotypic heterogeneity.

Streptococcus iniae is a Gram-positive, opportunistically zoonotic bacterium infective to a wide variety of farmed and wild fish species worldwide. Outbreaks in wild fish can have detrimental environmental and cultural impacts, and mortality events in aquaculture can result in significant economic losses. As an emerging or re-emerging pathogen of global significance, understanding the coalescing factors contributing to piscine streptococcosis is crucial for developing strategies to control infections. Intraspecific antigenic and genetic variability of S. iniae has made development of autogenous vaccines a challenge, particularly where the diversity of locally endemic S. iniae strains is unknown. This study genetically and phenotypically characterized 11 S. iniae isolates from diseased wild and farmed fish from North America, Central America, and the Caribbean. A multilocus sequence analysis (MLSA) scheme was developed to phylogenetically compare these isolates to 84 other strains of Streptococcus spp. relevant to aquaculture. MLSA generated phylogenies comparable to established genotyping methods, and isolates formed distinct clades related to phenotype and host species. The endothelial Oreochromis mossambicus bulbus arteriosus cell line and whole blood from rainbow trout Oncorhynchus mykiss, Nile tilapia Oreochromis niloticus, and white sturgeon Acipenser transmontanus were used to investigate the persistence and virulence of the 11 isolates using in vitro assays. In vivo challenges using an O. niloticus model were used to evaluate virulence by the intragastric route of infection. Isolates showed significant differences (p < 0.05) in virulence and persistence, with some correlation to genogroup, establishing a basis for further work uncovering genetic factors leading to increased pathogenicity.

Prevalence of nervous necrosis virus (NNV) and Streptococcus species in wild marine fish and crustaceans from the Levantine Basin, Mediterranean Sea.

Infectious diseases in marine animals have ecological, socio-economic and environmental impacts. Nervous necrosis virus (NNV) and Streptococcus iniae have become major threats to marine aquaculture and have been detected in morbid marine organisms worldwide. However, despite their importance, there is a lack of knowledge regarding the prevalence of these pathogens in wild fish species. Here we sampled indigenous and Lessepsian species from different trophic levels and different biological niches in the eastern Mediterranean. A total of 174 fish and 32 crustaceans were tested for S. iniae and a total of 195 fish and 33 crustaceans were tested for NNV. We found an overall prevalence of 9.71% Streptococcus spp. and 21.49% NNV in selected marine fish and crustaceans by PCR and qPCR. In fish, the zoonotic agent S. iniae was detected at a higher prevalence in kidney compared to liver tissue. Co-infection by both pathogens was detected only in 5 specimens. We also examined gilthead sea bream Sparus aurata from an Israeli offshore marine farm during the grow-out period, in order to assess the possibility of horizontal pathogen transmission from wild to maricultured fish. Three out of 15 (20%) fish were found to be NNV positive after 120 d in the sea, suggesting spontaneous transmission from wild to farmed fish. Our findings suggest that more surveys should be conducted, especially in areas were mariculture farms are planned to be established.

Inhibition of nervous necrosis virus by ribavirin in a zebrafish larvae model.

The guanosine analog ribavirin is a broad-spectrum antiviral drug, mostly used in human clinical practice. It has in vitro and in vivo activity against a broad range of RNA and DNA viruses. Here, we report that treatment of zebrafish larvae with ribavirin prior to infection with nervous necrosis virus (NNV) significantly reduces the mortality caused by the virus during the first 10 days post-infection. The RNA genome of NNV harvested from ribavirin-treated infected larvae contains three synonymous and one single non-synonymous mutation, resulting in the replacement of a serine codon with a glycine codon in the RNA-dependent RNA polymerase gene. Adding increasing amounts of guanosine to ribavirin prior to larvae infection did not impede the antiviral activity. Ribavirin treatment of uninfected larvae reduces the basal level of IFNγ, but increases the level of IL-1ß mRNA expression. Furthermore, infecting larvae with NNV following ribavirin treatment reduces the expression levels of IFNγ, IFN-I, Mx, and TNF-α genes, while the expression of IL-1ß is increased. These results suggest that cytokine modulation plays an important role in the activity of ribavirin against NNV. Mortality of more than 40 species of teleost fish, mostly larvae and juveniles, from NNV is a major obstacle in hatcheries, and impedes the supply of young fish to farms. Hence, cost-effective ribavirin treatment should be considered as an efficient means to reduce the peril of NNV.

Detection of Bartonella spp. in wild carnivores, hyraxes, hedgehog and rodents from Israel.

Bartonella infection was explored in wild animals from Israel. Golden jackals (Canis aureus), red foxes (Vulpes vulpes), rock hyraxes (Procavia capensis), southern white-breasted hedgehogs (Erinaceus concolor), social voles (Microtus socialis), Tristram's jirds (Meriones tristrami), Cairo spiny mice (Acomys cahirinus), house mice (Mus musculus) and Indian crested porcupines (Hystrix indica) were sampled and screened by molecular and isolation methods. Bartonella-DNA was detected in 46 animals: 9/70 (13%) golden jackals, 2/11 (18%) red foxes, 3/35 (9%) rock hyraxes, 1/3 (33%) southern white-breasted hedgehogs, 5/57 (9%) Cairo spiny mice, 25/43 (58%) Tristram's jirds and 1/6 (16%) house mice. Bartonella rochalimae and B. rochalimae-like were widespread among jackals, foxes, hyraxes and jirds. This report represents the first detection of this zoonotic Bartonella sp. in rock hyraxes and golden jackals. Moreover, DNA of Bartonella vinsonii subsp. berkhoffii, Bartonella acomydis, Candidatus Bartonella merieuxii and other uncharacterized genotypes were identified. Three different Bartonella strains were isolated from Tristram's jirds, and several genotypes were molecularly detected from these animals. Furthermore, this study reports the first detection of Bartonella infection in a southern hedgehog. Our study indicates that infection with zoonotic and other Bartonella species is widespread among wild animals and stresses their potential threat to public health.

Mortality Caused by Bath Exposure of Zebrafish (Danio rerio) Larvae to Nervous Necrosis Virus Is Limited to the Fourth Day Postfertilization.

Nervous necrosis virus (NNV) is a member of the Betanodavirus genus that causes fatal diseases in over 40 species of fish worldwide. Mortality among NNV-infected fish larvae is almost 100%. In order to elucidate the mechanisms responsible for the susceptibility of fish larvae to NNV, we exposed zebrafish larvae to NNV by bath immersion at 2, 4, 6, and 8 days postfertilization (dpf). Here, we demonstrate that developing zebrafish embryos are resistant to NNV at 2 dpf due to the protection afforded by the egg chorion and, to a lesser extent, by the perivitelline fluid. The zebrafish larvae succumbed to NNV infection during a narrow time window around the 4th dpf, while 6- and 8-day-old larvae were much less sensitive, with mortalities of 24% and 28%, respectively.

Novel evidence suggests that a 'Rickettsia felis-like' organism is an endosymbiont of the desert flea, Xenopsylla ramesis.

Fleas are acknowledged vectors and reservoirs of various bacteria that present a wide range of pathogenicity. In this study, fleas collected from wild rodents from the Negev desert in southern Israel were tested for RickettsiaDNA by targeting the 16S rRNA (rrs) gene. Thirty-eight Xenopsylla ramesis, 91 Synosternus cleopatrae and 15 Leptopsylla flea pools (a total of 568 fleas) were screened. RickettsiaDNA was detected in 100% of the X. ramesis and in one S. cleopatrae flea pools. None of L. algira flea pools was found positive. All positive flea pools were further characterized by sequencing of five additional genetic loci (gltA, ompB, ompA, htrA and fusA). The molecular identification of the positive samples showed all sequences to be closely related to the 'Rickettsia felis-like' organisms (99-100% similarities in the six loci). To further investigate the association between 'R. felis-like' and X. ramesis fleas, ten additional single X. ramesis adult fleas collected from the wild and five laboratory-maintained X. ramesis imago, five larva pools (2-18 larvae per pool) and two egg pools (18 eggs per pool) were tested for the presence of 'R. felis-like' DNA. All samples were found positive by a specific ompAPCR assay, confirming the close association of this Rickettsia species with X. ramesis in all its life stages. These results suggest a symbiotic association between 'Rickettsia felis-like' and X. ramesis fleas.

Identification of different Bartonella species in the cattle tail louse (Haematopinus quadripertusus) and in cattle blood.

Bartonella spp. are worldwide-distributed facultative intracellular bacteria that exhibit an immense genomic diversity across mammal and arthropod hosts. The occurrence of cattle-associated Bartonella species was investigated in the cattle tail louse Haematopinus quadripertusus and in dairy cattle blood from Israel. Lice were collected from cattle from two dairy farms during summer 2011, and both lice and cow blood samples were collected from additional seven farms during the successive winter. The lice were identified morphologically and molecularly using 18S rRNA sequencing. Thereafter, they were screened for Bartonella DNA by conventional and real-time PCR assays using four partial genetic loci (gltA, rpoB, ssrA, and internal transcribed spacer [ITS]). A potentially novel Bartonella variant, closely related to other ruminant bartonellae, was identified in 11 of 13 louse pools collected in summer. In the cattle blood, the prevalence of Bartonella infection was 38%, identified as B. bovis and B. henselae (24 and 12%, respectively). A third genotype, closely related to Bartonella melophagi and Bartonella chomelii (based on the ssrA gene) and to B. bovis (based on the ITS sequence) was identified in a single cow. The relatively high prevalence of these Bartonella species in cattle and the occurrence of phylogenetically diverse Bartonella variants in both cattle and their lice suggest the potential role of this animal system in the generation of Bartonella species diversity.

Transmission dynamics of Bartonella sp. strain OE 1-1 in Sundevall's jirds (Meriones crassus).

A high prevalence of Bartonella infection is found in many natural systems; however, the transmission dynamics leading to observations of these infections is not fully understood. The capability of Xenopsylla ramesis fleas to serve as competent vectors of Bartonella sp. OE 1-1 (a strain closely related to the zoonotic Bartonella elizabethae) to Meriones crassus jirds was investigated. Naïve X. ramesis fleas were placed for 72 h on naïve jirds or jirds that were either experimentally or naturally infected with Bartonella sp. strain OE 1-1, after which they were placed on naïve jirds. Postfeeding, 69 to 100% of the fleas collected from each Bartonella-positive jird contained Bartonella DNA, and all naïve jirds became positive for Bartonella sp. OE 1-1 after infestation with the infected fleas. In addition, maternal transmission of Bartonella sp. OE 1-1 in jirds was tested by mating 5 Bartonella-positive and 5 naïve female jirds with 10 naïve male jirds in the absence of fleas. Fifteen offspring were delivered by each group. Cultures of blood drawn from all offspring on days 35 and 47 postdelivery were found to be negative for Bartonella. A single spleen sample from the offspring of a Bartonella-positive mother was found molecularly positive for Bartonella sp. OE 1-1. This study demonstrates that X. ramesis fleas are competent vectors of Bartonella sp. OE 1-1 to M. crassus jirds and indicates that maternal transmission is probably not the major transmission route from female jirds to their offspring. We suggest that the dynamics of Bartonella sp. OE 1-1 in the M. crassus jird population in nature is mostly dependent on its vectors.

Effects of Bartonella spp. on flea feeding and reproductive performance.

Numerous pathogens are transmitted from one host to another by hematophagous insect vectors. The interactions between a vector-borne organism and its vector vary in many ways, most of which are yet to be explored and identified. These interactions may play a role in the dynamics of the infection cycle. One way to evaluate these interactions is by studying the effects of the tested organism on the vector. In this study, we tested the effects of infection with Bartonella species on fitness-related variables of fleas by using Bartonella sp. strain OE 1-1, Xenopsylla ramesis fleas, and Meriones crassus jirds as a model system. Feeding parameters, including blood meal size and metabolic rate during digestion, as well as reproductive parameters, including fecundity, fertility, and life span, were compared between fleas experimentally infected with Bartonella and uninfected fleas. In addition, the developmental time, sex ratio, and body size of F1 offspring fleas were compared between the two groups. Most tested parameters did not differ between infected and uninfected fleas. However, F1 males produced by Bartonella-positive females were significantly smaller than F1 males produced by Bartonella-negative female fleas. The findings in this study suggest that bartonellae are well adapted to their flea vectors, and by minimally affecting their fitness they have evolved to better spread themselves in the natural environment.

Vertical nontransovarial transmission of Bartonella in fleas.

Pathogens use diverse pathways to infect host populations by vertical and/or horizontal routes. Horizontal transmission of bacteria belonging to the Bartonella genus via haematophagous vectors is well known. Vertical transmission of Bartonella species was also suggested to occur but its routes remain to be unveiled. In a previous study, we showed the absence of transovarial transmission of Bartonella species OE 1-1 in Xenopsylla ramesis fleas, and that fleas feeding on Bartonella-positive jirds produced Bartonella-positive gut voids. This current study aimed to investigate whether vertical nontransovarial transmission of Bartonella occurs in fleas. For this aim, the X. ramesis-Bartonella sp. OE 1-1 model was used. Four groups of fleas including Bartonella-positive and Bartonella-negative female fleas and larval offspring had access to either Bartonella-negative or Bartonella-positive gut voids and faeces. Sixteen per cent of flea offspring that had access to Bartonella-positive faeces and gut voids became Bartonella positive. Our findings demonstrate that Bartonella-positive flea faeces and gut voids are proper infection sources for flea larvae and indicate that vertical nontransovarial transmission of bartonellae occurs in fleas. This information broadens our understanding of Bartonella transmission routes in flea vectors and enlightens pathways of bartonellae transmission and maintenance in flea populations in nature.

Preliminary study of shark microbiota at a unique mix-species shark aggregation site, in the Eastern Mediterranean Sea.

Sharks, as apex predators, play an essential ecological role in shaping the marine food web and maintaining healthy and balanced marine ecosystems. Sharks are sensitive to environmental changes and anthropogenic pressure and demonstrate a clear and rapid response. This designates them a "keystone" or "sentinel" group that may describe the structure and function of the ecosystem. As a meta-organism, sharks offer selective niches (organs) for microorganisms that can provide benefits for their hosts. However, changes in the microbiota (due to physiological or environmental changes) can turn the symbiosis into a dysbiosis and may affect the physiology, immunity and ecology of the host. Although the importance of sharks within the ecosystem is well known, relatively few studies have focused on the microbiome aspect, especially with long-term sampling. Our study was conducted at a site of coastal development in Israel where a mixed-species shark aggregation (November-May) is observed. The aggregation includes two shark species, the dusky (Carcharhinus obscurus) and sandbar (Carcharhinus plumbeus) which segregate by sex (females and males, respectively). In order to characterize the bacterial profile and examine the physiological and ecological aspects, microbiome samples were collected from different organs (gills, skin, and cloaca) from both shark species over 3 years (sampling seasons: 2019, 2020, and 2021). The bacterial composition was significantly different between the shark individuals and the surrounding seawater and between the shark species. Additionally, differences were apparent between all the organs and the seawater, and between the skin and gills. The most dominant groups for both shark species were Flavobacteriaceae, Moraxellaceae, and Rhodobacteraceae. However, specific microbial biomarkers were also identified for each shark. An unexpected difference in the microbiome profile and diversity between the 2019-2020 and 2021 sampling seasons, revealed an increase in the potential pathogen Streptococcus. The fluctuations in the relative abundance of Streptococcus between the months of the third sampling season were also reflected in the seawater. Our study provides initial information on shark microbiome in the Eastern Mediterranean Sea. In addition, we demonstrated that these methods were also able to describe environmental episodes and the microbiome is a robust measure for long-term ecological research.

Identification of Mycobacterium pseudoshottsii in the Eastern Mediterranean.

Among the numerous pathogenic nontuberculous mycobacteria (NTM), which may cause disease in both poikilothermic and homoeothermic organisms, members of the unique clade Mycobacterium ulcerans/Mycobacterium marinum (MuMC) may cause disease in both fish and humans. Here, we describe the emergence of Mycobacterium pseudoshottsii, one of the four MuMC members, in Israel. For many years, M. marinum was the dominant NTM that was diagnosed in Israel as a fish pathogen. To the best of our knowledge, this is the first isolation and genomic characterization of M. pseudoshottsii infecting edible fish from two different fish species farmed in offshore sea cages in the eastern Mediterranean as well as in a recirculating aquaculture system in Israel. We compared the M. pseudoshottsii whole-genome sequences to all available genomic sequences of MuMC in free, publicly accessible databases. IMPORTANCE Mycobacterium pseudoshottsii was first detected in 1997 in the USA, infecting wild striped bass (Morone saxatilis). Since then, several reports from different countries worldwide have shown its capacity to become established in new regions as well as its pathogenicity to saltwater and euryhaline finfish of different genera. Our phylogenetic analysis revealed that the Mycobacterium ulcerans/Mycobacterium marinum clade (MuMC) is divided into two main branches: one that includes M. marinum and M. pseudoshottsii, and the second, which includes other M. marinum isolates as well as two isolates of M. shottsii. Our results reinforce the proposition that the geographical distribution of M. pseudoshottsii is much more extensive than is commonly believed. The emergence of M. pseudoshottsii in different parts of the world and its pathogenic traits that affect finfish of different genera may be a cause for concern among fish farmers, researchers, and environmental organizations.

First report on the serum chemistry and haematology of free-ranging dusky (Carcharhinus obscurus) and sandbar (Carcharhinus plumbeus) sharks in the eastern Mediterranean Sea.

Shark assessments in the Mediterranean Sea are still scarce, and serum chemistry and haematological data have yet to be reported for wild dusky (Carcharhinus obscurus) or sandbar (Carcharhinus plumbeus) shark populations in the Mediterranean Sea. Herein, blood samples were obtained from adult dusky (n = 23) and sandbar (n = 14) sharks from an aggregation site near the Hadera power and desalination plants in Israel in the winters of 2016-20. Several serum chemistry analytes were characterized with relation to stress, body size and environmental conditions. Glucose concentrations were higher, while total cholesterol concentrations were lower in dusky sharks than in sandbar sharks, potentially due to distinct metabolic pathways utilized during the capture-related activity by both species. However, differences in sex and size are noted and should be considered. The blood cell morphology of both species was consistent with previous findings for sandbar sharks. Atypical monocytes were noted in one dusky shark. Preliminary and exploratory reference intervals for female dusky sharks were calculated for glucose, triglycerides, total cholesterol, total protein and creatine kinase. These data must be viewed with caution due to the potential influence of capture-related stress on analyte concentrations and activities and the fact that only females were employed in the calculations. Moreover, the sampling site is adjacent to coastal power and desalination plants, which may significantly affect shark physiology. Although limited, this novel database on dusky and sandbar shark serum chemistry and haematology aspects is essential as a first attempt to obtain data on these species in the eastern Mediterranean Sea and for future conservation and long-term biomonitoring efforts.

Low prevalence of Contracaecum third-stage larvae parasitizing Sea of Galilee fisheries: 1-year survey after 57 years of no information.

Freshwater and marine ecosystems are a suitable habitat for parasitic nematodes of the genus Contracaecum (family: Anisakidae) to complete their complex life cycle. Several fish species of the Sea of Galilee (Lake Kinneret) were reported in 1964 as second intermediate/paratenic hosts of Contracaecum spp. larvae. The lack of taxonomically relevant morphological features of these larvae hindered their proper identification. Here we report the results of a 1-year survey conducted in 2021, 57 years after the first (and only) such survey. We analyzed 352 specimens from 10 fish species (native and non-native) of the Sea of Galilee (Israel) ichthyofauna. We compared our results with those of the first parasitological survey conducted by Paperna in 1964; the overall prevalence of nematodes referable to Contracaecum larvae was 16.8% and 0.85% in 1964 and in 2021, respectively. Different from the first survey that identified Contracaecum larvae morphologically, we used both morphological and molecular tools. Two wild native cyprinids-Jordan himri (Carasobarbus canis) and Jordan barbel (Luciobarbus longiceps)-were infected (a single specimen each) with Contracaecum quadripapillatum larvae in their abdominal cavity. A single specimen of blue tilapia (Oreochromis aureus) was infected with two larvae of Contracaecum multipapillatum E, localized in the pericardial cavity. The findings of our study, which is part of a large project focused on Contracaecum spp. infecting both piscivorous birds and fish collected in Israel, advance our knowledge about the distribution and host range of this potentially zoonotic parasite in fishery products of the Sea of Galilee.

Two Decades of Coastal Dolphin Population Surveys in Israel, Eastern Mediterranean.

Along the Mediterranean coast of Israel, two near-shore dolphin species are prevalent; Tursiops truncatus (least concern, IUCN) and Delphinus delphis (endangered, IUCN). Ship-board surveys and sporadic sightings over the last two decades have shown that the two differ in distribution-T. truncatus is found along the entire coast and D. delphis only in the south. The environmental and anthropological factors affecting these species' spatial distribution and determining their habitat preferences in this area are largely unknown. This work is a first attempt at summarizing 20 years of observations and studying habitat preferences for both species, by use of Generalized Additive Models. T. truncatus was found to be present in all areas of the continental shelf where survey effort coverage was sufficient, with a high affinity towards bottom trawlers. Model results showed D. delphis distribution to be associated to (shallow) water depths, though the factors driving their limited latitudinal distribution currently remain unknown. It is evident that T. truncatus and D. delphis are present in segregated areas of the Israeli continental shelf and T. truncatus currently sustains a delicate balance with continuously shifting human activities, while the drivers of D. delphis distribution are more specified, yet still not fully understood.

First description of a Gammaherpesvirus in a common dolphin (Delphinus delphis) from the Eastern Mediterranean Sea.

In September 2020, a male common dolphin (Delphinus delphis) was found dead on a beach near Bat-Yam, Israel. A small, raised, well circumscribed penile lesion (i.e., mass) was identified and removed for histology and molecular characterizations. By histology, the penile mass presented focal keratinization of the squamous epithelium and a mild ballooning of acanthocytes in lower epithelium levels, as well as features compatible with viral plaques, and tested positive for a gammaherpesvirus through molecular characterization analyses. Tissue samples from the lungs, liver, and spleen, however, tested negative for herpesvirus infection. The gammaherpesvirus detected herein is similar to other isolates found in several areas worldwide in different cetacean species. This is the first reported case of gammaherpesvirus infection in dolphins from the eastern Mediterranean Sea, indicative of the need for long-term assessments to create viral infections databases in cetaceans, especially in a climate change context, which is likely to intensify infectious disease outbreaks in marine mammals in the future.

Case report: Blindness associated with Learedius learedi trematode infection in a green sea turtle, Chelonia mydas, of the northern Red Sea.

Spirorchiid blood flukes are widespread in sea turtles, causing disease and mortality in their populations, with high prevalence in several ocean basins. Besides being leading parasitic causes of sea turtle strandings in several parts of the world, these infectious agents can cause endocarditis, vasculitis, thrombosis, miliary egg granulomas, and aneurysms, which ultimately may compromise the survival of green sea turtles. More severe cases may also result in multifocal granulomatous meningitis or pneumonia, both of which can be fatal. Herein, we report the first case of severe trematode infection, Caused by Learedius learedi, in a green sea turtle in the northern Red Sea; this infection is associated with bilateral blindness. Necropsy revealed multiple granulomas with intralesional trematode eggs in the optic nerve, eyes, spleen, heart, and lungs. The parasite was identified as Learedius learedi through specific primers of the ribosomal genome and COI sequences obtained from GenBank. Altogether, these findings emphasize the importance of recognizing the systemic nature of this particular fluke infection to ultimately protect the lives of these marine animals and ensure the sustainability of these species in the wild.

Unraveling Metabolite Provisioning to Offspring Through Parental Fluids: A Case Study of the Brazilian Guitarfish, Pseudobatos horkelii.

Elasmobranchs have a very distinct metabolism, and many aspects related to the energetic dynamics of these animals remain poorly investigated. The reproductive period is particularly vulnerable for viviparous species, as part of the energy reserves of the parental biomass is reallocated for gamete production and embryo development. In this context, this study aimed to characterize parental metabolite provisioning to the offspring (both sperm and developing embryos) of the Brazilian Guitarfish, Pseudobatos horkelii, through glucose, ß-hydroxybutyrate, triglycerides, and total cholesterol determinations in the uterine liquid (UL) and serum of pregnant females and in the seminal fluid (SF) and serum of males during the copulation period. No significant difference was observed for the analyzed markers between the UL and SF. Except for triglycerides, higher in female serum samples, all other energy markers were present at similar concentrations in the serum of both females and males. When comparing female UL and serum, significant differences were observed for triglycerides and total cholesterol. No differences were observed between SF and serum in males. The results indicate that all markers are being made available to offspring, possibly complementary to the yolk in the case of maternal liquid, and as an additional source for sperm mobilization required during egg fertilization in the case of the paternal fluid. Correlations between the markers in the parental matrices were also noted, compatible with the metabolic pathways activated during energy mobilization in vertebrates. Moreover, distinct marker predominance patterns were also noted for both UL and SF. Energy mobilization characterization directed to offspring through parental fluids aids in unraveling metabolic dynamics during the reproduction stage while also providing support for stress physiology studies to evaluate the indirect effects of parental allostatic overload in both sperm and developing embryos. Finally, energy mobilization assessments of parental fluids may also help elucidate how internal fertilization and viviparity evolved in this very distinct taxonomic group.

Morphological description and molecular characterization of Contracaecum larvae (Nematoda: Anisakidae) parasitizing market-size hybrid tilapia (Oreochromis aureus x Oreochromis niloticus) and red drum (Sciaenops ocellatus) farmed in Israel.

Nematodes belonging to the genus Contracaecum (family: Anisakidae) are heteroxenous parasites with a complex life cycle. Contracaecum larvae infecting farmed fish and fishery products are economically important causing market rejection in massive infection and may have zoonotic potential. In Israel, Contracaecum larvae have been described morphologically in several fish species; however, none of these descriptions were supported by molecular tools. In 2019-2020, hybrid tilapia (Oreochromis aureus x Oreochromis niloticus) and red drum (Sciaenops ocellatus), farmed in polyculture were found to be heavily infected with nematodes referable to Contracaecum larvae. Prevalence of infection in hybrid tilapia and red drum was 53.8% and 40.9%, respectively. A combined (morphological and molecular) approach revealed that both infected fish species were parasitized by the same species of Contracaecum, although larvae in hybrid tilapia were localized in the pericardial cavity whereas in red drum, they were observed in the abdominal cavity. Genetic analysis of internal transcribed spacer rDNA and cox2 mtDNA showed high similarity to unidentified Contracaecum larvae detected in several fish species in Ethiopia, Egypt and Kenya. In this study, molecular and morphological analyses place the possible new species in the C. multipapillatum complex and was provisionally named C. multipapillatum E. Further analyses combining morphological and molecular approaches are required on adult specimens collected from piscivorous birds living in the same area to support the identification of a potentially new species.