The First Report of Pennella (Crustacea: Copepoda) Infesting Stenella coeruleoalba Stranded in Malta: Morphological and Genetic Analyses.

Here, we document the stranding of a striped dolphin Stenella coeruleoalba (Meyen, 1833) (Mammalia: Delphinidae), which was found dead in Maltese waters in July 2020. The stranded dolphin exhibited a severe infestation of the mesoparasitic copepod, Pennella balaenoptera Koren and Danielssen, 1877 (Copepoda: Pennelidae). Parasites of this genus represent the largest known mesoparasites to infest cetaceans. Under normal circumstances, cetaceans may have a few P. balaenoptera individuals attached to them, but cetaceans with compromised health are more prone to heavy infestations. The identification of the parasite was accomplished through morphological and genetic analyses. This incident highlights the significance of monitoring mesoparasitic infestations, offering valuable insights into the health of cetacean populations and emphasizing the potential implications for conservation efforts in the region.

A pan-cetacean MHC amplicon sequencing panel developed and evaluated in combination with genome assemblies.

The major histocompatibility complex (MHC) is a highly polymorphic gene family that is crucial in immunity, and its diversity can be effectively used as a fitness marker for populations. Despite this, MHC remains poorly characterised in non-model species (e.g., cetaceans: whales, dolphins and porpoises) as high gene copy number variation, especially in the fast-evolving class I region, makes analyses of genomic sequences difficult. To date, only small sections of class I and IIa genes have been used to assess functional diversity in cetacean populations. Here, we undertook a systematic characterisation of the MHC class I and IIa regions in available cetacean genomes. We extracted full-length gene sequences to design pan-cetacean primers that amplified the complete exon 2 from MHC class I and IIa genes in one combined sequencing panel. We validated this panel in 19 cetacean species and described 354 alleles for both classes. Furthermore, we identified likely assembly artefacts for many MHC class I assemblies based on the presence of class I genes in the amplicon data compared to missing genes from genomes. Finally, we investigated MHC diversity using the panel in 25 humpback and 30 southern right whales, including four paternity trios for humpback whales. This revealed copy-number variable class I haplotypes in humpback whales, which is likely a common phenomenon across cetaceans. These MHC alleles will form the basis for a cetacean branch of the Immuno-Polymorphism Database (IPD-MHC), a curated resource intended to aid in the systematic compilation of MHC alleles across several species, to support conservation initiatives.

Probiotics in Infancy and Childhood for Food Allergy Prevention and Treatment.

Food allergy represents a failure of oral tolerance mechanisms to dietary antigens. Over the past few years, food allergies have become a growing public health problem worldwide. Gut microbiota is believed to have a significant impact on oral tolerance to food antigens and in initiation and maintenance of food allergies. Therefore, probiotics have also been proposed in this field as a possible strategy for modulating both the gut microbiota and the immune system. In recent years, results from preclinical and clinical studies suggest a promising role for probiotics in food allergy prevention and treatment. However, future studies are needed to better understand the mechanisms of action of probiotics in food allergies and to design comparable study protocols using specific probiotic strains, defined doses and exposure times, and longer follow-up periods.

A species-level trait dataset of bats in Europe and beyond.

Knowledge of species' functional traits is essential for understanding biodiversity patterns, predicting the impacts of global environmental changes, and assessing the efficiency of conservation measures. Bats are major components of mammalian diversity and occupy a variety of ecological niches and geographic distributions. However, an extensive compilation of their functional traits and ecological attributes is still missing. Here we present EuroBaTrait 1.0, the most comprehensive and up-to-date trait dataset covering 47 European bat species. The dataset includes data on 118 traits including genetic composition, physiology, morphology, acoustic signature, climatic associations, foraging habitat, roost type, diet, spatial behaviour, life history, pathogens, phenology, and distribution. We compiled the bat trait data obtained from three main sources: (i) a systematic literature and dataset search, (ii) unpublished data from European bat experts, and (iii) observations from large-scale monitoring programs. EuroBaTrait is designed to provide an important data source for comparative and trait-based analyses at the species or community level. The dataset also exposes knowledge gaps in species, geographic and trait coverage, highlighting priorities for future data collection.

Conservation Genetics of the Loggerhead Sea Turtle, Caretta caretta, from the Central Mediterranean: An Insight into the Species' Reproductive Behaviour in Maltese Waters.

Loggerhead sea turtle, Caretta caretta (Linnaeus, 1758), nestlings were investigated through specimens found dead either after hatching or unhatched (n = 120) from eight nests around the Maltese islands (Central Mediterranean). Molecular genetics was used to conduct maternity and paternity tests of the collected specimens utilizing expanded mitochondrial DNA sequences from the control region (858 bp) and 25 microsatellite loci (12 dinucleotide loci and 13 tetranucleotide loci). Mitochondrial data produced two haplotypes, CC-A2.1 and CC-A3.1, with the most common haplotype being present in seven nests. Microsatellite data revealed the identity of six different females that were involved in the deposition of the eggs in the eight turtle nests analysed. This confirms that two females laid multiple nests. Additionally, microsatellite data allowed for the determination of multiple paternity, with one clutch being sired by two fathers. These results are useful for monitoring the genetic diversity of loggerhead sea turtle nestlings and of the turtle mothers and fathers contributing to future turtle offspring, which rely on Maltese sandy beaches for their successful start to life. Effective conservation management benefits from merging scientific knowledge with effective measures at potential nesting sites to avoid losses of nestlings caused by human negligence.

Role of Vitamin D in Celiac Disease and Inflammatory Bowel Diseases.

Vitamin D (VD) is a pro-hormone that has long been known as a key regulator of calcium homeostasis and bone health in both children and adults. In recent years, studies have shown that VD may exert many extra-skeletal functions, mainly through a relevant modulation of the innate and adaptive immune system. This has suggested that VD could play a fundamental role in conditioning development, clinical course, and treatment of several autoimmune disorders, including celiac disease (CD) and inflammatory bowel diseases (IBDs). The main aim of this review is to evaluate the relationships between VD, CD, and IBDs. Literature analysis showed a potential impact of VD on CD and IBDs can be reasonably assumed based on the well-documented in vitro and in vivo VD activities on the gastrointestinal tract and the immune system. The evidence that VD can preserve intestinal mucosa from chemical and immunological damage and that VD modulation of the immune system functions can contrast the mechanisms that lead to the intestinal modifications characteristic of gastrointestinal autoimmune diseases has suggested that VD could play a role in controlling both the development and the course of CD and IBDs. Administration of VD in already diagnosed CD and IBD cases has not always significantly modified disease course. However, despite these relevant problems, most of the experts recommend monitoring of VD levels in patients with CD and IBDs and administration of supplements in patients with hypovitaminosis.

Management of Breast Abscess during Breastfeeding.

(1) Background: Breast abscess (BA) is a condition leading in the majority of cases to breastfeeding interruption. Abscesses are commonly treated with antibiotics, needle aspiration or incision and drainage (I&D), but there is still no consensus on the optimal treatment. Since there are no well-defined clinical guidelines for abscess management, we conducted a retrospective, observational study with the aim of assessing ultrasound (US)-guided management of BA without surgery, regardless of the BA size. The secondary objective was the microbiologic characterization and, in particular, the S. aureus methicillin resistance identification. (2) Methods: our population included 64 breastfeeding mothers with diagnosis of BA. For every patient, data about maternal, perinatal and breastfeeding features were collected. All patients underwent office US scans and 40 out of 64 required a more detailed breast diagnostic ultrasound performed by a radiologist. In all cases, samples of milk or abscess material were microbiologically tested. All patients received oral antibiotic treatment. We performed needle aspiration, when feasible, even on abscesses greater than 5 cm. (3) Results: most of the women developed BA during the first 100 days (68.8% during the first 60 days) after delivery and 13 needed hospitalization. Four abscesses were bilateral and 16 had a US major diameter greater than 5 cm. All patients were treated with antibiotic therapy according to our clinical protocol and 71.9% (46/64) underwent fine needle aspiration. None of them required I&D. The average duration of breastfeeding was 5 months (IR 2; 9.5) and 40.6% of women with BA continued to breastfeed for more than 6 months. Only 21 mothers interrupted breastfeeding before 3 months. (4) Conclusions: our observational data suggest, regardless of the size and the clinical features of the BA, a conservative approach with antibiotic therapy targeted at the Methicillin-Resistant Staphilococcus aureus (MRSA) identified and needle aspiration, if feasible. In our experience, treatment with needle aspiration is a cost- effective method. Unlike drainage, it is an outpatient procedure, easily repeatable, with no cosmetic damage. In addition, it has lower risk of recurrences since, differently from surgical incision, it does not cause interruption of the ducts. Moreover, needle aspiration is less painful, does not require the separation of the mother-child dyad and allows for a quicker, if not immediate, return to breastfeeding.

The first complete mitochondrial genomes for Serranus papilionaceus and Serranus scriba, and their phylogenetic position within Serranidae.

BACKGROUND: Butterfly-winged comber, Serranus papilionaceus Valenciennes, 1832, was recently resurrected and so it is no longer considered as a junior synonym of the Painted comber, Serranus scriba (Linneus, 1758). This calls for a more comprehensive phylogenetic assessment using mitochondria DNA genomes to better understand the relationship and delineate these two species. METHODS AND RESULTS: Next-generation Sequencing was applied to sequence the genome of these two Serranus species. The data generated was then used to construct the mitochondrial genome of these two species. This produced the first complete mitochondrial genomes for the genus Serranus here represented by Serranus papilionaceus and Serranus scriba. These two mitochondrial genomes are 16,514 bp and 16,512 bp respectively, and both contained the typical 37 genes found in vertebrates (13 protein-coding genes, two ribosomal RNAs, 22 transfer RNAs), together with the OL and the control region. CONCLUSIONS: These mitochondrial genomes provide a new insight into the phylogenetic and evolutionary connections between the various subfamilies within Serranidae, while providing new molecular data that can be applied to discriminate between the studied species.

The importance of the patient's perspective in function-sparing parotid surgery for benign neoplasms: clinical reappraisal.

OBJECTIVE: Function-sparing surgery is the cornerstone for the treatment of benign parotid neoplasms. We assessed the incidences and determinants of the main postoperative complications, reappraising their influence on the patient's quality of life (QoL). METHODS: Patients who underwent parotid surgery for benign neoplasms were reviewed (2016-2019). Parotidectomy Outcome Inventory-8 (POI-8) and condition-specific questionnaires were used to investigate the patient's perspective. RESULTS: We enrolled 211 patients. Preservation of the posterior branch of the great auricular nerve (GAN) seemed to reduce early dysfunction (87% vs 96%, p = 0.053), but not the late one. Deep lobe dissection and resection of more than one parotid segment favoured first bite syndrome (FBS) and Frey's syndrome (FS), respectively (16% vs 3%, p = 0.003; 37% vs 15%, p = 0.003). Neither GAN impairment, FBS, nor FS influenced patient QoL. Facial weakness affected 19 patients (9%), being more likely after total parotidectomy (23% vs 7%, p = 0.034). According to POI-8, QoL was mainly jeopardised by fear of revision surgery, especially in females (p = 0.005) and those experiencing early complications (p = 0.004). CONCLUSIONS: Reappraisal of the patient's perspective after functional parotid surgery is fundamental to tailor preoperative counselling.

Characterization of the complete mitogenome of Haifa grouper, Hyporthodus haifensis (Perciformes: Serranidae), and its phylogenetic position within Epinephelini.

The complete mitochondrial genome of the Haifa grouper, Hyporthodus haifensis (Ben-Tuvia, 1953), has been obtained, through Illumina next-generation sequencing, and annotated. This mitogenome was found to be 16,525 bp long and to contain 37 genes, a control region, and the L-strand replication origin. The overall base composition of the complete mitogenome for this species was found to be 28.55% A, 28.07% C, 16.32% G, and 27.06% T. This study also looked into the mitogenome phylogenetic relationships of H. haifensis within the tribe Epinephelini and adds to the genetic resources currently available for the species.

Resurrection of the Butterfly-winged Comber, Serranus papilionaceus Valenciennes, 1832 (Teleostei, Serranidae) and its phylogenetic position within genus Serranus.

The family Serranidae is represented by 92 genera and 579 valid species, with the genus Serranus Cuvier, 1816, containing 30 species. In this study, specimens of Butterfly-winged Comber, Serranus papilionaceus Valenciennes, 1832, were collected from the Canary Islands and compared morphologically and genetically to Painted Comber, Serranus scriba (Linnaeus, 1758), from the Mediterranean Sea. Morphological differences, especially in the colour banding pattern, were corroborated by genetic differences in mitochondrial (COI and ND2) and nuclear (Rhod and PTR) markers. The mitochondrial DNA markers revealed a high level of divergence and no shared haplotypes between the two species (interspecific divergence: COI 4.31%; ND2 8.68%), and a phylogenetic analysis showed that these two species are closely related sister species sharing common ancestry. This study is therefore offering to resurrect S. papilionaceus Valenciennes, 1832 as a valid species increasing the number of eastern Atlantic Serranus species to 11. This should direct new species-specific research, including its population conservation status assessment across its distribution.

Characterization and comparison of the complete mitochondrial genomes of two stingrays, Dasyatis pastinaca and Dasyatis tortonesei (Myliobatiformes: Dasyatidae) from the Mediterranean Sea.

This work represents the complete mitochondrial genomes of two stingrays, Dasyatis pastinaca and Dasyatis tortonesei, from the Mediterranean Sea. The mitogenomes of these two species were obtained through whole generation sequencing and annotated. These mitogenomes were found to be 17,713 bp and 17,630 bp respectively and each contained 37 genes, that is 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes and two ribosomal RNA (rRNA) genes, a control region and the origin of L-strand replication (OL). All PCGs begin with the codon ATG, except for cytochrome c oxidase subunit 1 (COX1) gene, while two PCGs end with an incomplete termination codon. All tRNA genes are able to fold into their typical cloverleaf secondary structures, except for tRNA-Ser1AGY which lacks the dihydrouracil arm. The control region was 1982 bp and 1889 bp for D. pastinaca and D. tortonesei respectively. Phylogenetic analysis using Bayesian Inference confirmed the occurrence of the genus Dasyatis within the monophyletic subfamily Dasyatinae. This study adds on genetic resources available for these two stingray species.

The complete mitogenome of the Critically Endangered smalltooth sand tiger shark, Odontaspis ferox (Lamniformes: Odontaspididae).

Here, we report the first complete mitochondrial genome for the smalltooth sand tiger shark, Odontaspis ferox (Risso, 1810). The circular mitochondrial genome was found to be 16,682 bp in length and contains 37 genes, a control region and the replication origin of the L-strand (OL). The base composition of this mitogenome is 32.6% A, 23.3% C, 12.8% G, and 31.3% T. Phylogenetic analysis of Lamniformes indicates that O. ferox did not group with Carcharias taurus and so the taxonomic classification of Odontaspididae needs to be revised. This study promotes conservation genetics for this poorly studied shark species which is listed critically endangered in the Mediterranean Sea.

Assessing cetacean surveys throughout the Mediterranean Sea: a gap analysis in environmental space.

Heterogeneous data collection in the marine environment has led to large gaps in our knowledge of marine species distributions. To fill these gaps, models calibrated on existing data may be used to predict species distributions in unsampled areas, given that available data are sufficiently representative. Our objective was to evaluate the feasibility of mapping cetacean densities across the entire Mediterranean Sea using models calibrated on available survey data and various environmental covariates. We aggregated 302,481 km of line transect survey effort conducted in the Mediterranean Sea within the past 20 years by many organisations. Survey coverage was highly heterogeneous geographically and seasonally: large data gaps were present in the eastern and southern Mediterranean and in non-summer months. We mapped the extent of interpolation versus extrapolation and the proportion of data nearby in environmental space when models calibrated on existing survey data were used for prediction across the entire Mediterranean Sea. Using model predictions to map cetacean densities in the eastern and southern Mediterranean, characterised by warmer, less productive waters, and more intense eddy activity, would lead to potentially unreliable extrapolations. We stress the need for systematic surveys of cetaceans in these environmentally unique Mediterranean waters, particularly in non-summer months.

A molecular approach towards taxonomic identification of elasmobranch species from Maltese fisheries landings.

The mitochondrial genome, through the application of DNA barcoding, provides a powerful tool for identifying species even when specimens are either incomplete or belong to species that exhibit cryptic diversity. In fisheries management accurate identification of whole or part of the specimens landed is a fundamental requirement for the conservation of species affected directly or indirectly by the fisheries activities. In this study cytochrome c oxidase subunit I (COI) and NADH dehydrogenase subunit 2 (ND2) sequences were used to genetically distinguish 36 elasmobranch species collected from Maltese (Central Mediterranean) commercial fisheries landings. Each species was analysed using these two mtDNA loci where COI (610bp) and ND2 (990bp) efficiently distinguished between the various species studied, leading to the identification of 101 haplotypes, with the intraspecific p-distance ranging between 0 and 0.75% (mean 0.10%, SD ±0.13%). This study enhances the molecular data available on elasmobranchs by providing new ND2 sequences for various species, while providing both COI and ND2 data for poorly studied Mediterranean species including: the large pelagic sharks Alopias vulpinus, A. superciliosus, Carcharhinus altimus, C. plumbeus, Carcharadon carcharias, Isurus oxyrinchus, Prionace glauca and Odontaspis ferox; the smaller demersal sharks Somniosus rostratus, Squatina aculeata, S. oculata and Squalus sp.; and the endemic stingray Dasyatis tortonesei. It also confirmed the landings of species whose identification relies strongly on molecular tools, namely Squalus sp. and D. tortonesei, which are both first confirmed records amongst Maltese fisheries landings. Morphologically, the latter two species, can be easily misidentified with S. blainville and D. pastinaca respectively. Additionally, this study evaluated the genetic differences between different polychromatic forms of Raja clavata, R. radula and Dipturus oxyrinchus. Based on the currently analysed specimens, no significant genetic differences were found between the various forms and thus no further speciation within the species was identified.

Non-perforated peptic ulcer disease: multidetector CT findings, complications, and differential diagnosis.

Despite availability of effective therapies, peptic ulcer disease (PUD) remains a major global disease, resulting from a combination of persistent Helicobacter pylori infection and widespread use of nonsteroidal anti-inflammatory drugs. Albeit endoscopy definitely represents the mainstay diagnostic technique, patients presenting to emergency departments with unexplained abdominal pain generally undergo multidetector CT as an initial investigation. Although superficial ulcers generally remain inconspicuous, careful multiplanar CT interpretation may allow to detect deep ulcers, secondary mural and extraluminal signs of peptic gastroduodenitis, thereby allowing timely endoscopic verification and appropriate treatment. This pictorial essay aims to provide radiologists with an increased familiarity with CT diagnosis of non-perforated PUD, with emphasis on differential diagnosis. Following an overview of current disease epidemiology and complications, it explains the appropriate CT acquisition and interpretation techniques, and reviews with several examples the cross-sectional findings of uncomplicated PUD. Afterwards, the CT features of PUD complications such as ulcer haemorrhage, gastric outlet obstruction, biliary and pancreatic fistulisation are presented. TEACHING POINTS: • Gastric and duodenal peptic ulcers are increasingly caused by nonsteroidal anti-inflammatory drugs • Multiplanar CT interpretation allows detecting deep ulcers and secondary signs of gastroduodenitis • CT diagnosis of uncomplicated peptic disease relies on direct and indirect signs • Currently the commonest complication, haemorrhage may be treated with transarterial embolisation • Other uncommon complications include gastric outlet obstruction and biliopancreatic fistulisation.

Population genetics of the deep-sea bluntnose sixgill shark, Hexanchus griseus, revealing spatial genetic heterogeneity.

Hexanchus griseus is a globally distributed deep-water shark species. It inhabits tropical and temperate waters throughout the world, including the Mediterranean Sea where it is by-caught by small-scale fisheries in the region. In this study, we analysed the genetic variation of H. griseus specimens collected from different areas within and outside the Mediterranean region, to assess its genetic connectivity. The mitochondrial DNA (mtDNA) sequence analysed in this study ranged from cytochrome b to 16S rRNA genes including the control region, the 12S rRNA gene and the interspersed tRNA genes in the region, covering a total of 3731 to 3914 nucleotides. Results have shown that this species exhibits geographically distinct maternal lineages, indicating population structure along geographical ranges. These findings reveal population subdivisions not only between the Pacific Ocean and the Atlantic Ocean, but also within the oceans and on a smaller scale within the Mediterranean Sea. This highlights the need to consider each population subdivision separately when designing management plans for the conservation of this species.

Population Structure, Genetic Diversity, Effective Population Size, Demographic History and Regional Connectivity Patterns of the Endangered Dusky Grouper, Epinephelus marginatus (Teleostei: Serranidae), within Malta's Fisheries Management Zone.

The objective of this study is to describe the genetic population structure and demographic history of the endangered marine fish, Epinephelus marginatus, within Malta's Fisheries Management Zone for the purpose of localised conservation planning. Epinephelus marginatus is a long-lived, sedentary, reef-associated protogynous hermaphrodite with high commercial and recreational value that is at risk of extinction throughout its global distribution. Based on global trends, population substructuring and gaps in local knowledge this has led to an increased interest in evaluation of local stock. Assessment of Maltese demography was based on historical and contemporary catch landings data whilst genetic population structure and regional connectivity patterns were evaluated by examining 175 individuals collected within the central Mediterranean region between 2002 and 2009 using 14 nuclear microsatellite loci. Demographic stock assessment of Maltese E. marginatus' revealed a 99% decline in catch landings between 1947 and 2009 within the Fisheries Management Zone. A contemporary modest mean size was observed, 3 ± 3 kg, where approximately 17% of the population was juvenile, 68% female/sex-changing and 15% were male with a male-to-female sex ratio of 1:5. Genetic analysis describes the overall population of E. marginatus' within the Fisheries Management Zone as decreasing in size (ƟH = 2.2), which has gone through a significant size reduction in the past (M = 0.41) and consequently shows signs of moderate inbreeding (FIS = 0.10, p < 0.001) with an estimated effective population size of 130 individuals. Results of spatially explicit Bayesian genetic cluster analysis detected two geographically distinct subpopulations within Malta's Fisheries Management Zone and that they are connected to a larger network of E. marginatus' within the Sicily Channel. Results suggest conservation management should be designed to reflect E. marginatus' within Malta's Fisheries Management Zone as two management units.