A ‘machine learning’ technique for discriminating captive-reared from wild Atlantic bluefin tuna, Thunnus thynnus (Osteichthyes: Scombridae), based on differential fin spine bone resorption

The Atlantic bluefin tuna (ABFT) fishery is regulated by the International Commission for the Conservation of Atlantic Tunas (ICCAT), which establishes the allowable annual yield and the minimum capture size, and allocates capture quotas to the Contracting Parties. Despite fishery monitoring, a considerable amount of captures escapes ICCAT control. In the Mediterranean Sea, the purse seine fishery supports ABFT farming, a capture-based aquaculture activity that involves catching fish from the wild and rearing them in sea cages for a few months. The first spine of the cranial dorsal fin undergoes a continuous bone remodeling process consisting in old bone (primary bone) resorption and new bone (secondary bone) apposition. A marked increase of spine bone resorption was shown in captive-reared ABFT with respect to wild specimens. In this paper, the Random Forest (RF), a Computer Aided Detection system, was applied to distinguish captive-reared from wild ABFT based on fish age, fish fork length, total surface of spine cross section, and surface of remodeled bone tissue in the spine cross section (sum of reabsorbed bone tissue and secondary cancellous bone). The RF system was also compared to the Logistic Regression method (LR). The percentages of properly classified animals, either wild or captive-reared, with respect to the overall number of animals, i.e. accuracy, was 95.3 ± 2.6% and 79.0 ± 5.1% for RF and LR, respectively. The percentages of the properly classified captive-reared specimens, i.e. sensitivity, were 93.5 ± 3.1% and 75.8 ± 5.3% for RF and LR, respectively. The percentages of the properly classified wild specimens was 96.7 ± 2.2% and 81.4 ± 4.9%, for RF and LR, respectively. The proposed technique appears to be a reliable investigation tool anytime the suspicion arises that illegally caught ABFT are sold as aquaculture products

Modulation of Morphology and Glycan Composition of Mucins in Farmed Guinea Fowl (Numida meleagris) Intestine by the Multi-Strain Probiotic Slab51®

Probiotics have become highly recognized as supplements for poultry.Since gut health can be considered synonymous withanimal health, the effects of probiotic Slab51® on the morphology and the glycan composition of guineafowlintestine were examined. The probiotics were added in drinking water (2 x 1011 UFC/L) throughout the grow-out cycle.Birds were individually weighed andslaughtered after four months. Samples from the duodenum, ileum and caecum were collected and processed for morphological, morphometric, conventional and lectin glycohistochemical studies. The results were analyzed for statistical significance by Student’s t test. Compared with control samples, probiotic group revealed (1) significant increase in villus height (p < 0.001 in duodenum and ileum; p < 0.05 in caecum), crypt depth (p < 0.001 in duodenum and caecum; p < 0.05 in ileum) and goblet cells (GCs) per villus (p < 0.001) in all investigated tracts; (2) increase in galactosel,3Nacetylgalacyosamine( Gall,3GalNAc)terminating O-glycans and l,2-fucosylated glycans secretory GCs in the duodenum; (3) increase in 2,6-sialoglycans and high-mannose N-linked glycans secretory GCs but reduction in GCs-secreting sulfoglycans in the ileum; (4) increase in Gall,3GalNAc and high-mannose N-linked glycans secretory GCs and decrease in GCs-producing sulfomucins in the caecum; (5) increase in the numbers of crypt cells containing sulfate and non-sulfated acidic glycans. Overall, dietary Slab51® induces morphological and region-specific changes in glycoprotein composition of guinea fowl intestine, promoting gut health

Positive Influence of a Probiotic Mixture on the Intestinal Morphology and Microbiota of Farmed Guinea Fowls (Numida meleagris)

To understand the effectiveness of a probiotic mixture on intestinal morphology, mucus layer composition, and cecal microbiota diversity, 40 10-day-old Guinea fowls (Numida meleagris) were assigned to two groups: the control group (C), receiving drinking water, and the treated group (P), receiving water plus a commercial multi-strain probiotic (Slab51®, 2 × 1011 CFU/L). Birds were slaughtered after 4 months, and the intestines were collected. Samples from the duodenum, ileum, and cecum were processed for morphological and morphometric studies, and conventional glycohistochemistry. Cecal samples were also used to assess the microbiota by 16S metataxonomic approach. Group P showed significant increase in the villus height (p < 0.001 in the duodenum and p < 0.05 in the ileum and cecum), villus width (p < 0.05 in all investigated tracts), depth of crypts (p < 0.001 in the duodenum and cecum; p < 0.05 in the ileum), and goblet cells per villus (p < 0.001 in all investigated tracts) compared with group C. Cecal microbiota of the birds varied considerably and comparing the relative abundance of the main observational taxonomic units (OTUs), a positive enrichment of several beneficial taxa, such as Oscillospira, Eubacterium, Prevotella, and members of the Ruminococcaceae, was observed. The enrichment of those taxa can improve microbiota stability and resilience facing environmental stresses, enhancing its resistance against invading pathogens. Ruminococcaceae, which represent the most important taxon in both groups, and Prevotella have a key role in the gut physiology due to the production of short-chain fatty acids (SCFAs), which are a vital energy source for enterocytes, improve glucose metabolism, and exert an overall anti-inflammatory effect. Probiotic administration enriches the presence of Coprococcus, Oscillospira, and Eubacterium taxa that produce butyrate, which exerts a beneficial effect on growth performance, structure of villi, and pathogen control and has anti-inflammatory properties too. This study indicates that Slab51® supplementation positively affects the morphology and microbiota diversity of the guinea fowl intestine

Recent Insights on the Maternal Microbiota: Impact on Pregnancy Outcomes

Abstract Hormonal changes during and after pregnancy are linked with modifications in the maternal microbiota. We describe the importance of the maternal microbiota in pregnancy and examine whether changes in maternal microbiotic composition at different body sites (gut, vagina, endometrium) are associated with pregnancy complications. We analyze the likely interactions between microbiota and the immune system. During pregnancy, the gastrointestinal (gut) microbiota undergoes profound changes that lead to an increase in lactic acid-producing bacteria and a reduction in butyrate-producing bacteria. The meaning of such changes needs clarification. Additionally, several studies have indicated a possible involvement of the maternal gut microbiota in autoimmune and lifelong diseases. The human vagina has its own microbiota, and changes in vaginal microbiota are related to several pregnancy-related complications. Recent studies show reduced lactobacilli, increased bacterial diversity, and low vaginal levels of beta-defensin 2 in women with preterm births. In contrast, early and healthy pregnancies are characterized by low diversity and low numbers of bacterial communities dominated by Lactobacillus. These observations suggest that early vaginal cultures that show an absence of Lactobacillus and polymicrobial vaginal colonization are risk factors for preterm birth. The endometrium is not a sterile site. Resident endometrial microbiota has only been defined recently. However, questions remain regarding the main components of the endometrial microbiota and their impact on the reproductive tract concerning both fertility and pregnancy outcomes. A classification based on endometrial bacterial patterns could help develop a microbiota-based diagnosis as well as personalized therapies for the prevention of obstetric complications and personalized treatments through nutritional, microbiotic, or pharmaceutical interventions. Keywords: endometrium; gut; immunity; inflammasome; microbiota; pregnancy; vagina

Fin Spine Bone Resorption in Atlantic Bluefin Tuna, Thunnus thynnus, and Comparison between Wild and Captive-Reared Specimens

Bone resorption in the first spine of the first dorsal fin of Atlantic bluefin tuna (ABFT) has long been considered for age estimation studies. In the present paper spine bone resorpion was assessed in wild (aged 1 to 13 years) and captive-reared (aged 2 to 11 years) ABFT sampled from the Mediterranean Sea. Total surface (TS), solid surface (SS) and reabsorbed surface (RS) were measured in spine transverse sections in order to obtain proportions of SS and RS. The spine section surface was found to be isometrically correlated to the fish fork length by a power equation. The fraction of solid spine bone progressively decreased according to a logarithmic equation correlating SS/TS to both fish size and age. The values ranged from 57% in the smallest examined individuals to 37% in the largest specimens. This phenomenon was further enhanced in captive-reared ABFT where SS/TS was 22% in the largest measured specimen. The difference between the fraction of SS of wild and captive-reared ABFT was highly significant. In each year class from 1- to 7-year-old wild specimens, the fraction of spine reabsorbed surface was significantly higher in specimens collected from March to May than in those sampled during the rest of the year. In 4-year-old fish the normal SS increase during the summer did not occur, possibly coinciding with their first sexual maturity. According to the correlations between SS/TS and age, the rate of spine bone resorption was significantly higher, even almost double, in captive-reared specimens. This could be attributed to the wider context of systemic dysfunctions occurring in reared ABFT, and may be related to a number of factors, including nutritional deficiencies, alteration of endocrine profile, cortisol-induced stress, and loss of spine functions during locomotion in rearing conditions.Versión del editor4,411

Gaia Early Data Release 3: Structure and properties of the Magellanic Clouds

We compare the Gaia DR2 and Gaia EDR3 performances in the study of the Magellanic Clouds and show the clear improvements in precision and accuracy in the new release. We also show that the systematics still present in the data make the determination of the 3D geometry of the LMC a difficult endeavour; this is at the very limit of the usefulness of the Gaia EDR3 astrometry, but it may become feasible with the use of additional external data. We derive radial and tangential velocity maps and global profiles for the LMC for the several subsamples we defined. To our knowledge, this is the first time that the two planar components of the ordered and random motions are derived for multiple stellar evolutionary phases in a galactic disc outside the Milky Way, showing the differences between younger and older phases. We also analyse the spatial structure and motions in the central region, the bar, and the disc, providing new insights into features and kinematics. Finally, we show that the Gaia EDR3 data allows clearly resolving the Magellanic Bridge, and we trace the density and velocity flow of the stars from the SMC towards the LMC not only globally, but also separately for young and evolved populations. This allows us to confirm an evolved population in the Bridge that is slightly shift from the younger population. Additionally, we were able to study the outskirts of both Magellanic Clouds, in which we detected some well-known features and indications of new ones

Multiple-Phase Biometric Relationships and Sexual Maturity in the Atlantic Bluefin Tuna, <i>Thunnus thynnus</i> (Osteichthyes: Scombridae)

Most fish undergo distinct growth phases during ontogenesis. An extremely important passage from the juvenile to adult phase occurs at the onset of sexual maturity, which shows in body proportion and/or growth rate changes. These can be detected as change-points in biometric relationships. In this paper, the Atlantic bluefin tuna was analyzed to verify whether its somatic proportions show any sign of discontinuity during growth, i.e., whether any change-points may be detected in its somatic proportions. This fish has never been examined in this respect, and single-phase models, which are indeed easier to both compute and apply, are used in stock analyses. The following somatic relationships were analyzed in Atlantic bluefin tuna captured in the Mediterranean Sea between 1998 and 2010: “fork length–weight” regression, the von Bertalanffy growth equation, and “first dorsal spine cross section surface–fork length” regression. All of the examined relationships were found to be best modelled by multiple-phase regression equations, and all of them showed a change-point within the range of 101–110 cm fork length, which corresponds to 3–4 years of age. The present results, based on reproductive state-independent analyses, corroborate the disputed hypothesis that Atlantic bluefin tuna from the eastern stock in fact reproduce for the first time at this age

In situ characterization of glycans in the horse bladder urothelium

The urinary bladder cavity is lined by a highly specialized epithelium, the urothelium, which prevents permeation of solutes and noxious agents back into the bloodstream and underlying tissues, serving also as a sensor and transducer of physiological and nociceptive stimuli [1]. In addition, in physiological conditions the urothelium can also function as a secretory tissue [2]. A mucous layer protects the urothelium of the urinary bladder from potentially harmful environmental substances, attachment of bacteria and proteolytic enzymes present in the urine [3]. Moreover, the glycan composition of urothelial mucous layer could have a role in the intravesical pharmacological treatments [4]. Despite their considerable importance in urinary bladder physiology, few studies are available about the glycoconjugates expressed in non-human species. In this study the glycoconjugate pattern of urothelium lining the horse urinary bladder was investigated. Tissue fragments from three horse stallions in good health status, aged 2.5-4 years, were fixed in 4% (w/v) PBS-buffered paraformaldehyde, embedded in paraffin wax and stained with a panel of twelve lectins, in combination with saponification and sialidase digestion (Ks). The urinary bladder urothelium has three distinct layers from the basal zone to the lumen consisting of basal, intermediate and superficial cells (umbrella cells). Cytoplasm of basal cells showed glycans ending with Neu5AcGal1-3GalNAc, GlcNAc and with terminal/internal Man (Ks-PNA, GSA II, and Con A II reactivity). A sub-population of intermediate cells also displayed terminal Neu5Ac2-6Gal/GalNAc, NeuNacα2-3Galβ1- 4GlcNAc, Gal1-3GalNAc, Gal, terminal and sialic acid-linked GalNAc, internal GlcNAc and Fucα1-2Galβ1-4GlcNAc (MAL II, SNA, PNA, GSA I-B4, SBA, ks-SBA, Ks-WGA, and UEA I reactivity). The cytoplasm of umbrella cell population contained all the above cited sugar residues. Moreover, LTA-reactive fucosylated glycans and Ks-DBA-positive sialoderivatives were found in some scattered umbrella cells. These sialoglycans were secreted in the bladder lumen. The bladder luminal surface stained with MAL II, SNA, PNA, Ks-PNA, and GSA I-B4 displaying a coating of sialo- and galactose-terminating glycoconjugates. These findings show that different glycosylation patterns exist along the horse bladder urothelium, and different sub-populations of umbrella cells are present secreting the sialoglycans which constitute the protective gel layer lining the bladder. Compared to results of a similar study carried out onthe donkey bladder urothelium [2], the present research reveals a species-specific glycan pattern and could contribute to a better understanding of the differences between domesticated odd-toed ungulate mammals via comparative glycopattern investigation. [1] Lasič et al. Properties of the urothelium that establish the blood-urine barrier and their implications for drug delivery, Reviews of Physiology, Biochemistry and Pharmacology Vol 168, 2015. [2] Desantis et al. In situ characterization of glycans in the urothelium of donkey bladder: Evidence of secretion of sialomucins, Acta Histochemica, 115:712–718, 2013. [4] Lopedota et al. Spray dried chitosan microparticles for intravesical delivery of celecoxib: preparation and characterization, Pharmaceutical Research, 33:2195–2208, 2016

Atti SISVET 2021

The urinary bladder cavity is lined by a highly specialized epithelium, the urothelium, which prevents permeation of solutes and noxious agents back into the bloodstream and underlying tissues, serving also as a sensor and transducer of physiological and nociceptive stimuli[1]. In addition, in physiological conditions the urothelium can also function as a secretory tissue[2]. A mucous layer protects the urothelium of the urinary bladder from potentially harmful environmental substances, attachment of bacteria and proteolytic enzymes present in the urine[3]. Moreover, the glycan composition of urothelial mucous layer could have a role in the intravesical pharmacological treatments[4]. Despite their considerable importance in urinary bladder physiology, few studies are available about the glycoconjugates expressed in non-human species. In this study the glycoconjugate pattern of urothelium lining the horse urinary bladder was investigated. Tissue fragments from three horse stallions in good health status, aged 2.5-4 years, were fixed in 4% (w/v) PBS-buffered paraformaldehyde, embedded in paraffin wax and stained with a panel of twelve lectins, in combination with saponification and sialidase digestion (Ks). The urinary bladder urothelium has three distinct layers from the basal zone to the lumen consisting of basal, intermediate and superficial cells (umbrella cells). Cytoplasm of basal cells showed glycans ending with Neu5AcGal1-3GalNAc, GlcNAc and with terminal/internal Man (Ks-PNA, GSA II, and Con A II reactivity). A sub-population of intermediate cells also displayed terminal Neu5Ac2-6Gal/GalNAc, NeuNacα2-3Galβ1-4GlcNAc, Gal1-3GalNAc, Gal, terminal and sialic acid-linked GalNAc, internal GlcNAc and Fucα1-2Galβ1-4GlcNAc (MAL II, SNA, PNA, GSA I-B4, SBA, ks-SBA, Ks-WGA, and UEA I reactivity). The cytoplasm of umbrella cell population contained all the above cited sugar residues. Moreover, LTA-reactive fucosylated glycans and Ks-DBA-positive sialoderivatives were found indifferent scattered umbrella cells. These sialoglycans were secreted in the bladder lumen. The bladder luminal surface stained with MAL II, SNA, PNA, Ks-PNA, and GSA I-B4 displaying a coating of sialo- and galactose-terminating glycoconjugates. These findings show that different glycosylation patterns exist along the horse bladder urothelium, and different sub-populations of umbrella cells are present secreting the sialoglycans which constitute the protective gel layer lining the bladder. Compared to results of a similar study carried out onthe donkey bladder urothelium[2], the present research reveals a species-specific glycan pattern and could contribute to a better understanding of the differences between domesticated odd-toed ungulate mammals via comparative glycopattern investigation. [1] Lasič et al. Properties of the urothelium that establish the blood-urine barrier and their implications for drug delivery, Reviews of Physiology, Biochemistry and Pharmacology Vol 168, 2015. [2] Desantis et al. In situ characterization of glycans in the urothelium of donkey bladder: Evidence of secretion of sialomucins, Acta Histochemica, 115:712–718, 2013. [3] Kreft et al. Formation and maintenance of blood–urine barrier in urothelium, Protoplasma, 246:3–14, 2010. [4] Lopedota et al. Spray dried chitosan microparticles for intravesical delivery of celecoxib: preparation and characterization, Pharmaceutical Research, 33:2195–2208, 2016