New insights into the gut microbiome in loggerhead sea turtles Caretta caretta stranded on the Mediterranean coast

Caretta caretta is the most common sea turtle species in the Mediterranean Sea. The species is threatened by anthropomorphic activity that causes thousands of deaths and hundreds of strandings along the Mediterranean coast. Stranded turtles are often cared for in rehabilitation centres until they recover or die. The objective of this study was to characterize the gut microbiome of nine sea turtles stranded along the Sicilian coast of the Mediterranean Sea using high-throughput sequencing analysis targeting V3–V4 regions of the bacterial 16S rRNA gene. Stool samples were collected from eight specimens hosted in the recovery centre after a few days of hospitalization (under 7) and from one ho…

The gas chromatography/mass spectrometry can be used for dose estimation in irradiated pork

Abstract Food safety can be improved using ionizing radiation to reduce food spoilage and to extend its shelf life. The gas chromatography/mass spectrometry (GC/MS) has been validated by the European Community as a powerful method to identify irradiated food containing fat. The preliminary goals of our research were: (i) to set up this method, based on the detection of radiation induced 2-dodecylcyclobutanones (2-DCB) in pork muscle samples and (ii) to check the microbiological efficacy of the treatment. The main objective was to render the GC/MS a quantitative technique for dose estimation, through the measurement of the 2-DCB concentration in the irradiated sample. Our results show that t…

Impact of Heavy Metals in Eggs and Tissues of C. caretta along the Sicilian Coast (Mediterranean Sea)

In this study we compared the heavy metal concentration found in different tissues and eggs of the loggerhead sea turtle and evaluated the potential ecotoxicological risk for this important species. Eighteen heavy metal elements were determined in different tissues (liver, gonads, fat, kidney, heart, brain, and spleen) of nine individuals of Caretta caretta found stranded along the coasts of Messina (Sicily, Italy) and in the shell and yolk of six eggs from the island of Linosa (Sicily, Italy). For the analysis of the heavy metals, we used the analytical procedures in accordance with the EPA 200.8 method supplemented by EPA 6020b with three replicates for each measurement. The elements anal…

A retrospective study of the characterization of Rickettsia species in ticks collected from humans

Rickettsiae (family Rickettsiaceae, order Rickettsiales) are obligate intracellular bacteria transmitted by arthropod vectors. Several Rickettsia species causing vector-borne rickettsioses belong to the spotted fever group (SFG). Traditionally, Rickettsia conorii has been considered as the main etiologic agent of Mediterranean spotted fever. However, the molecular characterization of rickettsiae allowed identifying other species involved in spotted fever in the Mediterranean region. In this study, 42 ticks collected from humans were subjected to morphological identification and molecular characterization of Rickettsia species potentially involved in human rickettsiosis in Sicily. Fourteen t…

Can DNA methylation contribute to explain morphological variability? The case of loggerhead turtle Caretta caretta

The loggerhead turtle Caretta caretta is a common and threatened sea turtle in the Mediterranean Sea. It is listed in the Red List of Threatened Species of the International Union for Conservation of Nature (IUCN 2012) and, although it is widely studied, little is currently known about the relationships between its phenotype and genetic variability. In the last few years some authors observed the presence of individuals, collected in different parts of the Mediterranean Sea, with a variable number of carapacial and plastron scutes. This variability, in some cases, makes difficult the sound identification of the species (Margaritoulis and Chiras, 2011; Turkozan et al. 2001; Oliver 2014). To …

New records of leatherback sea turtle, Dermochelys coriacea (Vandelli, 1761) (Testudines: Dermochelyidae) in the Strait of Sicily

We report three new records of leatherback sea turtle, Dermochelys coriacea (Vandelli, 1761) recovered in February 2011 and in spring-summer 2016 in the Strait of Sicily. The individuals were handed over dead to the IZS of Palermo for necropsy. The first specimen measured 139 and 91 cm of CCL and CCW respectively and was found beached and alive but died a few hours later for septicemia. The second measured 110 cm of CCL and 83 cm of CCW and showed a deep wound on its head probably due to contact with a boat propeller. The third individual measured 134 cm of CCL and 96 cm of CCW and had plastic bags, a piece of fishing net and debris of various nature in the stomach. For the second individua…

Pedigree verification with multiple candidate parents using Pedverif software

Species like sheep and beef cattle are commonly raised in large herds and often on pasture with multiple sires joining the females for unrecorded natural insemination. This leads to offspring with multiple candidate parents and therefore uncertain parentage. Twins or triplets can be from multiple sires as well. Pedigrees from such populations are often problematic and need proper verification. The Pedverif computer program is able to verify normal pedigrees as well as pedigrees with multiple candidate parents.

A new record and molecular characterization of the Green Turtle Chelonia mydas (Testudines, Cheloniidae) in Sicilian coastal waters

A juvenile specimen of the Green Turtle, Chelonia mydas, was recovered in Sicilian coastal waters close to Avola (Syracuse province, south-eastern Sicily, Italy). Before being released, the specimen was measured and inspected for the presence of ingested hooks or other possible harms to its health. A fragment of the mtDNA marker COI was amplified and sequenced in order to provide the first molecular data for the species from the central Mediterranean area. The possible influence of climate change on the occurrence of this thermophilic species in the central Mediterranean area is briefly discussed.

Antigen-specific T cells and cytokines detection as useful tool for understanding immunity against zoonotic infections.

Zoonoses include a broad range of diseases, that are becoming of great interest, due to the climate changing, that cause the adaptation of vectors to new niches and environments. Host immune responses play a crucial role in determining the outcome of infections, as documented by expansion of antigen-specific T cells during several zoonotic infections. Thus, understanding of the contribution of antigen-specific T-cell subsets in the host immune response is a powerful tool to evaluate the different immunological mechanisms involved in zoonotic infections and for the development of effective vaccines. In this paper we discuss the role of T cells in some eukaryotic and prokaryotic infectious mo…

Incidental catch of loggerhead sea turtles (Caretta caretta) along the Sicilian coasts by longline fishery

It has been estimated that 44,000 Caretta caretta turtles die every year due to anthropomorphic activity in the Mediterranean Sea, and that longline fishing is one of the most significant causes of mortality. A total of 482 specimens of C. caretta were rescued from different parts of the Sicilian coast (Mediterranean Sea) from 2014 to 2016. The most numerous stranding was recorded during the spring and summer seasons, mainly along the north and eastern coasts of Sicily. The curved carapace length for all the specimens ranged from between 19 and 95 cm and most of these were young or sub adults. The highest number of strandings was recorded in 2014 and 2015, with 206 and 169 individuals, resp…

Analysis of spatial diffusion of ferric ions in PVA-GTA gel dosimeters through magnetic resonance imaging

Abstract This work focused on the analysis of the temporal diffusion of ferric ions through PVA-GTA gel dosimeters. PVA-GTA gel samples, partly exposed with 6 MV X-rays in order to create an initial steep gradient, were mapped using magnetic resonance imaging on a 7T MRI scanner for small animals. Multiple images of the gels were acquired over several hours after irradiation and were analyzed to quantitatively extract the signal profile. The spatial resolution achieved is 200 μm and this makes this technique particularly suitable for the analysis of steep gradients of ferric ion concentration. The results obtained with PVA-GTA gels were compared with those achieved with agarose gels, which …

Prion protein gene frequencies in three Sicilian dairy sheep populations

The objective of this paper was to investigate the prion protein (PrP) genotype and haplotype frequencies in three Sicilian dairy sheep populations. The three populations were: (1) 1096 Valle del Belice animals, (2) 1143 Comisana animals, and (3) 1771 individuals from 5 flocks with scrapie outbreaks, in which the animals were crossbreds derived from indigenous Sicilian dairy breeds. PrP genotypes are described for the three codons 136 (Alanine or Valine; A, V), 154 (Histidine or Arginine; H, R), and 171 (Glutamine, Arginine or Histidine; Q, R, H) which represent polymorphisms known to be linked with scrapie susceptibility. The Valle del Belice haplotype frequencies were 32.3% ARR, 6.5% AHQ,…

Toxocara canis infection induces antigen-specific IL-10 and IFNγ production in pregnant dogs and their puppies

Toxocara canis (T. canis) is originally a parasite of canine bitches and their pups. The pathogenicity of T. canis infection is enhanced during pregnancy and puppyhood. The aim of this study was to investigate if modification of IFNgamma and IL-10 secretion occurs during infection in pregnant dogs and puppies. Analysis of cytokines secreted could let us hypothesize a role for IL-10 and/or IFNgamma in T. canis infection. We tested T. canis-specific production of IFNgamma and IL-10 by lymphocytes of pregnant dogs and their puppies after in vitro re-exposure to purified excretory/secretory antigen (ESAg) from T. canis. Blood mononuclear cells (BMC) isolated from pregnant dogs and their puppies…

ESAT-6 Peptide Recognition by Bovine CD8 + Lymphocytes of Naturally Infected Cows in Herds from Southern Italy

ABSTRACT The aim of this study was to define epitopes of Mycobacterium bovis from ESAT-6 (early secretory antigen of 6 kDa) recognized by CD8 + T lymphocytes from cows naturally infected with Mycobacterium bovis . We found that bovine CD8 + T cells recognized 10 out of 11 ESAT-6 peptides tested.

Characterization of the Apical Membrane Antigen-1 in Italian Strains of Babesia bigemina

Summary Babesia bigemina is a parasite endemic in different parts of the world, including Europe and the Americas. One of the few genes characterized in this species codifies for the Apical Membrane Antigen 1 (AMA-1), a trans-membrane antigen recently identified. In this research, we characterized the ama-1 gene from three Italian B. bigemina strains, two B. bigemina strains obtained from Ragusa, Sicily (ITA1 and ITA3) and a third one obtained from Benevento, Campania (ITA2). Italian sequences were compared with those of the Australian strain obtained from the Sanger Institute web site and to strains from different parts of the world. The results obtained confirmed that this newly described…

Nonmodal scutes patterns in the Loggerhead Sea Turtle (Caretta caretta): a possible epigenetic effect?

Eleven specimens of the threatened Loggerhead Sea Turtle (Caretta caretta (L., 1758)) were caught accidentally by fishermen in different parts of Sicily (Mediterranean Sea). Five of them showed an atypical number of carapacial and plastron scutes, making the immediate identification of the specimens as C. caretta difficult. Both genetic and epigenetic analysis were carried out on these specimens. Sequencing of a 649 bp sequence of the mitochondrial cytochrome c oxidase I (COI) gene allowed us to classify all the individuals as C. caretta. Epigenetic analysis, performed by evaluating the total level of DNA cytosine methylation, showed a reduced and significant (F = 72.65, p < 0.01) globa…

Molecular characterization of dominant bacterial population in “Vastedda della Valle del Belice” cheese: preliminary investigation

The sensory characteristics of raw-milk cheeses are linked to the cheese-making process, to the environmental factors as animal feeding and to the biochemical and microbiological composition of the milk. In this report we temped to characterize the microflora in the typical Sicilian historical cheese as Vastedda della valle del Belice. Each cheese was previous subjected to microbial isolation on specific media (M17 and MRS). The colony obtained on the solid medium were subject to biochemical tests and DNA extraction. The microbial diversity occurring in the strains was evaluated by PCR, RFLP and sequencing targeted on 16S ribosomal DNA. A number of closest relatives species of lactic and co…

Nutraceutical effects of table green olives: a pilot study with Nocellara del Belice olives

Background: The aim of this study was to analyse the nutraceutical properties of table green olives Nocellara del Belice, a traditional Mediterranean food. The Mediterranean Diet has as key elements olives and extra virgin olive oil, common to all Mediterranean countries. Olive oil is the main source of fat and can modulate oxidative stress and inflammation, whereas little is known about the role of olives. Moreover, emerging evidences underline the association between gut microbiota and food as the basis of many phenomena that affect health and delay or avoid the onset of some age-related chronic diseases. Methods: In order to show if table green olives have nutraceutical properties and/or…

Stranding patterns of the striped dolphin (Stenella coeruleoalba , Meyen 1833, Delphinidae) along the Sicilian coast (Mediterranean Sea)

Development and validation of two PCR tests for the detection of and differentiation between Anaplasma ovis and Anaplasma marginale

Anaplasma ovis and Anaplasma marginale are tick-transmitted bacteria that cause anaplasmosis in domestic and wild animals. Recent results show that some domestic and wild animals and ticks are susceptible to both A. ovis and A. marginale, thus supporting the need to differentiate between these species in hosts and ticks diagnosed with Anaplasma infection. However, although anaplasmosis is one of the most common diseases of grazing animals worldwide, rapid and effective tests are not available for the detection of and discrimination between these 2 Anaplasma species. The objective of this research was to develop an easy and reliable method to identify and discriminate between the closely rel…

Detection of natural killer T cells in mice infected with Rickettsia conorii.

Little information is available regarding the role of natural killer T (NKT) cells during the early stage of Rickettsia conorii infection. Herein, C3H/HeN mice were infected with the Malish 7 strain of R. conorii. Splenocytes from these mice were analysed in the early stage of the infection by flow cytometry and compared with uninfected controls. Our results showed an increase in NKT cells in infected mice. Additionally, NKT interleukin (IL)-17(+) cells increased three days after infection, together with a concurrent decrease in the relative amount of NKT interferon (IFN)-γ(+) cells. We also confirmed a higher amount of NK IFN-γ(+) cells in infected mice. Taken together, our data showed tha…

First data on microflora of loggerhead sea turtle (Caretta caretta) nests from the coastlines of Sicily (Italy)

ABSTRACT Caretta caretta is threatened by many dangers in the Mediterranean basin, but most are human-related. The purposes of this research were: (i) to investigate microflora in samples from six loggerhead sea turtle nests located on the Sicilian coast and (ii) to understand microbial diversity associated with nests, with particular attention to bacteria and fungi involved in failed hatchings. During the 2016 and 2018 summers, 456 eggs and seven dead hatchling from six nests were collected. We performed bacteriological and mycological analyses on 88 egg samples and seven dead hatchlings, allowing us to isolate: Fusarium spp. (80.6%), Aeromonas hydrophila (55.6%), Aspergillus spp. (27.2%) …

Codominance of Lactobacillus plantarum and obligate heterofermentative lactic acid bacteria during sourdough fermentation

Fifteen sourdoughs produced in western Sicily (southern Italy) were analysed by classical methods for their chemico-physical characteristics and the levels of lactic acid bacteria (LAB). pH and total titratable acidity (TTA) were mostly in the range commonly reported for similar products produced in Italy, but the fermentation quotient (FQ) of the majority of samples was above 4.0, due to the low concentration of acetic acid estimated by high performance liquid chromatography (HPLC). Specific counts of LAB showed levels higher than 10(8) CFU g(-1) for many samples. The colonies representing various morphologies were isolated and, after the differentiation based on phenotypic characteristics…

Chasing phthalates in tissues of marine turtles from the Mediterranean sea

Tissues from thirteen specimens of marine turtles, one Dermochelys coriacea and twelve Caretta caretta, found dead along the Sicilian coasts in 2016 were analyzed for the presence of phthalates. Four phthalates (DEP, DBP, BBP, and DEHP) were found at different significant concentrations in liver and gonads, while only DBP was found in muscle tissues and at a fourfold lower concentration than other phthalates in Dermochelys coriacea. No traces of DEP were detected in C. caretta tissues where DOTP was also revealed. The presence of phthalates in fat tissue in specimens of C. caretta showed a major prevalence of the most lipophilic phthalates DEHP and DOTP. The total concentration of all analy…

Nuevo registro y caracterización molecular de la tortuga verde Chelonia mydas (Testudines, Cheloniidae) en las aguas costeras sicilianas

A juvenile specimen of the Green Turtle, Chelonia mydas, was recovered in Sicilian coastal waters close to Avola (Syracuse province, south-eastern Sicily, Italy). Before being released, the specimen was measured and inspected for the presence of ingested hooks or other possible harms to its health. A fragment of the mtDNA marker COI was amplified and sequenced in order to provide the first molecular data for the species from the central Mediterranean area. The possible influence of climate change on the occurrence of this thermophilic species in the central Mediterranean area is briefly discussed.