Does Anisakis spp. infestation affect the proximate composition, fatty acids, and minerals contents of its host Merluccius merlucccius?

Fish play an important role in human nutrition. They are not only a great source of protein and healthy fats, but also a unique source of essential nutrients such as omega-3 fatty acids. Moreover, most fish are parasitized, and some of these parasites are able to influence the reallocation of resources in their favor and thus reduce the nutritional quality of the fish. The present study was conducted to investigate the impact of the third stage larvae (L3) of Anisakis spp. on the proximate composition, macro-minerals (potassium, calcium, and sodium), and fatty acids of European hake (Merluccius merluccius Linnaeus, 1758). In parasitized female group, our results revealed a decrease (p< 0.005) in the amount of carbohydrate by 6.5%, of calcium by 17%, and of 2 polyunsaturated fatty acids (arachidonic acid (C20: 4w-6), and eicosapentaenoic acid (C20: 5w-3) with 33% and 15% respectively. Simultaneously, an increase by 25% in the level of a single saturated fatty acid C10:0 was noticed. According to the principal component analysis, the parasitized female was wealthy of saturated fatty acids and monounsaturated fatty acids and contains less of polyunsaturated fatty acids, omega-3 fatty acids, and omega-6 fatty acids than the unparasitized female and male. No significant changes were observed in the biochemical composition of male hake, probably due to the low mean intensity of L3 larvae of Anisakis spp. in this group.

Morphological and molecular studies of the life-cycle stages of the monorchiid Monorchis parvus (Looss, 1902) (Digenea) from the Southern Mediterranean coast (Tunisia).

The elucidation of life-cycles of digeneans, with their successive larval stages, is facilitated by the use of molecular markers. Samples of sporocysts containing cercariae and metacercariae belonging to Monorchis Monticelli, 1893 were collected from naturally infected bivalves, Cerastoderma glaucum (Bruguière, 1789), and adult forms of Monorchis spp. were collected from sparid fishes of the genus Diplodus. All specimens were collected in the Gulf of Gabès, southern Tunisia. The identities of the examined molluscs and fishes were determined via molecular barcoding of their COI gene. Sequences of COI and ITS1 genes were also obtained for both larval and adult stages of collected parasite specimens. Genetic sequence data generated for the collected larval specimens only differed minimally from the sequence data of adults identified as Monorchis parvus; we attribute the difference to intraspecific variation. The morpho-anatomical study showed that the different stages of M. parvus collected from the Tunisian coasts had the same morphology as those reported in European waters with a lag in maturity and lower measurements. The species is recorded and molecularly characterised for the first time off the Tunisian coasts.

Echinocephalus caniculus n. sp. (Nematoda: Gnathostomatidae Railliet, 1895) from the lesser spotted dogfish Scyliorhinus canicula (L.) (Elasmobranchii: Scyliorhinidae Gill, 1862) off Tunisia, with a key to species of the genus Echinocephalus.

Echinocephalus caniculus n. sp. (Nematoda, Gnathostomatidae Railliet, 1895) was isolated from the spiral valve of the lesser spotted dogfish Scyliorhinus canicula (L.) from the waters off Kalaat El Andalous, North East Tunisia. This new species is mainly characterized by a cephalic bulb armed with 31-39 transverse rows of uncinated hooks, a comparatively long oesophagus, short spicules and the presence of a gubernaculum. The new species differs from its congeners by having four cervical sacs of almost equal length, a higher oesophagus/body length ratio, the arrangement of the caudal papillae, the absence of a medioventral preanal organ and numerous scattered `pores´ limited to the lateral side of the posterior part of the body. This is the first report of a member of the genus Echinocephalus Molin, 1858 from the Tunisian coast, and a new host and locality record for the Gnathostomatidae. A key to the species of Echinocephalus is provided.

Genetic Approaches for Definitive Diagnosis of Agammaglobulinemia in Consanguineous Families.

Autosomal recessive agammaglobulinemia (ARA) is a primary immunodeficiency characterized by absent peripheral B cells, severe hypogammaglobulinemia, and absent BTK gene mutations. In ARA, mutations occur in genes encoding the pre-B cell receptor (pre-BCR) or downstream signaling proteins. In this work, we used candidate gene and whole-exome sequencing to investigate the molecular basis of ARA in 6 patients from 4 consanguineous North-African families. Sanger sequencing of candidate genes encoding the pre-BCR components (ΙGΗΜ, CD79A, CD79B, IGLL1, and VPREB1) was initially performed and determined the genetic defect in five patients. Two novel mutations in IGHM (p.Val378Alafs*1 and p.Ile184Serfs*21) were identified in three patients from two unrelated kindred and a novel nonsense mutation was identified in CD79A (p.Trp66*) in two siblings from a third kindred. Whole-exome sequencing (WES) was performed on the sixth patient who harbored a homozygous stop mutation at position 407 in the RAG2 gene (p.Glu407*). We concluded that conventional gene sequencing, especially when multiple genes are involved in the defect as is the case in ARA, is costly and time-consuming, resulting in delayed diagnosis that contributes to increased morbidity and mortality. In addition, it fails to identify the involvement of novel and unsuspected gene defects when the phenotype of the patients is atypical. WES has the potential to provide a rapid and more accurate genetic diagnosis in ARA, which is crucial for the treatment of the patients.

First description of the male and subgravid female of Philometra serranellicabrillae Janiszewska, 1949 (Nematoda: Philometridae), a gonad-infecting parasite of the comber Serranus cabrilla (Linnaeus) (Serranidae) off Tunisia.

The male and subgravid female of Philometra serranellicabrillae Janiszewska, 1949 (Philometridae) collected from the gonads of Serranus cabrilla (Linnaeus) (Serranidae) off Tunisia are described for the first time based on light and scanning electron microscopical studies. The male of this nematode can be separated from other congeneric gonad-infecting nominal species in the structure and shape of the gubernaculum (e.g. absence of a dorsal protuberance and a median smooth field). The shape of the male posterior region is unique in that it bears a pair of big circular papillae posterior to the cloacal opening, which is also present in other Philometra spp. from serranids, i.e. P. indica Moravec & Manoharan, 2014, P. inexpectata Moravec, Chaabane, Justine & Neifar, 2016 and P. jordanoi (López-Neyra, 1951) Yamaguti, 1961. Moreover, P. serranellicabrillae differs from its congeners in other fish families from the Mediterranean Sea, in the length of spicules and gubernaculum.

Two new gonad-infecting species of Philometra Costa, 1845 (Nematoda: Philometridae) from Trachinus spp. (Osteichthyes: Trachinidae) in the Gulf of Hammamet, Tunisia.

Based on light and scanning electron microscopical studies, two new gonad-infecting species of Philometra Costa, 1845, P. draco n. sp. and P. radiata n. sp. (Nematoda: Philometridae), are described from the marine perciform fishes Trachinus draco (Linnaeus) and T. radiatus (Linnaeus) (both Trachinidae), respectively, in the Gulf of Hammamet, off the northeastern coast of Tunisia. Philometra draco n. sp. and P. radiata n. sp. can be separated from other gonad-infecting species of this genus by the structures associated to the gubernaculum (e.g. dorsal protuberance, smooth field separating the dorsolateral longitudinal parts), as well as by the length of the body, spicules and gubernaculum. Philometra radiata n. sp. can be distinguished from P. draco n. sp. in having the dorsal side of the gubernaculum distal end provided with a median longitudinal smooth field demarcated by two dorsolateral lamellate parts. These two new species are the first philometrid species described from fishes of the family Trachinidae.

Morphological variability in Acanthocephaloides irregularis Amin, Oguz, Heckmann, Tepe & Kvach, 2011 (Acanthocephala: Arhythmacanthidae) from the European sea bass, Dicentrarchus labrax (L.) (Moronidae) in Bizerte Lagoon, Tunisia.

Specimens of an arhythmacanthid acanthocephalan were recovered from the European sea bass Dicentrarchus labrax (L.) (Perciformes: Moronidae) in Bizerte Lagoon, northern Tunisia. The specimens collected showed high morphological similarities to Acanthocephaloides irregularis Amin, Oguz, Heckmann, Tepe & Kvach, 2011, recovered from four species of marine fishes in the Gulf of Odessa and Sukhyi Lyman off the Ukrainian Black Sea coast. Light and scanning electron microscopy revealed some morphological differences from the Black Sea specimens in trunk spine distribution, number of proboscis hooks in males (4 instead of 5), and lack of demonstrable trunk collar. These variations suggest that our specimens may only be a morphotype of A. irregularis with new host and locality records. A comparison between our specimens and those of the original description clarifies the possible reasons of this intraspecific morphological variability.

Co segregation of the m.1555A>G mutation in the MT-RNR1 gene and mutations in MT-ATP6 gene in a family with dilated mitochondrial cardiomyopathy and hearing loss: A whole mitochondrial genome screening.

Mitochondrial disease refers to a heterogeneous group of disorders resulting in defective cellular energy production due to dysfunction of the mitochondrial respiratory chain, which is responsible for the generation of most cellular energy. Because cardiac muscles are one of the high energy demanding tissues, mitochondrial cardiomyopathies is one of the most frequent mitochondria disorders. Mitochondrial cardiomyopathy has been associated with several point mutations of mtDNA in both genes encoded mitochondrial proteins and mitochondrial tRNA and rRNA. We reported here the first description of mutations in MT-ATP6 gene in two patients with clinical features of dilated mitochondrial cardiomyopathy. The mutational analysis of the whole mitochondrial DNA revealed the presence of m.1555A>G mutation in MT-RNR1 gene associated to the m.8527A>G (p.M>V) and the m.8392C>T (p.136P>S) variations in the mitochondrial MT-ATP6 gene in patient1 and his family members with variable phenotype including hearing impairment. The second patient with isolated mitochondrial cardiomyopathy presented the m.8605C>T (p.27P>S) mutation in the MT-ATP6 gene. The three mutations p.M1V, p.P27S and p.P136S detected in MT-ATP6 affected well conserved residues of the mitochondrial protein ATPase 6. In addition, the substitution of proline residue at position 27 and 136 effect hydrophobicity and structure flexibility conformation of the protein.

Redescription of Philometra filiformis (Stossich, 1896) (Nematoda: Philometridae), a gonad-infecting parasite of the common pandora Pagellus erythrinus (Linnaeus) (Sparidae) in the Mediterranean Sea, including new taxonomic features revealed by SEM.

The insufficiently known nematode species Philometra filiformis (Stossich, 1896) (Philometridae) is redescribed based on light and scanning electron microscopical (SEM) examinations of specimens collected from the ovaries of the type-host, the common pandora Pagellus erythrinus (Linnaeus) (Perciformes, Sparidae), from off the Mediterranean coast of Tunisia. The male of this species was studied with SEM for the first time, which revealed some new, taxonomically important morphological features. The male posterior end has a caudal mound consisting of two lateral parts widely separated dorsally from each other, four pairs of small adanal papillae, a pair of large but moderately developed papillae located posterior to the cloacal aperture and a pair of small phasmids. The distal end of the gubernaculum is unique among all but one gonad-infecting species of Philometra Costa, 1845 by the chevron-shaped dorsal lamellar structures forming a median longitudinal range. Philometra filiformis is for the first time reported from Tunisian coastal waters, being the seventh philometrid species so far recorded from marine fishes off Tunisia.

Neonatal lupus erythematosus with congenital heart block in twins.

Background - Neonatal lupus erythematosus is an uncommon acquired autoimmune disease caused by transplacental passage of maternal antibodies SSA/Ro, SSB/La or U1 ribonucleoproteins. The most common clinical manifestations are skin rash, cardiac lesions, thrombocytopenia, anemia and hepatosplenomegaly. Complete congenital heart block is usually irreversible needing a pacemaker implantation in two-thirds of cases. Cases report - We report neonatal lupus erythematosus with complete congenital heart block in twins. Newborns were delivered by caesarean section at week 38 of gestation with a heart rate regular at 70 beats per minute. Both twins and mother were positive for antinuclear, anti-SSA, and anti-SSB antibodies. Twins received single-chamber pacemaker implants at day 12 of life. The evolution was immediately favorable with a heart rate around 110 beats per minute. The follow-up was 2 years. The twins are currently asymptomatic. Conclusion - Complete congenital heart block is the most serious manifestation of the neonatal lupus erythematosus associated with significant morbidity and mortality.

Molecular evidence for the existence of species complexes within Macvicaria Gibson & Bray, 1982 (Digenea: Opecoelidae) in the western Mediterranean, with descriptions of two new species.

Morphological and molecular characterisation of specimens of Macvicaria spp. (Digenea: Opecoelidae) from five species of Mediterranean sparids (Teleostei: Sparidae) sampled in the Bizerte Lagoon and the Bay of Bizerte (Tunisia) revealed the presence of two species complexes designated as "obovata" and "crassigula" groups. Detailed comparative morphological and phylogenetic analyses revealed that two of the specimen sets, one from each complex, represent species new to science. M. maamouriae n. sp. from Sparus aurata L. and Lithognathus mormyrus (L.) appeared genetically similar to M. obovata (Molin, 1859) but differs in having a much larger ventral sucker relative to body width, a cirrus-sac extending dorsally to the posterior margin of the ventral sucker or more posterior, vitelline fields comprising distinctly more abundant vitelline follicles, reaching the level of the pharynx both ventrally and dorsally and confluent dorsally in the forebody, and an ovary contiguous with the anterior testis. Macvicaria bartolii n. sp. from Diplodus annularis (L.) and Spondyliosoma cantharus (L.) belongs to the "crassigula" group and is characterised by having almost exclusively dorsal vitelline follicles forming two non-confluent fields in the forebody. Additional morphological data are provided for Macvicaria dubia (Stossich, 1905), a poorly known parasite of Oblada melanura (L.). Morphological descriptions are associated with ITS1-5.8S-ITS2 and 28S rDNA sequences for the three Mediterranean species of Macvicaria Gibson & Bray, 1982. The results of our study suggest that further exploration of the species diversity within this genus should be based on both morphological and molecular data.

Congenital heart disease in 37,294 births in Tunisia: birth prevalence and mortality rate.

AIM: To investigate the previously unknown birth incidence, treatment, and mortality of children with congenital heart disease in Tunisia. METHODS: We undertook a retrospective review of medical records of all patients who were born in 2010 and 2011, and were diagnosed in Sfax (Tunisia) with congenital heart defect. RESULTS: Among 37,294 births, 255 children were detected to have congenital heart disease, yielding a birth incidence of 6.8 per 1000. The most frequently occurring conditions were ventricular septal defects (31%), ostium secundum atrial septal defects (12.9%), and pulmonary valve abnormalities (12%). Coarctation of the aorta, tetralogy of Fallot, univentricular physiology, pulmonary atresia with ventricular septal defect, and transposition of the great arteries were found in 4.3%, 6.2%, 3.4%, 2.7%, and 2.7%, respectively. During the follow-up of 1 year, 23% of the children died. About three-quarters of those deaths happened before surgery. CONCLUSION: The present study is in line with the general estimates in the world. It has revealed a high case of mortality among the patients awaiting corrective surgery. These children need more facilities.

Trace elements assessment in Cerastoderma glaucum from port areas in the Tunisian Mediterranean coast: The influence of parasites on bioaccumulation.

In the present study, the seasonal concentration of seven trace elements was investigated in sediment and the cockle Cerastoderma glaucum tissues from two port areas in the North-East and South of Tunisia comparing cockles non-parasitized (NP) and parasitized (P) with digenean parasites. Elements concentration in sediments analyzed in both sites revealed that Zinc (Zn), Chromium (Cr), and Lead (Pb) were the most abundant ones, while Cadmium (Cd) and Mercury (Hg) were less abundant. The bioaccumulation of trace metals and Arsenic (As) in the tissues of cockles seems to be modulated by both the infection state and the parasite species. The relationship between bioaccumulation of metals and As, trematode species and abiotic parameters showed that the availability of certain metals for uptake by P cockles of both sites was influenced by the salinity and temperature of the water. Our results corroborate the possibility of using digenean infecting bivalves in biomonitoring aquatic ecosystems.

Plasma Exchange for Lyme Neuroborreliosis delayed diagnosis: A case report.

Lyme neuroborreliosis (LNB) is a rare infectious disease, caused by Borrelia burgdorferi spirochetes and responsible for a variety of neurological manifestations. The most common manifestations of LNB in children are cranial nerve involvement, especially facial nerve palsy often accompanied by lymphocytic meningitis. In this article, we present a case of a 4-year-old boy presented to our emergency department with abdominal pain evolving for a week and symmetrical ascending progression of weakness responsible for severe respiratory failure. Diagnosis of Guillain-Barré syndrome (GBS) was initially suspected. Although our patient had received 2 courses (each of 5 days) of Intravenous immunoglobulin (IVG) treatment, no clinical improvement was observed. The diagnosis of LNB was confirmed by detection of both IgG and IgM specific antibodies in serum. The patient's muscle weakness got better after a 2- week course of Ceftriaxone but respiratory muscle failure didn't improve with two extubation failures. Consequently, we decided to conduct plasmapheresis procedures. We managed to extubate the child and discharge him after a good recovery of his symptoms. Pediatricians must consider LNB disease in the differential diagnosis of GBS, especially when the patient didn't recover after IVG treatment. This case shows that plasmapheresis could be effective for pediatric neuroborreliosis cases with severe neurological disorders.

Clinical, laboratory and evolutionary features of abdominal tuberculosis in comparison with other forms of extrapulmonary tuberculosis.

BACKGROUND/OBJECTIVES: Tuberculosis is a multisystem disease that might affect any organ. Abdominal tuberculosis (ABT) represents 5-17% from all extrapulmonary tuberculosis (EPT) sites. We aimed to study the clinical, laboratory and evolutionary features of ABT cases and to identify predictive factors associated with ABT. METHODS: We conducted a retrospective study including all patients hospitalized in the infectious diseases department for EPT between 1991 and 2019. We studied the characteristics of ABT cases, and we compared them with other EPT cases. RESULTS: We identified 519 patients with EPT, among whom 86 (16.6%) patients had ABT. There were 58 females (67.4%). Peritoneal tuberculosis was the most common clinical form of ABT (68.6%), followed by intestinal tuberculosis (18.6%). Patients aged 60 years and above were significantly less affected with ABT (odds ratio (OR) = 0.2; p = 0.001). The revealing systemic symptoms including fever (OR = 2.04; p = 0.006), weight loss (OR = 2.5; p < 0.001) and anorexia (OR = 1.7; p = 0.021) were significantly more frequent among ABT patients. Inflammatory markers including C-reactive protein levels (37 [10-89] mg/l vs 10 [4-57] mg/l; p < 0.001) and erythrocyte sedimentation rates (43 [15-95] mm/h vs 27 [15-60] mm/h; p = 0.044) were significantly higher among ABT cases. Multivariate logistic regression analysis showed that anorexia (adjusted OR (AOR) = 1.9; p = 0.015) and pulmonary involvement (AOR = 3.3; p = 0.002) were independent predictors of higher rate of ABT. Concomitant involvement of neuro-meningeal (AOR = 0.18; p = 0.001) and osteo-articular (AOR = 0.2; p = 0.01) sites, 40-59 (AOR = 0.2; p < 0.001) and ≥60 (AOR = 0.2; p < 0.001) age groups as well as hemoglobin rate (AOR = 0.7; p < 0.001) were independently associated with lower rate of ABT. CONCLUSIONS: Anorexia and pulmonary involvement were independent predictors of higher rate of ABT. Concomitant involvement of neuro-meningeal and osteo-articular sites, 40-59 and ≥60 age groups and hemoglobin rate were independently associated with lower rate of ABT.

Evaluation of guidelines for management of familial adenomatous polyposis in a multicenter pediatric cohort.

OBJECTIVE: To retrospectively assess, in a pediatric multicenter cohort, guidelines for the management of familial adenomatous polyposis (FAP). METHODS: Ten centers from the French-speaking Pediatric Gastroenterology Hepatology and Nutrition Group provided follow-up data on patients up to 18 years of age. Clinical records, genetic test results, endoscopy with histopathology examination, and therapeutic modalities were reviewed. RESULTS: A total of 70 children from 47 families were included. When initial consultation resulted from a surveillance program because of an affected family member, 12 of 59 children were already symptomatic. Among 11 patients whose initial consultation was based only on symptoms, families were unaware at the time of a familial FAP history for 7 children, whereas only 4 cases were sporadic. A panel of 27 different pathogenic adenomatous polyposis coli (APC) germ-line mutations and large genomic deletions were identified in 43 families. Extracolonic manifestations were found in half of the patients. As part of the standard practice for initial screening, the entire cohort underwent colonoscopy, which revealed adenoma above an intact rectosigmoid in 8 cases. Prophylactic colectomy was performed in 42 cases; high-grade dysplastic adenoma and 1 invasive carcinoma were detected in 6 children. For timing of surgery, indications were in accordance with recent international guidelines. CONCLUSIONS: Defining optimal screening and therapeutic modalities in pediatric FAP cohorts is a challenge. Specific advice for genetic screening, endoscopy surveillance, and type of surgery based on recent guidelines is recommended.

Antinuclear antibodies in children: clinical signification and diagnosis utility.

BACKGROUND: Antinuclear antibodies (ANA) test is used to screen adults as well as children for connective tissue diseases (CTD) and autoimmune hepatitis. However, interpretation of ANA positivity can be delicate. AIM: to determine clinical significance and diagnosis utility of ANA positivity in children. METHODS: Patients from a general pediatric department with ANA positive results were included (follow-up period of 2 years). ANA screening was performed by indirect immunofluorescence (IIF) on HEp-2 cells substrate (BioSystems®). In case of ANA positivity (cut-off: 1:80), the specificity was determined by IIF on Crithidia luciliea substrate (BioSystems®) and immunodot (Euroimmun®). RESULTS: Among 102 ANA tests, 55 (53,9%) were positive. We recorded the data of 38 patients (age average: 9,5 years - sex ratio: 0.72). The most frequent signs were join pain (55,3%). ANA titer varied between 1:80 (39,5% of cases) and 1:1280 (2,6% of cases). Typing was negative in 89,5% of cases. The majority (42,1%) of children with positive ANA test had musculoskeletal diseases. The others (57,9%) had systemic lupus erythematosus(n=2),  overlap syndrome(n=1), rheumatoid purpura(n=2), idiopathic thrombocytopenic purpura(n=1), coeliac disease(n=1) or non-autoimmune diseases/no confirmed diagnosis(n=15). CONCLUSIONS: ANA prevalence in children was relatively high. When the pretest probability is low, the positive predictive value for CTD or autoimmune hepatitis is low. However, depending on the clinical context, ANA detection can represent a supplement diagnostic tool for these diseases and/or can lead to a clinico-biological monitoring.

Characterization of a novel ABCC2 mutation in infantile Dubin Johnson syndrome.

BACKGROUND AND AIMS: The Dubin Johnson Syndrome (DJS) occurs mostly in young adults but an early-onset of the disease has been reported in less common forms (Neonatal DJS and Infantile DJS). In this case, the clinical findings are of limit for the DJS diagnosis. Hence, the genetic testing remains the method of choice to provide an accurate diagnosis. In our study, we aimed to perform a genetic analysis for two siblings presented with an intrahepatic cholestasis before the age of 1 year to provide a molecular explanation for the developed phenotype. PATIENTS & METHODS: A Tunisian family, having two siblings, manifesting signs of a hepatopathy, was enrolled in our study. A molecular analysis was performed, using a panel-based next generation sequencing, supplying results that were the subject of computational analysis. Then, a clinical follow-up was carried out to assess the evolution of the disease. RESULTS: The genetic analysis revealed the presence of a novel missense c.4179G > T, (p.M1393I) mutation in ABCC2 gene associated with a substitution c.2789G > A (R930Q) in ATP8B1 gene. Predictive results consolidated the pathogenic effect of both variants. These results confirmed the DJS diagnosis in the studied patients. The clinical course of both patients fit well with the benign nature of DJS. CONCLUSION: We described here a novel ABCC2 mutation associated with a putative ATP8B1 modifier variant. This finding constituted the first report of a complex genotype in DJS. Hence, genetic analysis by a panel-based next generation sequencing permits an accurate diagnosis and the identification of putative variants that could influence the developed phenotype.