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.

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.

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.

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.

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.

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.

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.

Epidemiology and disease burden of tuberculosis in south of Tunisia over a 22-year period: Current trends and future projections.

BACKGROUND: Tuberculosis (TB) is a public health problem worldwide. Characterizing its trends over time is a useful tool for decision-makers to assess the efficiency of TB control programs. We aimed to give an update on the current chronological trends of TB in Southern Tunisia from 1995 to 2016 and to estimate future trajectories of TB epidemic by 2030. METHODS: We retrospectively collected data of all notified TB new cases by the Center of Tuberculosis Control between 1995 and 2016 in South of Tunisia. Joinpoint Regression Analysis was performed to analyze chronological trends and annual percentage changes (APC) were estimated. RESULTS: In the past 22 years, a total of 2771 cases of TB were notified in Southern Tunisia. The annual incidence rate of TB was 13.91/100,000 population/year. There was a rise in all forms of TB incidence (APC = 1.63) and in extrapulmonary tuberculosis (EPTB) (APC = 2.04). The incidence of TB increased in children and adult females between 1995 and 2016 (APC = 4.48 and 2.37, respectively). The annual number of TB declined in urban districts between 2004 and 2016 (APC = -2.85). Lymph node TB cases increased (APC = 4.58), while annual number of urogenital TB decreased between 1995 and 2016 (APC = -3.38). Projected incidence rates would increase to 18.13 and 11.8/100,000 population in 2030 for global TB and EPTB, respectively. CONCLUSIONS: Our study highlighted a rise in all forms of TB and among high-risk groups, notably children, females and lymph node TB patients in the last two decades and up to the next one.

Philometra barbata n. sp. (Nematoda: Philometridae) from the red mullet Mullus barbatus (Perciformes, Mullidae) off Tunisia.

Helminthological examinations of the red mullet Mullus barbatus (Linnaeus) (Mullidae) from the Gulf of Hammamet, off Tunisia, revealed the presence of one undescribed gonad-infecting nematode species, Philometra barbata n. sp. (Philometridae). The new species as other congeneric species is mainly characterized by the shape of the caudal mound, the distribution of the caudal papillae and the shape of the gubernaculum with the presence of a dorsal protuberance consisting of two dorsolateral lamellar parts separated from each other by a smooth median field in the male. The new species differs from its gonadinfecting congeners in the body length of male, the length of spicules and gubernaculum. This is the second nominal species of Philometra reported from fishes of the family Mullidae and the 14th from the Mediterranean Sea.

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.

[Post-streptococcal glomerulonephritis in the south of Tunisia: A 12-year retrospective review]. / La glomérulonéphrite aiguë post-streptococcique de l'enfant dans le sud tunisien : étude rétrospective de 12 ans.

AIM: Post-streptococcal glomerulonephritis (PSGN) is a frequent cause of acute nephritis in children. This study aimed to describe the epidemiology, clinical characteristics and outcomes of PSGN and look for predictor's factors of severity. METHODS: A 12-year retrospective review of case notes and laboratory data was conducted at a department of pediatrics, pediatric emergency and intensive care, Hedi Chaker Hospital. RESULTS: One hundred seventy eight children were treated for PSGN with a mean age of 7.6 ans±3.43 ans. One hundred and forty-two patients (80%) had a history of a recent upper respiratory tract or skin infection. Streptococcal pharyngitis was the most common cause, identified in 113 patients (67.6%). Macroscopic hematuria and edema were noted in 135 (75.8%) and 114 cases (64%) respectively. Hypertension was present in 55 patients (31%). Oliguria was noted in 30 children (16.8%). Sixty-six subjects (37%) developed acute renal impairment (creatinine≥70 micromoles/L). No correlation was demonstrated between acute renal impairment and age, sex, triggering infection, anemia and white blood cell count. Creatinine greater than 56.35 micromoles/L was associated with a high risk of developing high blood pressure. The mean length of admission was 5.8 days±4.44. Only one subject has ongoing renal dysfunction. CONCLUSION: PSGN remains a common nephropathy in our region. The detection and effective treatment of any infection that may be involved can reduce the incidence of this disease.

[Positivity of antineutrophil cytoplasmic antibodies in children: prevalence and etiologies]. / Positivité des anticorps anti-cytoplasme des polynucléaires neutrophiles chez l'enfant : prévalence et étiologies.

In adults, anti-neutrophil cytoplasmic antibodies (ANCA) are considered as serological markers of several diseases, especially vasculitis and glomerulonephritis. Since ANCA are rarely positive in children, few data about the clinical relevance of these auto-antibodies in pediatric population have been reported. Therefore, our study aims to describe the spectrum of disorders associated with positive ANCA in Tunisian children. This study had been carried out over a period of 12 years and a half. All patients under the age of 15 for whom ANCA screening was performed in our laboratory were included. Clinical data were collected retrospectively. Indirect immunofluorescence (IFI) technique for ANCA detection was performed using PNN smears fixed with ethanol, formalin and, if necessary, methanol. Positive results were tested using immunodot to characterize the antigenic targets (myeloperoxydase (MPO) and proteinase 3 (PR3)). Our results showed that 410/5,990 (6.8%) laboratory requests for ANCA screening were for children. Forty (9.7%) requests were positive (24 children). Clinical data were available for 19 patients only. Sex-ratio (F/M) was 1.25. The mean age was 9 years and a half (3-15 years). The most frequent IIF patterns were x-ANCA (n=12) and p-ANCA (n=7). In our patients, the most frequent conditions associated to ANCA were treatment with benzylthiouracil for hypothyroidism (n=6), inflammatory bowel disease (n=4) and hemolytic anemia (n=4). In conclusion, the positivity of ANCA in children seems to be a rare event. Associated conditions include clinical disorders specific to the pediatric population. Treatment with benzylthiouracil is an etiology to be taken into consideration.

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.

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.

Protective effect of Sargussum vulgare sulfated polysaccharide against molecular, biochemical and histopathological damage caused by alloxan in experimental diabetic rats.

Diabetes is a serious condition that is linked to the development of oxidative stress. In the context of enhancing the biodiversity of Tunisia's flora, this study aimed to evaluate the effect of Sargussum vulgare sulfated polysaccharide (SVSP) on hyperglycemia and diabetes complications in the alloxan-induced diabetic rats. Our results showed a disturbance of carbohydrate, lipid, hematological and histopathological parameters, an increase in the α-amylase enzyme activity and damage to the pancreatic, hepatic and renal tissues in rats rendered diabetic by alloxan. In contrast, treatment with SVSP resulted in a correction of fasting and postprandial blood glucose and HbA1c through inhibition of pancreatic α-amylase. We also noticed an improvement in hemogram parameters and an attenuation of the pancreatic oxidative stress markers as well as histological protections. On the other hand, the administration of SVSP to diabetic rats caused, besides the correction of glycemic and lipid parameters, a good defense against hepatotoxicity and nephrotoxicity of diabetes as well as good antioxidant status and histological protections.