Cytokine and chemokine profiles in the sera of COVID-19 patients with different stages of severity.

INTRODUCTION: COVID-19 is a viral infection that disturbs the host's immune system and causes an overproduction of cytokines leading to a cytokine storm. The present study aimed to evaluate the serum levels of 27 protein biomarkers to determine their association with COVID-19 disease severity. METHODS: The serum levels of 89 patients with different degrees of COVID-19 disease severity [asymptomatic (n = 14), moderate (n = 14), severe (n = 30), and critical (n = 31)] and 14 healthy individuals were tested for a panel of 27 cytokines and chemokines using Luminex assay (27 Bio­Plex Pro Human Cytokine, Bio-rad™). RESULTS: IL-12, IL-2 and IL-13, as well as IL-17 and GM-CSF were clearly undetectable in asymptomatic patients. IL-8 levels were higher in asymptomatic compared with other groups. Very high levels of IL-6, IL-10 and the chemokines MIP-1α, MCP-1 and IP10 were associated with disease progression, while IL-4 tends to decrease with disease severity. CONCLUSION: Our study provides more evidence that excessive cytokine synthesis is linked to the disease progression.

Urine-based detection of HPV for cervical cancer screening: Time for standardized tests.

Cervical cancer is preventable because it has an established etiology, mainly attributed to a detectable pathogen, human papillomavirus (HPV). In 2018, the world health organization issued an unprecedented call for global action to eliminate cervical cancer by 2030. The adaptation of regular screening programs is fundamental to achieve the goal of cervical cancer elimination. However, it is still difficult to achieve satisfactory coverage rates of screening in developing countries as well as in developed countries because many women are reluctant to participate in gynecologic examination. HPV detection in urine is a convenient, widely acceptable by women and relatively affordable without the necessity for clinical visits to improve the coverage rates of cervical cancer screening. Unfortunately, the clinical implementation of urine-based tests for HPV detection has been hindered by the lack of standardized tests. Further optimization of protocols and standardization of urinary HPV detection are expected to be realized. With the advantages of urine sampling to overcome cost, personal, and cultural barriers, time has come for the standardized tests to facilitate a wide clinical implementation of urinary HPV detection that will significantly contribute to the WHO's goal, that is, to eliminate the cervical cancer globally.

Molecular evaluation of human papillomavirus as an oncogenic biomarker in prostate cancer.

BACKGROUND: Prostate cancer (PCa) is one of the most common tumors in men, regardless of ethnicity and demographics. In many risk factors causing PCa, genes and viral infections are strong candidates for the development of prostate tumors. Indeed, tissue infections of PCa have been reported by the presence of several types of viruses including Human Papillomaviruses (HPV). OBJECTIVE: the present study was planned to determine whether HPV DNA could be detected in the blood of known men with prostate cancer and to assess the potential association between HPV infection and clinico-pathological characteristics of the patients. MATERIALS AND METHODS: In order to achieve our objectives, 150 liquid blood samples were taken from Moroccan patients, 100 patients with PCa, and 50 control cases. The viral DNA was extracted, calibrated and the target genes were amplified by PCR using specific primers and the use of 2% agarose gel with visualization under UV. RESULTS: Of the 100 samples tested, (10%) were infected with HPV), However, none of the control cases were infected with HPV. The analysis of the data made it possible to establish a correlation between the frequency of the viral infection of the human papilloma and the tumoral criteria. CONCLUSION: Therefore, this study strengthens the potential role of HPV as a cofactor in prostate cancer development, and we propose that infection with this virus may be involved in the development of PCa metastases.

Biochemical responses in mice induced by saxitoxins extracted from the cockles Acanthocardia tuberculatum.

Harmful algae blooms have increased in frequency and geographic range in recent decades, and they produce toxins strains such as saxitoxins (STXs). they block voltage-gated sodium channels and can lead to several poisonings and the death of organisms that pose a significant risk to public and environmental health. The study of STXs toxicity has been carried out but little is known about the response of antioxidant enzymes activities to STXs in mice. The purpose of this study was to evaluate biochemical responses and oxidative stress induced by STXs extracted from Acanthocardia tuberculatum. To this end, daily, mice were treated orally for 7 days with sublethal concentrations (10 mg/100 g mouse). The animal's liver was assessed using biomarkers such as activities of catalase (CAT), thiobarbituric acid reactive substances (TBARS), glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and succinate dehydrogenase (SDH). In the blood, plasmatic markers were analysed as glutamic oxalic transaminase (GOT), glutamic pyruvic transaminase (GPT), creatinine phosphokinase (CPK), lactate dehydrogenase (LDH), urea and creatinine. Globally, test toxicity test showed a significant decrease in the weight at 10 mg /100 g mouse, and the results showed an increase of GPT, GOT, CPK, LDH, CAT and TBARS activities and the inhibitory effect of GAPDH activities but creatinine, urea and SDH activities showed no significative difference from the control. We concluded that STXs induce oxidative stress breaking in mice the balance of the defence system and causing oxidations reactions. Moreover, STXs affect energy metabolism in mice, however, renal function in mice is not affected by exposure to STXs.

Hepatitis A virus detection by RT-qPCR in shellfish samples from three Moroccan Atlantic coastal areas: Dakhla, Oualidia, and Moulay Bousselham.

This comparative study assessed hepatitis A virus (HAV) contamination in shellfish harvesting areas in Morocco, and the correlations between viral contamination and rainfall. To this aim, HAV contamination was evaluated in 156 shellfish samples collected at three Moroccan coastal areas (52 samples/area): Dakhla (class A), Oualidia (class B), and Moulay Bousselham (class C). Samples were collected monthly between March 2018 and March 2019, and included oysters from different farms at the Oualidia and Dakhla coastal areas, and wild mussels at the Moulay Bousselham lagoon. HAV was detected by RT-qPCR in 24/156 (15.38%) samples: 16/145 (11.03%) mussel (Mytillus galloprovincialis) and 8/156 (5.13%) oyster (Crassostea gigas) samples. The 16 mussel-positive samples (16/52) were collected at Moulay Bousselham, and the eight oyster-positive samples (8/52) were collected at Oualidia, and none at Dakhla (class A). The highest HAV contamination rate was recorded at Moulay Bousselham lagoon (class C) (30.77%), while it did not exceed 8% at Oualidia (class A). A nonsignificant, positive correlation between HAV-positive samples and mean rainfall was observed. The important HAV presence in bivalve mollusks from the Oualidia and Moulay Bousselham lagoons indicate the need of viral surveillance of bivalve mollusks and of improving wastewater quality to enhance shellfish safety.

Prevalence of specific and recurrent/founder pathogenic variants in BRCA genes in breast and ovarian cancer in North Africa.

BACKGROUND: Elucidation of specific and recurrent/founder pathogenic variants (PVs) in BRCA (BRCA1 and BRCA2) genes can make the genetic testing, for breast cancer (BC) and/or ovarian cancer (OC), affordable for developing nations. METHODS: To establish the knowledge about BRCA PVs and to determine the prevalence of the specific and recurrent/founder variants in BRCA genes in BC and/or OC women in North Africa, a systematic review was conducted in Morocco, Algeria, and Tunisia. RESULTS: Search of the databases yielded 25 relevant references, including eleven studies in Morocco, five in Algeria, and nine in Tunisia. Overall, 15 studies investigated both BRCA1 and BRCA2 genes, four studies examined the entire coding region of the BRCA1 gene, and six studies in which the analysis was limited to a few BRCA1 and/or BRCA2 exons. Overall, 76 PVs (44 in BRCA1 and32 in BRCA2) were identified in 196 BC and/or OC patients (129 BRCA1 and 67 BRCA2 carriers). Eighteen of the 76 (23.7%) PVs [10/44 (22.7%) in BRCA1 and 8/32 (25%) in BRCA2] were reported for the first time and considered to be novel PVs. Among those identified as unlikely to be of North African origin, the BRCA1 c.68_69del and BRCA1 c.5266dupC Jewish founder alleles and PVs that have been reported as recurrent/founder variants in European populations (ex: BRCA1 c.181T>G, BRCA1 c1016dupA). The most well characterized PVs are four in BRCA1 gene [c.211dupA (14.7%), c.798_799detTT (14%), c.5266dup (8.5%), c.5309G>T (7.8%), c.3279delC (4.7%)] and one in BRCA2 [c.1310_1313detAAGA (38.9%)]. The c.211dupA and c.5309G>T PVs were identified as specific founder variants in Tunisia and Morocco, accounting for 35.2% (19/54) and 20.4% (10/49) of total established BRCA1 PVs, respectively. c.798_799delTT variant was identified in 14% (18/129) of all BRCA1 North African carriers, suggesting a founder allele. A broad spectrum of recurrent variants including BRCA1 3279delC, BRCA1 c.5266dup and BRCA2 c.1310_1313detAAGA was detected in 42 patients. BRCA1 founder variants explain around 36.4% (47/129) of BC and outnumber BRCA2 founder variants by a ratio of ≈3:1. CONCLUSIONS: Testing BC and/or OC patients for the panel of specific and recurrent/founder PVs might be the most cost-effective molecular diagnosis strategy.

Impact evidence of the vitamin D status in hypertensive patients in Morocco.

Vitamin D deficiency is a hot topic as it affects about 50% of the world's population. Beyond its extra-osseous effects, notably on the cardiovascular system. In recent years many epidemiological studies have looked at the role of vitamin D status in modulating blood pressure. It appears that there is an inverse association between serum vitamin D levels and blood pressure. It is in this context that our prospective study on the Moroccan population allows having global knowledge of the association between vitamin D and arterial hypertension. This is a descriptive study of vitamin D status in Moroccan hypertensive patients in whom blood samples were taken for serum 25(OH)D determination. The data collected were anonymised and entered into SPSS software. X2 and ANOVA tests were used to investigate the existence of correlations between vitamin D status and age, age at menopause, body mass index, sex and hypertension.  1015 hypertensive patients were included in this study with a female predominance of 84.2%, mainly over 50 years old in 80.8%, of whom 31.5% were overweight or obese, 13.2% had a hypertensive parent and 70.8% had vitamin D deficiency. Vitamin D regulates at least 3% of the human genome with receptors throughout the body, including vascular smooth muscle cells, vascular endothelium and cardiomyocytes, where it acts by vasodilation and by blocking the renin-angiotensin-aldosterone system (RAAS) to lower blood pressure. There is a relationship between low serum vitamin D levels and high blood pressure. Our study concluded that there is an association between hypovitaminosis D and the pathology of hypertension. However, further randomised studies are needed and in the meantime, clinicians could propose it in the therapeutic arsenal of Moroccan hypertensive patients.

Molecular characterization of rotavirus in oysters from Oualidia lagoon in Morocco.

Group A rotavirus (RVA) is the most important foodborne pathogen and is transmitted by oysters. The purpose of this study was to monitor and evaluate the prevalence of RVA in oysters collected in Oualidia Lagoon, Morocco from March 2018 to March 2019 and its relationship to watershed contribution. To achieve this, 312 oysters from 26 lots were collected and tested for viral contamination using real-time polymerase chain reaction. The results showed that 46.15% of the tested samples were positive for RVA. The seasonal distribution of RVA was statistically significant. This is a warning signal against the potential health risks in this oyster growing area, following the consumption of oysters contaminated with RVA, which presents a huge threat to human health resulting in serious illnesses of gastroenteritis.

Human papilloma virus profiles in breast cancer in correlation with Vitamin D.

The major roles of vitamin D in the genesis of breast cancer and as an immunomodulator against acute and chronic infections have been the subject of much research in recent years. A low vitamin D status could decrease the function of blocking the cell multiplication cycle of the cancer process and weaken the immune system. In this context, we were interested in the implication of vitamin D status in women with human papilloma virus (HPV)-induced breast cancer. Our study included 63 women, 53 with breast cancer and 10 healthy women, and we measured the plasma 25(OH)D3 level and looked for the presence of HPV by PCR in our population. 90.6% had low serum 25(OH)D3 levels and HPV was found in 41% of cases. In this regard, the data in the literature are discordant. Vitamin D status could explain the concomitance of the two conditions, breast cancer and HPV; it would be desirable to broaden the sample in order to better define its impact.

Detection of hypermethylation BRCA1/2 gene promoter in breast tumours among Moroccan women.

BACKGROUND: The promoter region is a key element of gene expression regulation. In mammals, most of the genes present, at the level of their promoter, a large number of islands CpG. Age also is seen as another factor for developing breast cell cancer reaching the tumour stage. AIM: This study aimed to explore the hypermethylation of the BRCA1/2 promoter gene in women breast cancer and correlation with age and tumour stage. MATERIALS AND METHODS: Fifty biopsies were derived from Moroccan women treated for breast carcinoma, the DNA extracted was treated by bisulphite and the targeted BRCA1/2 Amplicons were amplified by specific methylation primers (MSP). RESULTS: The result shows that 62% of the samples were BRCA1 methylated in addition and negative result for BRCA2, these positive epigenetic factor were remarkable in women over 47 years and at the stage of malignant tumour. CONCLUSION: These results show that half of the methylated samples are positive with a majority of over 47 years old, and confirms that age might be an additional factor for breast cancer development.

A computational approach to the study of interactions between proteins and miR10-b, miR-335, and miR-21 involved in breast cancer.

MiR-10b, miR-335, and miR-21 are classes of microRNAs (miRNAs) that are overexpressed in breast cancer. Thus, in our study we aimed to test the hypothesis that miRNAs may have direct interactions with proteins and the possibility to inhibit/activate the functional site of proteins and enzymes. For this purpose, we choose three miRNAs involved in breast cancer to study interactions between some proteins and genes, including BRCA1 and PTEN, by processing the docking and matching tools using the Hex8 and HADDOCK server. Mathematically, the hidden Markov models were created by using MATLAB script according to the algorithm in order to study and validate the interactions and bonds between proteins and miRNAs. The main results demonstrate the ability of miR-10b, miR-335, and miR-21 to create direct interactions with 3D protein structures. Furthermore, these results may lead to another pathway of research, i.e. the direct interaction between proteins and their sub-units, to highlight the data obtained previously and demonstrate that proteins may directly interact with ncRNA instead of mRNA. Moreover, our study suggests developing research on different pathways of association proteins-miRNAs as a part of epigenetic extra-nuclear regulation. Taken together, our study provides the first evidence of direct interactions between miRNAs and proteins.

Human papillomavirus genotypes distribution by cervical cytologic status among women attending the General Hospital of Loandjili, Pointe-Noire, Southwest Congo (Brazzaville).

HPV infection is associated with cervical cancer, one of the major public health problems in developing countries. In the Republic of Congo, despite of the high age-standardized incidence rate estimated at 25.2 per 100,000 women, molecular epidemiology data on HPV infections are very limited. We investigated HPV genotypes distribution in cervical smears among patients attending the General Hospital of Loandjili, Southwest Congo. A cross-sectional hospital-based study was conducted on 321 women. Liquid-based cytology samples were collected for cytological diagnosis and HPV detection. Nested-PCR was performed using MY09/MY11 and GP5+/GP6+ primers with genotyping by direct sequencing. Type-specific PCR for HPV-6, -11, -16, -18, -31 and -33 was also used to assess multiple infections. Out of 321 women examined, 189 (58.8%) had normal cytology, 16 (5.0%) had ASCUS and 116 (36.1%) had cytological abnormalities. HPV-DNA was detected in 22 (11.6%), 6 (37.5%), and 104 (89.6%) normal cytology, ASCUS and cytological abnormalities respectively. HPV16 was the most prevalent genotype regardless of cytological status followed by HPV70 in women without lesions and HPV33 among those with lesions. HR-HPV prevalence varied significantly according to the cervical cytology (P = 0.000). Among women without lesions, two peaks of HPV infections were observed in age group less than 30 years (60.0%) and in age group 50-59 years (7.1%). Age, age of first sex, multiple sexual partners and pregnancies were the risk factors for HPV infection in women without lesions. Our findings could be used as evidence data base for future epidemiological monitoring in this region.

A sheeppox outbreak in Morocco: isolation and identification of virus responsible for the new clinical form of disease.

BACKGROUND: Sheeppoxvirus (SPPV) is a member of the Capripoxvirus genus of the Poxviridae family, which causes significant economic losses in Morocco. The resurgence of the sheeppox disease during 2010 was characterized by an emergence of a classical nodular form for the first time in Morocco. However, little is known about the virus strain responsible for nodular form. In this study, thirty three sheep, from the eastern region of Morocco, clinically infected were examined and dead animals were autopsied.A rapid diagnostic assay for SPPV using different type of clinical samples would be useful for outbreak management. The aim of this work was to isolate the virus strain responsible for nodular form and we identified and compared by phylogenetic analysis the field strain with Moroccan vaccine strain targeting the thymidine kinase (TK) gene and the chemokine analogue receptor of interleukin (IL8) gene. Further, it was important to investigate and validate a real-time PCR using different clinical and post-mortem samples to manage epidemic sheeppox disease. RESULTS: The nodular form of sheeppox disease observed in Morocco was clinically characterized by fever, depression, lacrimation, diarrhea in lambs and nodule. At necropsy, the most affected organ was the lung. The etiological strain was successfully isolated from lung nodule in a dead lamb and was identified by using real-time PCR that has been tested and validated on different types of clinical and post mortem samples from naturally infected animals. Sequence and phylogenetic analysis of TK and IL8 gene showed that there was a very close relationship between field and vaccine strain. They were clustered within other SPPV strains. CONCLUSION: In the current study, we show for the first time the nodular form of sheeppox in Morocco. We demonstrate a robust real-time PCR-based diagnostic assay to detect the sheeppox virus in multiple sample that can be implemented to efficiently manage the disease outbreak. Our study also offers the prospect for future molecular studies to understand the clinical forms.

Specific genotypes of human papillomavirus in 125 high-grade squamous lesions and invasive cervical cancer cases from Congolese women.

BACKGROUND: Knowledge on HPV prevalence and genotype distribution in HSIL and ICC is highly essential for the introduction of an effective vaccination program and appropriate epidemiological monitoring of viral ecology before and after vaccination in Congo. This study aimed to determine the specific-HPV genotypes in HSIL and ICC among women in southwestern Congo. METHODS: 125 archival paraffin-embedded biopsy collected between 2008 and 2012 and histologically diagnosed were investigated. DNA extraction was performed using the phenol/chloroform method. HPV search was performed by nested-PCR using MY09/MY11 and GP5+/GP6+ consensus primers followed by direct sequencing. RESULTS: The mean age of participants was 44.3 ± 8.2 years. Overall, HPV prevalence was 89.6% (112/125) with all high-risk genotypes. HPV-DNA was detected in 81.5% (53/65) of HSIL and 98.3% (59/60) of ICC. HPV 16 the most common genotype was detected in 47.1% (25/53) of HSIL and 52.5% (31/59) of ICC. Other types identified were: HPV 33 (22.6%), HPV 18 (15%), HPV 31 (11.3%) and HPV 69 (3.7%) in HSIL, and HPV 33 (28.8%), HPV 18 (11.8%), HPV 31 (5%) and HPV 35 (1.7%) in ICC. Knowing that the ADC accounted for 6.7% (4/60) of ICC cases, HPV 18 was identified in 25% (1/4) of these cases against 75% (3/4) for HPV 16. CONCLUSION: Our study showed that HPV 16, 33, 18 and 31 were the four most common genotypes in women with HSIL and ICC. These findings indicate that current vaccines against HPV could help to reduce the burden of cervical cancer in Congo.

Prevalence of co-infection with human papillomavirus and Chlamydia trachomatis and risk factors associated with cervical cancer in Congolese women.

The human papillomavirus (HPV) is one of the most frequently diagnosed viruses in developing countries. Chlamydia trachomatis (CT) is an important cofactor in HPV-induced cervical cancer. Cervico-uterine smears were taken for cytology, and a total of 131 samples were analysed. HPV prevalence and CT were detected using specific primers (L1 gene and omp-1 gene). 23 (17.5 %) HPV-only samples were detected, CT-only positives were 10 (7.6 %). And HPV/CT co-infection was 13 (9.9 %). Identified risk factors associated with HPV/CT co-infection were risky sexual behaviour and cytology status. The prevalence of HPV and CT and their co-infection rates being high in our study population, may be an indicator of cervical cancer risk. Consequently, there is an urgent need to raise awareness and take appropriate precautions.

An easy, simple inexpensive test for the specific detection of Pectobacterium carotovorum subsp. carotovorum based on sequence analysis of the pmrA gene.

BACKGROUND: The species Pectobacterium carotovorum includes a diverse subspecies of bacteria that cause disease on a wide variety of plants. In Morocco, approximately 95% of the P. carotovorum isolates from potato plants with tuber soft rot are P. carotovorum subsp. carotovorum. However, identification of this pathogen is not always related to visual disease symptoms. This is especially true when different pathogen cause similar diseases on potato, citing as an example, P. carotovorum, P. atrosepticum and P. wasabiae. Numerous conventional methods were used to characterize Pectobacterium spp., including biochemical assays, specific PCR-based tests, and construction of phylogenetic trees by using gene sequences. In this study, an alternative method is presented using a gene linked to pathogenicity, in order to allow accuracy at subspecies level. The pmrA gene (response regulator) has been used for identification and analysis of the relationships among twenty nine Pectobacterium carotovorum subsp. carotovorum and other Pectobacterium subspecies. RESULTS: Phylogenetic analyses of pmrA sequences compared to ERIC-PCR and 16S rDNA sequencing, demonstrated that there is considerable genetic diversity in P. carotovorum subsp. carotovorum strains, which can be divided into two distinct groups within the same clade. CONCLUSIONS: pmrA sequence analysis is likely to be a reliable tool to identify the subspecies Pectobacterium carotovorum subsp. carotovorum and estimate their genetic diversity.

Detection of HPV in urine for cervical cancer screening: Feasibilty of an assay system.

The detection of urine HPV is considered as a promosing alternative to increase the screening coverage of cervical cancer. However, the validated assay of urine HPV is still scarse. We described a nouvel assay syetem for the urine-based detection of HPV in the framework of HPV screening. This sytsem consisted of Automate Nimbus extraction of DNA and Anyplex™ II HPV HR Detection PCR of HPV DNA. We validated this system by spiking HPV-infected cervical cancer cell line HeLa cells into normal urine and compared the prelimary results of cervical samples and urine samples. We found that this system could detect as few as 5 HeLa cells in normal urine model. Some discordances of HPV results between cervical samples and urine samples were observed. We concluded that this assay system could be applied for the detection of HPV in urine. A large scale study is necessary to evaluate the clinical significance of this assay system.

Germline mutations of B-Raf proto-oncogene and pathological implications in prostate cancer: observational study.

B-Raf proto-oncogene has been found in a variety of neoplasms. BRAF stimulation can promote tumour proliferation through the activation of the MAP/ERK kinase pathway. This study aimed to determine the germline spectra of BRAF and the association with pathological criteria of prostate tumours. Methods: Fifty blood samples from men treated with prostate cancer were analyzed for BRAF germline mutations and confirmed by Sanger sequencing, in addition, to establishing the frequencies and clinical correlations of frequent mutations in the BRAF gene for both exon 11 and exon 15. The frequency and distribution of high-frequency mutations were analyzed according to the pathological criteria of the patients. Results: Frameshift mutations: c.1628_1629insA and c.1624_1625insT with a frequency of (46%) and (18%), respectively, Nonsense mutations: c.1181C>A (p.Ser394Ter) was detected in one patient, missense mutations: c.1226A>G (p.Gln409Arg), c.1270T>C (p.Trp424Arg), c.1270_1271delins2 (p.Trp424Leu), with a frequency of (4%) were detected. There was no significant difference between mutation carriers and non-carriers regarding medical and surgical history, but prostate-specific antigen concentration was significantly different between the two groups. Conclusion: The results of this study elucidate the presence and involvement of germline mutations in prostate cancer, which could serve as a potential indicator for the diagnosis and therapeutic management of prostate cancer in the population studied.

Hepatitis A and E Viruses in Mussels from Cherrat Estuary in Morocco: Detection by Real-Time Reverse Transcription PCR Analysis.

The aim of the present study was to evaluate hepatitis A virus (HAV) and hepatitis E virus (HEV) contamination in mussels (Mytilus galloprovincialis) from Cherrat estuary (Moroccan Atlantic Coast), Morocco. In total, 52 samples (n = 12 mussels/each) were collected at four sites in the estuary, monthly, between March 2019 and March 2020. HAV and HEV were detected by real-time reverse transcription polymerase chain reaction (RT-PCR) according to the ISO/TS 15216 method. HAV was detected in 46.15% of analyzed samples. Conversely, HEV was not detected in any sample. Moreover, the HAV detection rate was significantly associated with seasonal rainfall variations. This qualitative study on HAV and HEV contamination highlights the interest of studying mussel samples from wild areas. As HAV presence in mussels represents a potential health risk, viral contamination surveillance of mussels is necessary to protect consumers. HAV shellfish contamination must be monitored at Cherrat estuary because of the role played by shellfish as HAV reservoirs and/or vehicles in fecal-oral HAV transmission.