Leveraging wastewater surveillance for managing the spread of SARS-CoV-2 and concerned pathogens during FIFA World Cup Qatar 2022.

Wastewater-based epidemiology (WBE) has been proven effective for the monitoring of infectious disease outbreaks during mass gathering events and for timely public health interventions. As part of Qatar's efforts to monitor and combat the spread of infectious diseases during the FIFA World Cup Qatar 2022™ (FWC'22), wastewater surveillance was used to monitor the spread of SARS-CoV-2, human enterovirus, and poliovirus. The screening covered five major wastewater treatment plants servicing the event locations between October 2022 and January 2023. Viruses were concentrated from the wastewater samples by PEG precipitation, followed by qRT-PCR to measure the viral load in the wastewater. As expected, SARS-CoV-2 and enterovirus RNA were detected in all samples, while poliovirus was not detected. The concentration of SARS-CoV-2 was correlated with population density, such as areas surrounding the World Cup venues, and with the number of reported clinical cases. Additionally, we observed temporal fluctuations in viral RNA concentrations, with peak levels coinciding with the group stage matches of the FWC'22. This study has been useful in providing public health authorities with an efficient and cost-effective surveillance system for potential infectious disease outbreaks during mega-events.

Seroprevalence and risk factors of Toxoplasma gondii infection in slaughtered chickens in Tripoli, Lebanon.

Toxoplasma gondii is a cosmopolitan protozoan parasite that has a wide range of intermediate hosts. It infects all warm-blooded animals, including humans and birds. The latter typically pick up the infection by ground feeding, and people can contract the parasite from eating undercooked chicken meat. In recent years, investigations into T. gondii infection in poultry have been reported worldwide. However, there is no epidemiological data regarding the seroprevalence of anti-T. gondii antibodies in chicken in Lebanon. Thus, the current investigation was carried out to determine the seroprevalence and associated risk factors of T. gondii infection in chicken destined for human consumption in the Tripoli district of Lebanon. For this, a cross-sectional study was carried out between April 2021 and February 2022. Blood samples were collected from 400 chickens in four poultry abattoirs in Tripoli. The modified agglutination test (MAT) was used to test sera for T. gondii antibodies. The association of T. gondii seroprevalence with potential risk factors was assessed using the Chi-square test. Multivariate analysis was used to confirm the association. The seroprevalence of T. gondii antibodies reported in this study was 13% (52/400); it was higher in the free-range chicken group (19.3%, 29/150) than in the caged group (9.2%, 23/250) (OR = 2.365; 95% CI: 1.311-4.267) (P = 0.004). The wet season and the presence of cats in the poultry farms were significantly associated with an increased seropositivity to T. gondii infection (P ≤ 0.0001). Given the occurrence of T. gondii antibodies in slaughtered chicken in this area, the consumption of raw or undercooked chicken meats may pose a serious threat to public health and highlight the need to implement appropriate precautionary strategies to halt the spread of T. gondii to humans.

[Déterminants de la qualité de vie liée à la santé chez les adolescents : le rôle de la diète méditerranéenne]. / Determinants of health-related quality of life among adolescents: The role of the Mediterranean diet.

BACKGROUND: A number of physical, emotional, cognitive and psychological changes shape the adolescence phase, which may influence health and well-being. Therefore, it is important to develop healthy eating habits that will persist into adulthood. OBJECTIVE: This study was conducted in order to investigate in depth the association between components of the Mediterranean diet (MD) and health-related quality of life (HRQoL), along with its different subscales, among Lebanese adolescent students attending schools in Tripoli, North Lebanon. METHODS: A cross-sectional survey was undertaken among 798 teenagers aged 11 to 18. All responded to a questionnaire that evaluated their HRQoL, sociodemographic characteristics and adherence to the Mediterranean diet. RESULTS: The results of the study revealed that only 15.9% of the students had a high adherence to the MD. With regard to the components of the MD associated with the HRQoL, it was revealed that fruit intake by the students (once daily) was associated with a better general well-being (P = 0.047). In addition, fish consumption (two to three times per week) and daily intake of a dairy product for breakfast were positively associated with the total KIDSCREEN score (P < 0.001 and P = 0.003, respectively). Finally, skipping breakfast was correlated with a poorer general well-being (P = 0.013). CONCLUSION: Our results suggest a strong correlation between MD adherence and HRQoL in Lebanese adolescents. However, future studies are warranted in order to strengthen the evidence of this association.

Parasitic Contamination of Fresh Leafy Green Vegetables Sold in Northern Lebanon.

Contaminated, raw or undercooked vegetables can transmit parasitic infections. Here, we investigated parasitic contamination of leafy green vegetables sold in local markets in the Tripoli district, Lebanon, during two consecutive autumn seasons (2020-2021). The study involved the microscopic examination of 300 samples of five different types of vegetables (60 samples per type) and used standardized qualitative parasitological techniques for some protozoa and helminths. The results showed that 16.7% (95% interval for p: 12.6%, 21.4%) (50/300) of the vegetable samples were contaminated with at least one parasite. The most frequently detected parasite was Blastocystis spp. (8.7%; 26/300); this was followed in frequency by Ascaris spp. (3.7%; 11/300). Among the different vegetable types, lettuce (23.3%; 14/60) was the most contaminated, while arugula was the least contaminated (11.7%; 7/60). The statistical analysis did not reveal any significant association between the prevalence of parasitic contamination and the investigated risk factors, which included collection date, vegetable type, market storage status, and wetness of vegetables at the time of purchase (p > 0.05). The high prevalence of parasitic contamination also suggested the potential presence of other microbial pathogens. These findings are important because leafy green vegetables are preferentially and heavily consumed raw in Lebanon. Thus, implementing effective measures that target the farm-to-fork continuum is recommended in order to reduce the spread of intestinal pathogens.

High seroprevalence of SARS-CoV-2 antibodies in household cats and dogs of Lebanon.

The COVID-19 pandemic has been declared in late 2019. It is caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Flu-like symptoms and acute respiratory illnesses are the main manifestations of the disease. Recent studies have confirmed the susceptibility of domestic animals to SARS-CoV-2 infection. However, the seroprevalence of SARS-CoV-2 in household pets and the importance of pets in the epidemiology of this infection remain unknown. In Lebanon, there is no epidemiological data regarding SARS-CoV-2 infection in companion animals. Thus, this investigation aimed to determine the seroprevalence of SARS-CoV-2 antibodies in household pets of Lebanon during the COVID-19 pandemic. A cross-sectional study was carried out between April 2020 and February 2021. Blood samples from 145 cats and 180 dogs were collected from 12 veterinary clinics located in the North, Mount, and Beirut governorates. A validated ELISA assay was used to detect the anti- SARS-CoV-2 in the sera of the tested animals. An overall seroprevalence of 16.92% (55/325) was reported; 13.79% seroprevalence was found in cats (20/145) and 19.44% (35/180) in dogs. The young age and the cold season were significantly associated with an increased seropositivity rate to SARS-CoV-2 infection (P < 0.01). These results confirm the circulation of SARS-CoV-2 in household pets, in various geographical regions in Lebanon. Although, there is a lack of evidence to suggest that naturally infected pets could transmit the SARS-CoV-2 infection. Yet, owners diagnosed with COVID-19 should limit their contact with their animals during the course of the disease to curb the risk of transmission.

Occurrence and risk factors of feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV) in cats of Lebanon.

A cross-sectional study was carried out between April 2020 and August 2021. Blood samples were collected from 260 household cats recruited in different clinics in North and Mount Lebanon, with the aim of determining the seroprevalence of feline immunodeficiency virus (FIV) and feline leukaemia virus (FeLV). The seroprevalence reported reached 18.84% (95% CI 0.14-0.24) and 13.84% (95% CI 0.09-0.18) for FIV and FeLV, respectively. FIV seropositivity was associated with a younger age, health and neuter status, lymphoma, lethargy, and vomiting. Furthermore, male sex, neuter status, lymphoma, anaemia, lethargy, and vomiting were significantly associated with FeLV seropositivity. This first data from Lebanon emphasizes the need for implementing preventive programmes to cope with FIV- and FeLV-associated morbidity and mortality among cats.

Seroprevalence and risk assessment of Toxoplasma gondii infection in sheep and goats in North and Beqaa governorates of Lebanon.

Background and Aim: Toxoplasmosis is a disease caused by the protozoan parasite Toxoplasma gondii that affects both humans and animals, leading to abortions and significant clinical manifestations in pregnant and immunocompromised hosts, in addition to massive economic losses in animal industries. Data from Lebanon are scarce regarding the seroprevalence of T. gondii infection in livestock. This study aimed to estimate the seroprevalence and assess the associated risk factors of T. gondii infection in sheep and goats in Lebanon. Materials and Methods: A cross-sectional study was carried out from May 2020 to April 2021. Blood samples from 150 sheep and 145 goats (total 295) destined for human consumption were obtained from 20 Lebanese farms located in the North and Beqaa governorates. The anti-T. gondii immunoglobulin G antibodies were assayed through means of a modified agglutination test with a cutoff titer of 20. Results: An overall seroprevalence of 48.5% (143/295) was reported: About 56.6% seroprevalence was found in sheep (85/150) and 40% (58/145) in goats. Adult age, female gender, and the wet season were significantly associated with an increased seropositivity rate of T. gondii infection (p < 0.001, p = 0.001, and p = 0.043, respectively). Conclusion: These results confirm the spread of T. gondii in sheep and goats destined for human consumption in various geographical regions in Lebanon. Therefore, continuous monitoring of T. gondii infection in livestock is warranted to control the spread of the infection and limit its potential transmission to humans through the consumption of raw or undercooked meat.

Association of Inherited Thrombophilia with Recurrent Pregnancy Loss in A Population of Lebanese Women: A Case Control Study.

Recurrent pregnancy loss (RPL) complication is a challenge of reproductive medicine due to its often unknown etiology.
A case-control study was carried out between June 2019 and April 2020 to examine the correlation between RPL
and inherited thrombophilia (IT), namely mutations in factor V Leiden (FVL G1691A), prothrombin (FII G20210A),
and methylenetetrahydrofolate reductase (MTHFR C677T). A total of 120 Lebanese women with RPL was studied
and compared, for the frequency of these mutations, to 100 healthy reproductive Lebanese women. The association
between the zygosity status of the three tested mutations, the existence of more than one prothrombotic single nucleotide
polymorphisms (SNPs), and the increased risk of RPL were examined using Chi-square or two-tailed fisher exact
test, and the student t test. The predictive factors of RPL were analyzed using a multiple logistic regression model.
P<0.05 was considered to be statistically significant. Our results showed statistically significant higher frequencies
of FVL G1691A and FII G20210A mutations among the cases with RPL compared to the control group. Thus, RPL is
associated with FVL G1691A and FII G20210A mutations. These mutations seem to increase the risk of RPL in the
Lebanese women. Therefore, we suggest thrombophilia screening and adequate genetic counseling for women with
RPL and at high-risk to plan for primary prevention, avoiding thromboembolic or obstetric accidents, and reducing
the associated morbidity and mortality among Lebanese women.

Incidence and Mortality of Extrapulmonary Tuberculosis in Ukraine: Analysis of National Surveillance Data.

BACKGROUND: Improved understanding of the epidemiology and mortality risk factors of extrapulmonary tuberculosis (EPTB) may facilitate successful diagnosis and management. METHODS: We analyzed national surveillance data from Ukraine to characterize EPTB subtypes (ie, localized in different anatomic sites). We calculated annual reported incidence, stratified by age, sex, and human immunodeficiency virus (HIV) status. Using Cox regression, we estimated mortality risk factors. RESULTS: Between January 2015 and November 2018, 14 062 adults/adolescents (≥15 years) and 417 children (<15 years) had EPTB with or without concomitant pulmonary TB. The most commonly reported EPTB subtypes were pleural, peripheral lymph node, and osteoarticular. Most EPTB subtype notifications peaked at age 30-39 years and were higher in males. In adults/adolescents, most peripheral TB lymphadenitis, central nervous system (CNS) TB, and abdominal TB occurred in those with untreated HIV. CNS TB notifications in people without HIV peaked before age 5 years. Adults/adolescents with CNS TB (adjusted hazard ratio [aHR]: 3.22; 95% CI: 2.89-3.60) and abdominal TB (aHR: 1.83; 95% CI: 1.59-2.11) were more likely to die than those with pulmonary TB. Children with CNS TB were more likely to die (aHR: 88.25; 95% CI: 43.49-179.10) than those with non-CNS TB. Among adults/adolescents, older age and HIV were associated with death. Rifampicin resistance was associated with mortality in pleural, peripheral lymph node, and CNS TB. CONCLUSIONS: We identified the most common EPTB subtypes by age and sex, patterns of EPTB disease by HIV status, and mortality risk factors. These findings can inform diagnosis and care for people with EPTB.

Understanding and Responding to Substance Use and Abuse in the Palestinian Refugee Camps in Lebanon Prior to and During COVID-19 Times.

Due to its geographical proximity to the Syrian conflict and the occupied territories, Lebanon has experienced an influx of refugees in recent times. Palestinian refugees are an identified key vulnerable population, with displaced communities increasingly experiencing camp insecurity, vulnerability to drug use and related health harms. A qualitative study consisting of in-depth interviews and focus group discussions (FGDs) was undertaken as part of a regional exercise investigating Palestinian community experiences of substance and drug use in refugee camps. Thematic analysis triangulated the perspectives of 11 professional stakeholders representing United Nations, human rights and non-governmental organizations (NGOs), and eight Palestinian community members. Emerging themes centered on the interplay between socio-economic instability, lack of law enforcement and camp governance contributing to concerning levels of familial, drug and camp violence, trafficking and availability of drugs. Transactional sex and the exploitation of women and children in drug dealing, diversification toward drug manufacture and dealing of drugs with the outside community were described. There is a lack of harm reduction and rehabilitation supports for those in need. This study highlights the complexities in tackling drug dealing and related criminal activity within refugee camps and humanitarian settings, and the vulnerabilities of those living within to harmful drug use.

Emergence of gram-negative organisms as the cause of infections in patients with sickle cell disease.

BACKGROUND: Patients with sickle cell disease are at higher risk of infections with encapsulated bacteria due to immature immune responses and functional asplenia. We aimed to study our patient population for the emergence of gram-negative organisms other than Salmonella as the cause of osteomyelitis and document a vast decrease in Streptococcus pneumoniae bacteremia rates. METHODS: We conducted a retrospective chart review of 158 patients with sickle cell disease registered at our hospital. Over a period of 13 years, every patient presenting to the emergency department (ED) with fever had their medical record reviewed for blood cultures, wound cultures, and magnetic resonance imaging results for osteomyelitis. RESULTS: The number of patients presenting to the ED with fever was 105, with 581 febrile episodes and 893 blood cultures. Among those, no culture grew Streptococcus pneumoniae, 14 grew coagulase-negative staphylococci (1.5%), one grew Salmonella enterica Paratyphi B, and three grew Salmonella enterica group C (in the same patient). The total number of osteomyelitis episodes in patients with sickle cell disease presenting with fever and documented by imaging was nine (1.5%). In patients with osteomyelitis, organisms were isolated in four patients (44%), including Enterobacter cloacae, Bacteroides, Pseudomonas aeruginosa, and Salmonella enterica group C. CONCLUSIONS: Immunization against Streptococcus pneumoniae and the use of prophylactic penicillin has virtually eliminated pneumococcal bacteremia among our patients. We observed the emergence of gram-negative organisms other than Salmonella as the cause of osteomyelitis in patients with sickle cell disease.

Inherited Thrombophilia in a Lebanese Family of Four Generations: A Case Report of Recurrent Miscarriage.

INTRODUCTION: Factor V Leiden (G1691A), prothrombin (G20210A) and MTHFR (C677T) gene mutations were investigated in many studies for their association with Deep Venous Thrombosis. CASE PRESENTATION: A North Lebanese family has been examined, from an index case, a 40-year-old woman, who had a history of venous thrombosis with unexplained recurrent miscarriage. The index case was found to be heterozygous for factor V Leiden G1691A, prothrombin G20210A, and methylenetetrahydrofolate reductase C677T gene variants. Her family members were heterozygous for at least two of the three-point mutations, and multiple risk factors associated with thrombophilia were identified. CONCLUSION: Our findings emphasize the need for clarifying the utility and futility of thrombophilia testing in the era of molecular diagnostics.

Relationship Between Pneumocystis carinii Burden and the Degree of Host Immunosuppression in an Airborne Transmission Experimental Model.

To quantitatively assess the risk of contamination by Pneumocystis depending on the degree of immunosuppression (ID) of the exposed rat hosts, we developed an animal model, where rats went through different doses of dexamethasone. Then, natural and aerial transmission of Pneumocystis carinii occurred during cohousing of the rats undergoing gradual ID levels (receivers) with nude rats developing pneumocystosis (seeders). Following contact between receiver and seeder rats, the P. carinii burden of receiver rats was determined by toluidine blue ortho staining and by qPCR targeting the dhfr monocopy gene of this fungus. In this rat model, the level of circulating CD4(+) and CD8(+) T lymphocytes remained significantly stable and different for each dose of dexamethasone tested, thus reaching the goal of a new stable and gradual ID rat model. In addition, an inverse relationship between the P. carinii burden and the level of circulating CD4(+) or CD8(+) T lymphocytes was evidenced. This rat model may be used to study other opportunistic pathogens or even co-infections in a context of gradual ID.

SYTO-13, a Viability Marker as a New Tool to Monitor In Vitro Pharmacodynamic Parameters of Anti-Pneumocystis Drugs.

While Pneumocystis pneumonia (PcP) still impacts the AIDS patients, it has a growing importance in immunosuppressed HIV-negative patients. To determine the anti-Pneumocystis therapeutic efficacy of new compounds, animal and in vitro models have been developed. Indeed, well-designed mouse or rat experimental models of pneumocystosis can be used to describe the in vivo anti-Pneumocystis activity of new drugs. In vitro models, which enable the screening of a large panel of new molecules, have been developed using axenic cultures or co-culture with feeder cells; but no universally accepted standard method is currently available to evaluate anti-Pneumocystis molecules in vitro. Thus, we chose to explore the use of the SYTO-13 dye, as a new indicator of Pneumocystis viability. In the present work, we established the experimental conditions to define the in vitro pharmacodynamic parameters (EC50, Emax) of marketed compounds (trimethoprim/sulfamethoxazole, pentamidine, atovaquone) in order to specifically measure the intrinsic activity of these anti-P. carinii molecules using the SYTO-13 dye for the first time. Co-labelling the fungal organisms with anti-P. carinii specific antibodies enabled the measurement of viability of Pneumocystis organisms while excluding host debris from the analysis. Moreover, contrary to microscopic observation, large numbers of fungal cells can be analyzed by flow cytometry, thus increasing statistical significance and avoiding misreading during fastidious quantitation of stained organisms. In conclusion, the SYTO-13 dye allowed us to show a reproducible dose/effect relationship for the tested anti-Pneumocystis drugs.

Complementation of a manganese-dependent superoxide dismutase-deficient yeast strain with Pneumocystis carinii sod2 gene.

Manganese-dependent superoxide dismutase (MnSOD) is one of the key enzymes involved in the cellular defense against oxidative stress. Previously, the Pneumocystis carinii sod2 gene (Pcsod2) was isolated and characterized. Based on protein sequence comparison, Pcsod2 was suggested to encode a putative MnSOD protein likely to be targeted into the mitochondrion. In this work, the Pcsod2 was cloned and expressed as a recombinant protein in EG110 Saccharomyces cerevisiae strain lacking the MnSOD-coding gene (Scsod2) in order to investigate the function and subcellular localization of P. carinii MnSOD (PcMnSOD). The Pcsod2 gene was amplified by PCR and cloned into the pYES2.1/V5-His-TOPO(®) expression vector. The recombinant construct was then transformed into EG110 strain. Once its expression had been induced, PcMnSOD was able to complement the growth defect of EG110 yeast cells that had been exposed to the redox-cycling compound menadione. N-term sequencing of the PcMnSOD protein allowed identifying the cleavage site of a mitochondrial targeting peptide. Immune-colocalization of PcMnSOD and yeast CoxIV further confirmed the mitochondrial localization of the PcMnSOD. Heterologous expression of PcMnSOD in yeast indicates that Pcsod2 encodes an active MnSOD, targeted to the yeast mitochondrion that allows the yeast cells to grow in the presence of reactive oxygen species (ROS).

An improved single-round PCR leads to rapid and highly sensitive detection of Pneumocystis spp.

In order to standardize a polymerase chain reaction (PCR)-based method of Pneumocystis detection, we describe the development of an improved PCR method that targets the Pneumocystis mtLSUrRNA gene. Design of a new primer pair and PCR program with suitable parameters and optimization resulted in a simpler and faster single-round amplification assay. The sensitivity of the novel Pneumocystis genus-specific PCR proved comparable to the reference nested PCR. The improvement that this new PCR assay offers in the detection and epidemiological studies of Pneumocystis spp. infection in research laboratories is discussed.