Genetic diversity and haplotype analysis of Leishmania tropica identified in sand fly vectors of the genera Phlebotomus and Sergentomyia using next-generation sequencing technology.

Next-generation sequencing (NGS) was used to investigate the genetic diversity of Leishmania tropica in the sand fly vector, targeting the internal transcribed spacer 1 (ITS1) of the genus Leishmania. Bioinformatics analyses were conducted using Galaxy, MEGA version X, DnaSP ver. 6.12.03, and PopART 1.7 software for NGS analysis, phylogenetic tree, genetic diversity, and haplotype networking, respectively. A total of 307 engorged sand flies were trapped, with an overall Leishmania infection rate of 9.4 (29/307) and 6.8% by NGS and ITS1-PCR, respectively. Two Leishmania-infected sand fly genera were identified: Phlebotomus (10.2%, 26/254) and Sergentomyia (5.7% (3/53). The phylogenetic tree showed two clusters, cluster I included the four study sequences along with 25 GenBank-retrieved DNA sequences. Cluster II consisted of three sequences from Iran and Pakistan. The genetic diversity analysis for the 29 L. tropica sequences showed high haplotype (gene) diversity index (Hd) (0.62 ± 0.07) but low nucleotide diversity index (π) (0.04 ± 0.01). Tajima's D, a neutrality test, is more negative in cluster I (D = - 2.0) than in total population (D = - 1.83), but both are equally significant (P < 0.001), indicating that observed variation in cluster I and whole population is less frequent than expected. The median-joining haplotype network produced a total of 11 active haplotypes. In conclusion, L. tropica from sand flies in Palestine is monophyletic that assembled in one main phylogroup and one haplotype.

Tracking of SARS-CoV-2 Alpha variant (B.1.1.7) in Palestine.

As surges of the COVID-19 pandemic continue globally, including in Palestine, several new SARS-CoV-2 variants have been introduced. This expansion has impacted transmission, disease severity, virulence, diagnosis, therapy, and natural and vaccine-induced immunity. Here, 183 whole genome sequences (WGS) were analyzed, of which 129 were from Palestinian cases, 62 of which were collected in 11 Palestinian districts between October 2020 and April 2021 and sequenced completely. A dramatic shift from the wild type to the Alpha variant (B 1.1.7) was observed within a short period of time. Cluster mapping revealed statistically significant clades in two main Palestinian cities, Al-Khalil (Monte Carlo hypothesis test-Poisson model, P = 0.00000000012) and Nablus (Monte Carlo hypothesis test-Poisson model, P = 0.014 and 0.015). The phylogenetic tree showed three main clusters of SARS-CoV-2 with high bootstrap values (>90). However, population genetics analysis showed a genetically homogenous population supported by low Wright's F-statistic values (Fst <0.25), high gene flow (Nm > 3), and statistically insignificant Tajima's D values (Tajima's test, neutrality model prediction, P = 0.02). The Alpha variant, rapidly replaced the wild type, causing a major surge that peaked in April 2021, with an increased COVID-19 mortality rate, especially, in the Al-Khalil and Nablus districts. The source of introduction remains uncertain, despite the minimal genetic variation. The study substantiates the use of WGS for SARS-CoV-2 surveillance as an early warning system to track down new variants requiring effective control.

First Report of the HEV Seroprevalence and the Risk Factor Assessment in the West Bank, Palestine, during the Period of 2015-2017.

Hepatitis E virus is emerging viral hepatitis with hyperendemicity in many countries. Data on the burden of disease is not available in Palestine. This study aims to determine the seroprevalence and the risk factors of the HEV among the general population of the West Bank, Palestine. In this cross-sectional study, a total of 432 sera samples from 40 localities in the eleven districts of the West Bank and Jerusalem, Palestine, during the period of March 2015 to March 2017, were tested for HEV-IgG. A structured questionnaire was used to collect data of the participants' demographics and disease risk factors. The overall seroprevalence was 3.7%. Level of education was significantly inversely associated with HEV seropositivity (P=0.04). Purely spatial analysis did not detect any significant cluster related to the distribution of HEV-IgG cases; however, living in the southern West Bank is shown to be significantly associated with HEV. Age was also associated with HEV seropositivity. The young (<19 years) and adults (>40 years) had the highest prevalence, compared to those between 20 to 39 years old (P=0.12). Furthermore, males and those in contact with animals were associated with HEV seropositivity (P=0.1 and 0.3, respectively). In conclusion, the seroprevalence of HEV IgG in the West Bank, Palestine is low. Several well-investigated risk factors cannot be supported by our results due to the small number of the positive HEV-IgG samples. Finally, this study is useful for providing a first look into the seroepidemiology of HEV in Palestine.

Complete genome sequencing of SARS-CoV-2 strains: A pilot survey in Palestine reveals spike mutation H245N.

OBJECTIVES: SARS-CoV-2, severe respiratory syndrome coronavirus-2, is an RNA virus that emerged from China sweeping the globe in the form of a pandemic that became an international public health concern. This pilot study aimed to describe the genetic variation and molecular epidemiology of SARS-CoV-2 in Palestine in fall 2020. RESULTS: To achieve these aims, whole genome sequencing of SARS-CoV-2, phylogenetic analysis, haplotype networking and genetic diversity analysis were performed. These analyses revealed a unique spike mutation H245N that has never been reported before. The phylogenetic analysis depicted that three clusters existed in Palestinian SARS-CoV-2 genome sequences, in which cluster-I comprised the majority of clusters by 90%. Congruently, the haplotype network analysis depicted the same three clusters with a total of 39 haplotypes. The genetic diversity analysis showed that Cluster-I is highly diverse as confirmed by statistically significant mutation rate indices, Tajima's D and Fu-Li's-F tests (- 2.11 and 2.74, respectively), highest number of mutations (Eta = 120), highest number of haplotypes (h = 17), and highest average number of nucleotide differences between any two sequences (S = 118). The study confirmed the high genetic diversity among the Palestinian of SARS-CoV-2 which possessed high number of mutations including one which was reported for the first time.

Plasma Lipids and Lipoproteins in Sickle Cell Disease Patients in the Northern West Bank, Palestine.

BACKGROUND: Lipid metabolism may be altered in red cell genetic disorders. The erythrocyte and plasma lipids are defected which may increase the risk of cardiovascular disease. In the present study, we hypothesized a possible link between severity of anemia and altered lipid profile in SCD. METHODS: A total of 151 SCD patients, including 62 patients with sickle cell anemia (SS), 54 patients with sickle ß-thalassemia (ST), and 35 individuals with sickle cell trait (AS), were studied. The control group consisted of 160 healthy individuals. Total cholesterol (TC), triglyceride (TG), and high-density lipoprotein cholesterol (HDL-C) were enzymatically measured. RESULTS: Total cholesterol and LDL-C were significantly lower (P value < 0.001) in SS and ST patients compared to AS individuals and AA controls. However, LDL-C was significantly lower in AS individuals (both males and female) compared to AA controls (P value < 0.001). The HDL-C in SS and ST patients (both males and females) was significantly lower than that in AS individuals (P value < 0.001). In addition, the HDL-C was significantly higher in SS and ST males and AS (males and females) compared to AA controls (P value < 0.001). The HDL-C was also significantly higher in SS males (P value < 0.001) and females (P value < 0.05) compared to ST patients. The HDL-C was significantly higher in AS individuals (P value < 0.001) compared to AA controls. The triglycerides in SS males was significantly lower than that in ST patients (P value < 0.001), but there was no significant difference when compared to AS individuals and AA controls. In contrast, triglycerides in SS females were significantly lower than those in ST (P value < 0.05), AS (P value < 0.001), and AA controls (P value < 0.001). In males of ST patients, triglycerides were significantly higher than those observed in AS males and AA males (P value < 0.001). In contrast, females of ST patients have a significantly lower triglycerides compared to AS and AA females (P value < 0.001). CONCLUSIONS: In SCD, the plasma is affected in some way, especially the plasma cholesterol that was investigated in this study. Further prospective studies should examine the contribution of an altered lipid profile to the severity and clinical complications in SCD patients.

Genetic diversity, haplotype analysis, and risk factor assessment of hepatitis A virus isolates from the West Bank, Palestine during the period between 2014 and 2016.

BACKGROUND: Hepatitis A virus (HAV) infection is one of the major causes of acute viral hepatitis. HAV genotypes and its genetic diversity is rarely investigated in our region as well as worldwide. AIMS: The aims of the present study were to determine the HAV genotypes and its risk factors and to investigate the genetic diversity of the HAV isolates in the West Bank, Palestine. STUDY DESIGN: A cohort of 161 clinically and laboratory-confirmed HAV (IgM-positive) cases and 170 apparently healthy controls from all the districts of the West Bank, Palestine during the period of 2014 to 2016 were tested for HAV infection using IgM antibodies, RT-PCR and sequence analysis of the VP3/VP1 junction region of the HAV genome. Phylogenetic analysis, genetic diversity and haplotypes analysis were used to characterize the VP3/VP1 sequences. RESULTS: All the 34 sequences of the HAV were found to be of HAV-IB sub-genotype. The phylogenetic analysis showed four main clusters with cluster III exclusively consisting of 18 Palestinian isolates (18/23-78%), but with weak bootstrap values. A high haplotype diversity (Hd) and low nucleotide diversity (π) were observed. Cluster III showed high number of haplotypes (h = 8), but low haplotype (gene) diversity (Hd = 0.69). A total of 28 active haplotypes with some consisting of more than one sequence were observed using haplotype network analysis. The Palestinian haplotypes are characterized by closely related viral haplotypes with one SNV away from each other which ran parallel to cluster III in the phylogenetic tree. A smaller Palestinian haplotype (4 isolates) was three SNVs away from the major haplotype cluster (n = 10) and closer to others haplotypes from Iran, Spain, and South Africa. Young age, low level of parent's education, infrequent hand washing before meals, and drinking of un-treated water were considered the major HAV risk factors in the present study. CONCLUSION: Haplotype network analysis revealed haplotype variation among the HAV Palestinian sequences despite low genetic variation and nucleotide diversity. In addition, this study reconfirmed that age and parent's level of education as HAV risk factors, while hand washing and treating drinking water as protective factors.

Case Report: Autochthonous Case of Human Visceral Leishmaniasis in the West Bank, Palestine.

Human visceral leishmaniasis (HVL) is a parasitic disease infecting children in the Mediterranean region. Here, we portray a case of a 2-year-old child with an epidemiological description of the situation surrounding the case. The patient was suffering from recurrent fever, weakness, and abdominal discomfort associated with loss of appetite. Routine blood investigations showed pancytopenia, whereas examination revealed hepatomegaly. A diagnosis of HVL was made by demonstrating amastigotes in a Giemsa-stained smear from a bone marrow aspirate followed by genotyping by PCR and sequencing. In conclusion, early detection of VL infection followed by appropriate treatment protocols is essential to saving the patient.

Prevalence of selected intestinal protozoan infections in marginalized rural communities in Palestine.

BACKGROUND: Intestinal parasitic infections are common in rural areas with poor infrastructure and low socioeconomic status. The aim of this study was to estimate the prevalence of selected parasitic infections in marginalized rural areas in the northern part of the Palestinian West Bank Region, using conventional and PCR-based methods, and also to assess risk predictors of infection. METHODS: A cross-sectional study was conducted on 104 individuals from three rural villages in the Jordan Valley. Stool samples were collected and examined by a battery of tests that included microscopy of wet fecal samples in normal saline with iodine, concentration by ethyl acetate sedimentation and also by zinc sulfate floatation, a conventional PCR and a real-time PCR (qPCR). Risk factors were assessed that included demographic, socioeconomic, and behavioral characteristics. Data on method performance was analyzed by kappa-statistic, Cochrane's Q, and McNemar post hoc test. Mid-P exact test and odds ratio were used to discern association between outcome and risk predictors. RESULTS: The overall prevalence of intestinal parasitic infections was 48% (49/102). The predominant parasites were Giardia lamblia at 37% (37/102) and Hymenolepis nana at 9% (9/102). To concentrate cysts and eggs, sedimentation can be used as an alternative to floatation with a loss of 1% of positive cases. The methods employing PCRs proved crucial as it increased the detected infection rate of G. lamblia approximately three-fold from 13% by the conventional methods to 37% by the qPCR. Multiple infections were present in 13% (13/102) of the study group, which included double (10%) and triple (3%) infections. Regarding the genus Entamoeba, E. dispar and E. coli were detected at rates of 2 and 8%, respectively. While none of the individuals were infected with the pathogenic E. histolytica, E. nana (4%) was detected for the first time in the area. Age was a risk predictor for infection (OR = 2.61, CI 95% 1.05-6.45, P = 0.038). CONCLUSIONS: The increased prevalence of intestinal parasitic infections in children in marginalized rural areas in Palestine is worrying. The addition of PCR-based methods is important for the diagnosis of such infections as, with cautious interpretation, it increases proficiency and overcomes underestimation and misdiagnosis of cases. Control measures including education on personal hygiene and environmental sanitation, should be introduced to reduce the prevalence of the intestinal parasites and, thus, the infections they cause in this and other areas.

Genetic diversity of the enteroviruses detected from cerebrospinal fluid (CSF) samples of patients with suspected aseptic meningitis in northern West Bank, Palestine in 2017.

BACKGROUND: Human enterovirus genus showed a wide range of genetic diversity. OBJECTIVES: To investigate the genetic diversity of the enteroviruses isolated in 2017 in northern West Bank, Palestine. STUDY DESIGN: 249 CSF samples from aseptic meningitis cases were investigated for HEV using two RT-PCR protocols targeting the 5' NCR and the VP1 region of the HEV genome. The phylogenetic characterization of the sequenced VP1 region of Echovirus18 (E18) and Coxsackievirus B5 (CVB5) isolated in Palestine along with 27 E18 and 27 CVB5 sequences available from the Genbank were described. RESULTS: E18 and CVB5 account for 50% and 35% of the successfully HEV types, respectively. Phylogenetic tree of E18 and CVB5 showed three main clusters, with all Palestinian isolates uniquely clustering together with those from China and from different countries, respectively. Cluster I of E18, with 13 Palestinian and 6 Chinese isolates, showed the lowest haplotype-to-sequence ratio (0.6:1), haplotype diversity (Hd), nucleotide diversity (π), and number of segregating sites (S) compared to clusters II and III. Furthermore, cluster I showed negative Tajima's D and Fu-Li'sF tests with statistically significant departure from neutrality (P<0.01). In both E18 and CVB5 populations, high haplotype diversity, but low genetic diversity was evident. Inter-population pairwise genetic distance (Fst) and gene flow (Nm) showed that the Palestinian E18 and CVB5 clusters were highly differentiated from the other clusters. CONCLUSIONS: The study divulged close genetic relationship between Palestinian HEV strains as confirmed by population genetics and phylogenetic analyses.

Prevalence of Sero-Molecular Markers of Hepatitis C and B Viruses among Patients with ß-Thalassemia Major in Northern West Bank, Palestine.

BACKGROUND: HCV and HBV present a great challenge in the management of ß-thalassemia patients. OBJECTIVE: The present study aimed to determine the prevalence of both HBV and HCV in multitransfused-dependent ß-thalassemia patients in northern West Bank, Palestine, using sero-molecular markers. METHODS: Serum sample from 139 multitransfused ß-thalassemia patients were tested for HBV and HCV markers including HBsAg, anti-HBc, anti-HBs, HBV-DNA, and anti-HCV and HCV-RNA. Demographic data and selected clinical parameters were collected by means of a questionnaire and from the patients' medical files. RESULTS AND CONCLUSION: The mean (±SD) age of patients was 18.1 years (±10.6). The overall prevalence of the HCV was 10% (14/139), which is 50 times higher than the normal Palestinian population (0.2%). Of which, 3 were positive for anti-HCV alone, 7 positives for HCV-RNA alone, and 4 positives for both anti-HCV and PCR-RNA. On the other hand, low prevalence of HBV was detected at a level of 0.7% (1/139). Only one patient had HCV-HBV coinfection. Twenty-five patients (19%) were positive for anti-HBc, while 99 (71%) were immune with the anti-HBs level above 10 IU/mL. Anti-HBc was insignificantly high (P=0.07) in HCV-positive cases. In conclusion, the prevalence of HCV among ß-thalassemia patients is considered high compared to normal population. Determination of HCV prevalence should be based on the detection of both HCV-RNA and anti-HCV. On the contrary, HBV showed a low prevalence. A follow-up schedule and administration of booster dose of HBV vaccine is strongly recommended for ß-thalassemia patients whose anti-HBs level <10 IU/ml.

Frequency of Red Blood Cell Alloimmunization in Patients with Sickle Cell Disease in Palestine.

BACKGROUND: Transfusion of red blood cells (RBC) is an essential therapeutic tool in sickle cell disease (SCD). Repeated RBC transfusions can cause alloimmunization which causes difficulty in cross-matching and finding compatible blood for transfusions. This study aimed to investigate the frequency of RBC alloimmunization and related risk factors among Palestinian SCD patients. MATERIALS AND METHODS: A multicenter cross-sectional study on 116 previously transfused SCD patients from three centers in West Bank, Palestine. Demographic, medical data and history of transfusion were recorded. Blood samples were collected from transfused consenting SCD patients. Gel card method was used for antibody screening and identification. In all patients, autocontrol and direct antiglobulin (DAT) test were performed using polyspecific (anti-IgG + C3d) anti-human globulin (AHG) gel cards for the detection of autoantibodies. RESULTS: Of the SCD patients, 62 (53.4%) patients were HbSS and 54 (46.6%) patients were sickle ß-thalassemia (S/ß-thal). There were 53 (45.7%) females and 63 (54.3%) males. Mean age was 18.8 years (range 3-53 years). The frequency of RBC alloimmunization among SCD patients was 7.76%, with anti-K showing the highest frequency (33.3%) followed by anti-E (22.2%), anti-D (11.1%), anti-C (11.1%), and anti-c (11.1%). All reported IgG alloantibodies were directed against antigens in the Rh (66.7%) and Kell (33.3%) systems. Older ages of patients, increased number of blood units transfused, and splenectomy were the commonest risk factors for alloimmunization in our study. CONCLUSIONS: RBC alloimmunization rate among Palestinian SCD patients is low compared to neighboring countries and countries all over the world but still warrants more attention. Phenotyping of donors/recipients' RBC for Rh antigens and K1 (partial phenotype matching) before their first transfusion may reduce the incidence of alloimmunization.

The Prevalence of PCR-Confirmed Pertussis Cases in Palestine From Archived Nasopharyngeal Samples.

BACKGROUND: Pertussis caused by Bordetella pertussis is a vaccine-preventable disease causing whooping cough in humans of all ages. This study reports infection rate of pertussis in Palestine between the years 2004-2008 from archived nasopharyngeal samples collected from clinically- suspected cases. METHODS: A convenience archived DNA samples collected from 267 clinically-suspected pertussis cases were investigated for B. pertussis. Laboratory diagnosis was done by examining all DNA samples using polymerase chain reaction (PCR). RESULTS: Approximately 49% (130/267) were confirmed by PCR. A pertussis peak was shown to occur in 2008 with 77% (100/130) of PCR-confirmed cases isolated in that year. PCR-confirmed cases existed in all Palestinian districts with highest rate in Ramallah, Bethlehem, Jenin and Al-Khalil. Half of the PCR-confirmed cases (68/130) were less than 2 months old. The positivity rate among who had three doses of vaccine (at 2, 4 and 6 months) was 38%, and became 50% with the fourth dose at 12 months. CONCLUSION: The prevalence of pertussis was found to be significantly high among infants less than 2 months old. Active pertussis surveillance using rapid PCR assays is essential, as it is helpful in prompt diagnosis and treatment of patients with pertussis.

A comparison of the efficiency of three sampling methods for use in the molecular and conventional diagnosis of cutaneous leishmaniasis.

In human cutaneous leishmaniasis (CL), the success of positive diagnoses and species identifications depends, primarily, on how biopsies are taken and then processed and examined. The efficiency of three methods of taking skin biopsies from suspect cases of CL was compared using the classical methods of microscopy of stained smears, in vitro culture of tissue aspirate, and internal transcribed spacer region 1 (ITS1)-polymerase chain reaction in diagnosing positive cases and identifying the species of Leishmania causing them. From 1994-2014, biopsy samples from the skin lesions of 2232 CL-suspected patients were collected as unstained smears, as smears stained with Giemsa's stain and on filter paper, and compared in the diagnostic tests employed. Matched comparison based on testing biopsy samples from 100 patients, microscopy, in vitro culture and ITS1-PCR were also conducted to assess the most suitable combination of methods for diagnosing leishmaniases. In the 100-case-matched comparison, the three different types of sample proved to be equally good with no significant difference (P > 0.05). However, skin tissue imprints on filter paper revealed most cases of CL. The kappa statistic for measuring the degree of agreement among the three samples was 89%, which is considered good. Agreement was highest between imprints on filter paper and unstained smears, and lowest was for stained smears. In the overall comparison between the ITS1-PCR and conventional methods, the ITS1-PCR using samples from filter papers was the most sensitive method but the difference was insignificant (P = 0.32). The combination of microscopy together with ITS1-PCR on samples from filter papers increased the sensitivity significantly to 46%, compared to using the methods individually (P = 0.003-0.0008). On comparing the results of the tests done on the samples from the 2232 patients after applying ITS1-PCRs to their samples from filter papers, unstained smears, in vitro culture, microscopy, and stained smears showed, respectively, test sensitivities of 81, 69, 64, 57 and 48%. Of the tests and samples adjudicated, ITS1-PCRs run on skin tissue samples from filter papers proved best for the routine laboratory diagnosis of CL. Adding microscopy of stained smears to it, improved its diagnostic value significantly.

The clinical burden of human cystic echinococcosis in Palestine, 2010-2015.

BACKGROUND: Cystic echinococcosis (CE) is classified by the WHO as a neglected disease inflicting economic losses on the health systems of many countries worldwide. The aim of this case-series study was to investigate the burden of human CE in Palestine during the period between 2010 and 2015. METHODOLOGY/PRINCIPAL FINDINGS: Records of surgically confirmed CE patients from 13 public and private hospitals in the West Bank and Gaza Strip were reviewed. Patients' cysts were collected from surgical wards and formalin-fixed paraffin-embedded (FFPE) blocks were collected from histopathology departments. Molecular identification of CE species /genotypes was conducted by targeting a repeat DNA sequence (EgG1 Hae III) within Echinococcus nuclear genome and a fragment within the mitochondrial cytochrome c oxidase subunit 1, (CO1). Confirmation of CE species/genotypes was carried out using sequencing followed by BLAST analysis and the construction of maximum likelihood consensus dendrogram. CE cases were map-spotted and statistically significant foci identified by spatial analysis. A total of 353 CE patients were identified in 108 localities from the West Bank and Gaza Strip. The average surgical incidence in the West Bank was 2.1 per 100,000. Spot-mapping and purely spatial analysis showed 13 out of 16 Palestinian districts had cases of CE, of which 9 were in the West Bank and 4 in Gaza Strip. Al-Khalil and Bethlehem were statistically significant foci of CE in Palestine with a six-year average incidence of 4.2 and 3.7 per 100,000, respectively. CONCLUSIONS/SIGNIFICANCE: To the best of our knowledge, this is the first confirmation of human CE causative agent in Palestine. This study revealed that E. granulosus sensu stricto (s.s.) was the predominating species responsible for CE in humans with 11 samples identified as G1 genotype and 2 as G3 genotype. This study emphasizes the need for a stringent surveillance system and risk assessment studies in the rural areas of high incidence as a prerequisite for control measures.

Molecular detection and genotyping of enteroviruses from CSF samples of patients with suspected sepsis-like illness and/or aseptic meningitis from 2012 to 2015 in West Bank, Palestine.

BACKGROUND: Human enteroviruses (HEVs) are the most frequently reported cause of aseptic meningitis with or without CSF pleocytosis in childhood. Rapid detection and genotype of HEVs is essential to determine the causative agent and variant causing sepsis-like illness and/or aseptic meningitis. AIM: To investigate the molecular epidemiology of enteroviruses (EVs) among patients with sepsis-like illness and/or aseptic meningitis admitted to three major hospitals in West Bank, Palestine from 2012 to 2015. METHODS: During the study period, 356 CSF samples were collected from patients with sepsis-like illness and/or aseptic meningitis. Two RT-nested PCR assays targeting a partial part of 5'UTR for direct diagnosis and the VP1 region for genotyping by sequence analysis of the viral genome were used. RESULTS: HEV RNA was detected in 66 of 356 (18.5%) of CSF samples. Age distribution showed that 64% (42/66) were infants (<1 year), 18% were children between 1 and 5 years old, 12% were children between 5 and 10 years old, and 6% were more than 10 years old. Of the 66 EV cases, 12 were successfully genotyped. Five different EV genotypes were identified. All of them belonged to HEV-B species. The study showed that echovirus 6 genotype accounted for 42% of the sequenced cases. The HEV infections in the present study tended to show slight seasonal pattern with more cases occurring during spring and summer, yet still significant numbers were also reported in fall and winter seasons. CONCLUSION: HEV was isolated from a significant number of children with sepsis-like illness and/or aseptic meningitis. In addition, the molecular method utilized for direct diagnosis and genotyping of HEV from CSF revealed that more than one HEV type circulated in the West Bank, Palestine during the study period.

Incidence of Echinococcus granulosus in Domestic Dogs in Palestine as Revealed by Copro-PCR.

Hydatidosis or echinococcosisis considered a neglected zoonotic disease despite its high burden in the livestock industry and the high risk of infection by humans in endemic areas. In a cross-sectional study we estimated the copro-Incidence and also genotyped Echinococcus granulosus isolates from domestic dogs using polymerase chain reaction (PCR). Medical archives in nine major hospitals in Palestine were reviewed to determine incidence of E. granulosus infection detected in humans during surgery. Faecal samples were collected from 93 domestic dogs in three districts with the highest number of human cases: Al-Khalil (Hebron), Tubas and Jenin. Genomic DNA was extracted from dog faecal samples and amplified by PCR targeting the repeat DNA sequence (EgG1 Hae III) followed by sequencing of five positive samples. Genotyping was determined by sequencing and BLAST searching of mitochondrial cytochrome c oxidase subunit (CO1). The incidence of E. granulosus infection detected in humans at surgery was 1.2 per 100,000 in the West Bank and 1.0 per 100,000 in Gaza Strip. Seventeen of 93 domestic dogs (18%) were positive, based upon comparison with the Echinococcus DNA control. The five sequenced samples were confirmed to be E. granulosus. Successfully genotyped sample belonged to E.granulosus sensu stricto (formerly G1-G3 complex, sheep strain). For domestic dogs, age group (13-24 months) and sex were identified as two risk factors for contracting E. granulosus. The study identified the high incidence of E. granulosus sensu stricto in dogs in Palestine.

Persistence of Anti-HBs Among Palestinian Medical Students After 18 - 22 Years of Vaccination: A Cross-Sectional Study.

BACKGROUND: Hepatitis B infection is a global public health problem affecting various sectors in the society. Vaccination is the first line measure to prevent the disease. OBJECTIVES: To assess the persistence of anti-HBs marker among medical students after 18 - 22 of vaccination as an indicator for Hepatitis B virus vaccine efficacy. PATIENTS AND METHODS: In this study, 249 Palestinian medical students vaccinated at birth, 1, and 6 months of age using Engerix™-B starting from 1992 were studied. About 58% (144/249) of the students were Palestinians holding Israeli citizenship, while 42% (105/249) were Palestinians from the West Bank. Students were tested serologically for anti-HBs, as a marker for vaccine-induced immunity. RESULTS: Over 75% (188/248) of students had levels of anti-HBs greater than 10 mIU/mL indicating immunity and protection. Five cases had positive results for anti-HBc indicating exposure to HBV infection; however, none of these cases showed any sign of HBV-DNA indicating effective clearance of the virus by the vaccine. Around 57% of the study group had anti-HBs level of 100 - 1000 mIU/mL. No significant association was found between anti-HBs level and age, sex, locality and level of anti-HBc (P > 0.05). The students were aware of different aspects of hepatitis B infection regarding the virus, symptoms, prevention and mode of transmission. CONCLUSIONS: The Palestinian and Israeli official policies to give a booster dose for risk groups like medical students at anti-HBs level below 10 mIU/mL should continue to ensure absolute protection. The currently-used vaccine and its time program cleared virus from students believed to have been exposed to the virus during their lifetime.

Prevalence of occult HBV among hemodialysis patients in two districts in the northern part of the West Bank, Palestine.

Occult hepatitis B infection is the case with undetectable HBsAg, but positive for HBV DNA in liver tissue and/or serum. Occult hepatitis B infection among hemodialysis patients in Palestine has been understudied. In this study, 148 hemodialysis patients from 2 northern districts in Palestine, Jenin (89) and Tulkarem (59), were investigated for occult hepatitis B, HBV, HCV infections with related risk factors. ELISA and PCR were used for the detection of anti-HBc and viral DNA, respectively. The overall prevalence of occult hepatitis B infection among the study group was 12.5% (16/128). Occult hepatitis B infection is more prevalent among males with most cases (15/16) from Jenin District. About one-third (42/132) of the hemodialysis patients were anti-HBc positive. Approximately 27% of the hemodialysis patients were infected with HCV. Around 20% (28/140) were positive for HBV DNA, but only 8.2% (12/146) of the hemodialysis patients were positive for HBsAg. The comparison between hemodialysis patients with occult hepatitis B infection and those without occult hepatitis B infection for selected risk factors and parameters as liver Enzyme, age, sex, HCV infection, blood transfusion, kidney transplant, anti-HBc, and vaccination showed no statistical significance between both categories. Duration of hemodialysis significantly affected the rate of HCV infection. HCV is significantly higher in hemodialysis patients with both Diabetes mellitus and hypertension. The prevalence of occult hepatitis B infection among hemodialysis patients is high; requiring stringent control policies. HBsAg assay is insufficient test for accurate diagnosis of HBV infection among hemodialysis patients.

Echovirus 15 and autumn meningitis outbreak among children, Patras, Greece, 2005.

BACKGROUND: Enteroviruses are the most common cause of aseptic meningitis, presenting in epidemic or endemic form. OBJECTIVES: To determine the causative agent of an aseptic meningitis outbreak in autumn, 2005 in Patras, Greece. STUDY DESIGN: Cerebrospinal fluid (CSF) samples taken during May 2005-February 2006 from children admitted to the Children Hospital of Patras with signs of aseptic meningitis were tested for the presence of enteroviral RNA. Typing was performed by nucleotide analysis. RESULTS: Enteroviruses were detected in 11 (57.9%) of 19 tested CSF samples. In a 12-day period (27 October-7 November 2005) five aseptic meningitis cases were observed. Echovirus 15 was detected in all five cases, and differed from the prototype strain by 27.6%. Enteroviruses before and after this cluster of cases were of different serotypes (Echovirus 9, Echovirus 6). All patients with Echovirus 15 infection were male with a mean age of 7.7 years (2 months-13 years), all recovered successfully. CONCLUSIONS: This is the first report of a cluster of aseptic meningitis cases caused by Echovirus 15. The causative agent was a new variant of Echovirus 15.

Enterovirus meningitis in Greece from 2003-2005: diagnosis, CSF laboratory findings, and clinical manifestations.

Enteroviruses (EVs) are the most commonly identified cause of aseptic meningitis. Rapid detection and characterization of EV meningitis is essential in making decisions for patient management and treatment. A total of 52 cases of acute aseptic meningitis that occurred from March 2003 to April 2005 were investigated for EVs using viral culture and/or molecular methods directly in the cerebrospinal fluid (CSF). EVs were detected in 21 out of 52 (40.4%) patients using reverse transcription-PCR (RT-PCR) and/or tissue culture. EVs were isolated from six out of 37 (16.2%) cultured specimens, while 20 out of 52 (38.4%) specimens yielded positive results when 5'non-coding region (5'NCR) RT-PCR assay was used. One specimen that was culture-positive was RT-PCR-negative. Using the VP1-2A RT-PCR and sequence analysis, 14 of the 21 positive EVs were identified as: four strains of Coxsackie virus B5, five echovirus 11, two echovirus 9, one echovirus 5, one echovirus 14, and one Coxsackie virus A9. Fever, headache, vomiting, and stiff neck were the most pronounced symptoms. Pleocytosis with the predominance of lymphocytes and mild elevated protein levels characterized the CSF specimens. Coxsackie virus B5 and echovirus 11 were the predominant serotypes during the study period. Although there was seasonal enteroviral activity (April-November), cases also occurred in the cold months. The 5'NCR and VP1-2A RT-PCR with sequence analysis were found to be superior to conventional methods for direct diagnosis and the typing of EVs.