TICKS PARASITIZING THE SPUR-THIGHED TORTOISE (TESTUDO GRAECA) POPULATION OF TUNISIA.

From March to April 2017, a total of 147 free-ranging spur-thighed tortoises (Testudo graeca) was captured in different habitat types of northern Tunisia and examined for tick infestation. A total of 134 was infested, yielding an infestation prevalence of 91.2%. From these tortoises, 1,174 ticks were collected, a subsample (10%, n=120) of which was randomly selected and identified; the remaining ticks were stored at -80 C and examined for zoonotic pathogens. Only adult Hyalomma aegyptium were found among the subsample of ticks collected for spur-thighed tortoise. The prevalence of tick infestation did not vary significantly within the study area but differences in intensity were observed in relation to the size of tortoises and the vegetation coverage. Our results provide strong evidence that adult H. aegyptium is the predominant tick species found on wild spur-thighed tortoises in northern Tunisia, where the species is commonly traded illegally across the Mediterranean Basin. Considering the potential transmission of several zoonotic pathogens by H. aegyptium to humans, we highlight the need to develop a surveillance system to prevent the introduction and the spread of tick-borne pathogens in the Mediterranean Basin.

Bayesian phylogenetic analysis of the influenza-A virus genomes isolated in Tunisia, and determination of potential recombination events.

The influenza-A virus (IAV) causes seasonal epidemics and presents a pandemic risk with the possibility of genetic re-assortment, allowing the emergence of new strains. The evolution of IAVes is done most often by relatively frequent re-assortment between gene segments, but the hypothesis of their evolution by recombination between RNA segments has not been justified to this date. Here, we examine this hypothesis by Bayesian phylogenetic analysis, to test if recombination events have occurred between genomic RNA segments. Different IAV subtypes are observed in co-circulation in Tunisia, which increases the probability of occurrence of double infections. Mixed infections are a prerequisite for recombination between co-infecting of viral strains. The aim of this work, and since understanding the evolutionary dynamics of IAV is essential for controlling human and avian influenza, phylogenetic analyzes (Bayesian approach) have been carried out for IAV strains isolated in Tunisia, to study their co-evolutionary history, trends, and possible recombination models. A set of IAV nucleic sequences, isolated in Tunisia from 2009 to 2013 (n = 102) were used in this study. These genomic segments encode various influenza A proteins. These viral strains studied were isolated following the 2009 H1N1 pandemic. The analyzes identified two large distinct groups of viral sequences and different subgroups. Assuming a relaxed molecular clock model (uncorrelated exponential (uced)) in a Bayesian coalescence approach and a constant effective time demographic history model (Coalescent: constant size), the substitution rate was estimated at 1.356 × 10-3 substitutions/site/year for segment 4 (haemagglutinin HA gene). Consistent estimates of the age of the most recent Common Ancestor (MRCA) were obtained for the different subgroups, the MRCA ages of the two viral populations corresponding to segment 4 and segment 6 (neuraminidase gene NA) of the genome are estimated at 443.737 years and 501.159 years respectively. A detailed phylogenetic study of the HA gene was performed. The incongruous phylogenetic models deduced for the three genomic subgroups studied corresponding to this gene were indicative of recombination events between the different subpopulations. The detection of these relative signals indicating the presence of recombination events can be considered as proof that recombination seems to play a role, even a small one, in the evolution of (IAV). Reliable recombination sites have been located with statistical significance between H3, H1 and H9 subtypes. MRCA age estimates of recombinants phylogenetic clades indicate directional gene transfers from the H1 and H9 populations to the H3 population, and from H1 and H3 to the H9 population, and their co-divergences during the study period.

Modeling and comparative study of the spread of zoonotic visceral leishmaniasis from Northern to Central Tunisia.

During the last twenty years, zoonotic visceral leishmaniasis (ZVL) spread from the north characterized by humid, sub-humid, and semi-arid bio-climates to the arid areas located in Central Tunisia. In order to understand the eco-epidemiological factors involved in the distribution of ZVL, both a sero-epidemiological and an entomological investigation were performed in two governorates situated in two different bio-geographical areas: Bizerte in the north and Kairouan in the center. A mathematical model for transmission of ZVL was built, describing the disease dynamic in these areas. Among 108 sera collected from clinically healthy owned dogs used for guarding houses in the governorate of Bizerte and tested for anti-L. infantum antibodies by indirect immune-fluorescence antibody test (IFAT), 8.3% were positive. From a total of 191 serum samples collected from clinically healthy owned dogs used for guarding houses in the governorate of Kairouan and tested for anti-L. infantum antibodies by IFAT, 26.7% were sero-positive. The cumulative incidences of ZVL in humans in the governorates of Bizerte and Kairouan were 39.17/100,000 and 0.2/100,000 inhabitants, respectively. The infection rates of sand flies with Leishmania DNA in the governorate of Bizerte and Kairouan were 3.8% and 9.4%, respectively. It is important to point out that ZVL epidemic in humans is preceded or concomitant to a high prevalence of Leishmania infantum in dogs and in sand flies. The model showed also a non-linear relationship between dog infection and ZVL in humans. These results suggest an important force of infection (R0) in emerging foci such as the governorate of Kairouan leading to an increased incidence of ZCL in humans. A high prevalence of L. infantum in dogs will result in high prevalence in the vector and obviously high probability to infect human; that it is a key factor for triggering transmission to humans, and subsequently it is an important parameter in the control of ZVL transmission.

Risk factors for mortality of children with zoonotic visceral leishmaniasis in Central Tunisia.

BACKGROUND: Zoonotic visceral leishmaniasis (ZVL) caused by Leishmania infantum is endemic with an epidemiological profile of a paediatric disease in Tunisia. In the context of a high fatality rate, identifying risk factors for in-hospital mortality in children treated for ZVL is of major epidemiological importance. DESIGN: A retrospective (case-control) study included 230 immuno-competent children diagnosed and confirmed with primary ZVL in the paediatric department of the University Hospital of Kairouan between 2004 and 2014. Forty-seven per cent (47%) were children under 18 months of age, and with a male / female ratio of 1.01:1. RESULTS: The overall case-fatality was 6% (n = 14). The risk factors for in-hospital death identified by a multivariate analysis were: bleeding at admission (OR = 25.5, 95% CI: 2.26-287.4; p = 0.009), white cell count less than 4000/mm3 (OR = 5.66, 95% CI: 1.16-27.6; p = 0.032), cytolysis (OR = 28.13, 95% CI: 4.55-173.6; p < 0.001), and delay between onset of symptoms and admission ≥ 15 days (OR = 11, 95% CI: 1.68-72; p = 0.012). CONCLUSION: The results strongly suggest that paediatric patients admitted 15 days after onset of symptoms, with bleeding, white cell counts below 4,000/mm3, and cytolysis at admission should be considered severe cases and subsequently, they are at high risk of mortality. A better understanding of factors associated with death of children from ZVL may contribute to decrease mortality.

Protective effects of Zygophyllum album extract against deltamethrin-induced hyperglycemia and hepato-pancreatic disorders in rats.

The current study was designed to investigate the possible mechanism involved in hyperglycemia induced by chronic exposure to deltamethrin (DLM) in rat and to assess whether this damage is amenable to modulation by Zygophyllum album. DLM, a synthetic pyrethroid pesticide, was administrated at a dose of 4 mg/kg body mass, during 60 days. Compared with control, DLM showed a significant increase of blood glucose (p ≤ 0.01) and glycosylated hemoglobin levels (p ≤ 0.01) and a clear decrease (p ≤ 0.01) of insulin and total hemoglobin levels. In addition, hepatic glycogen content and the activity of hexokinase decreased (p ≤ 0.01), whereas the activities of glucose-6-phosphatase and glycogen phosphorylase were significantly increased (p ≤ 0.01). Moreover, pancreatic lipid peroxidation (TBARS level) was higher (p ≤ 0.01) and oxidative stress biomarkers (SOD, CAT, GPx, and GSH) were altered owing to DLM toxicity. However, Z. album, when combined with DLM, significantly ameliorated almost all the hepato-pancreatic disorders induced by DLM alone. Furthermore, Z. album supplement was found to be effective in preserving the normal histological appearance of hepatic and pancreatic tissue. In conclusion, this study suggested that, owing to its antioxidant effects, methanolic extract of Z. album (MEZAL) can potentially prevent the hyperglycemia observed in DLM-treated group.

Non-cytotoxic Thymus capitata extracts prevent Bovine herpesvirus-1 infection in cell cultures.

BACKGROUND: Bovine herpes virus type 1 (BHV-1) still causes great economic loss to the livestock industry and trade because there aren't any available drugs that proved to be fully effective against it. In this study, the cytotoxicity and the antiviral activities of the Thymus capitata extracts were evaluated for the development of new, non toxic and specific anti-herpesvirus drug. Aqueous extracts (AE), ethanolic extracts (EE) and essential oil (EO) of the aerial parts of Thymus capitata were analyzed to determine their chemical compositions by gas chromatography, and high performance liquid chromatography combined with mass spectrometry. Their cytotoxicity and antiviral activities against Bovine Herpesvirus type 1 (BHV-1) were evaluated by quantifying the reduction of the viral cytopathic effect using Madin-Darby Bovine Kidney cell line with colorimetric assay. T. capitata extracts were added at different stages of the viral infection to investigate and better quantify their potential inhibitory effects. RESULTS: Polyphenols and flavonoids were the major compounds found in T. capitata EO, EE and AE. The cytotoxic concentrations at 50% were 48.70, 189 and 289 µg ml(-1) for EO, EE and AE, respectively. The inhibitor concentrations at 50% for the EO, EE and AE, were 3.36, 47.80 and 164 µg ml(-1), respectively. The selectivity index anti-BHV-1 values were 14.49, 3.95 and 1.81 for EO, EE and AE, respectively. Thus, the EO extracts were the most efficient antiviral compounds. T. capitata extracts affect mainly the adsorption of BHV-1 virus to host cells. CONCLUSION: T. capitata extracts inhibit the viral replication by interfering with the early stages of viral adsorption and replication. Thus, T. capitata is a potential candidate for anti-herpesvirus treatment.

Chemical Composition, Antioxidant and Antimicrobial Activities of Thymus capitata Essential Oil with Its Preservative Effect against Listeria monocytogenes Inoculated in Minced Beef Meat.

The chemical composition, antioxidant and antimicrobial activities, and the preservative effect of Thymus capitata essential oil against Listeria monocytogenes inoculated in minced beef meat were evaluated. The essential oil extracted was chemically analyzed by gas chromatography-mass spectrometry. Nineteen components were identified, of which carvacrol represented (88.89%) of the oil. The antioxidant activity was assessed in vitro by using both the DPPH and the ABTS assays. The findings showed that the essential oil exhibited high antioxidant activity, which was comparable to the reference standards (BHT and ascorbic acid) with IC50 values of 44.16 and 0.463 µ g/mL determined by the free-radical scavenging DPPH and ABTS assays, respectively. Furthermore, the essential oil was evaluated for its antimicrobial activity using disc agar diffusion and microdilution methods. The results demonstrated that the zone of inhibition varied from moderate to strong (15-80 mm) and the minimum inhibition concentration values ranged from 0.32 to 20 mg/mL. In addition, essential oil evaluated in vivo against Listeria monocytogenes showed clear and strong inhibitory effect. The application of 0.25 or 1% (v/w) essential oil of T. capitata to minced beef significantly reduced the L. monocytogenes population when compared to those of control samples (P-value <0.01).

Assessing the effect of zooprophylaxis on zoonotic cutaneous leishmaniasis transmission: a system dynamics approach.

Capturing or diverting the disease carrying vector from humans can reduce the transmission of vector borne diseases such as leishmaniasis. The use of animals that act as dead-end hosts to relieve the vector (sandfly) bites on humans is called zooprophylaxis. However, as the number of blood meal providers especially domestic animals increases, the sandflies enhanced availability of blood meals will improve its number and survival, thereby countering the impact of diverting bites from humans. Thus, the transmission model exhibits the structure of a feedback loop characterizing complex dynamic systems. In order to rigorously assess the effect of zooprophylaxis, we propose a system dynamic model for zoonotic cutaneous leishmaniasis transmission with 3 blood-meal hosts: domestic animals, humans, and a reservoir (rodents). In this context, a simulation study of the proposed model with a follow-up period of 1000 days was performed. We explored how perturbations in the parameters characterizing the transmission, essentially the vector biting rates and the size of the domestic animal population, affect the zooprophylaxis outcome. The results show that the basic reproductive number R0 and the disease incidence in humans are decreasing function of the relative size of the domestic animal population. The speed of this decrease depends also on the vector biting rates of the different mammal species. The key factors influencing the magnitude of zooprophylaxis are: the sizes of the vector, rodent, and domestic animal populations, as well as, the biting rates which incorporate relative attraction and accessibility of the vectors to the mammalian populations.

Prognostic value of Bcl-2 and Bax tumor cell expression in patients with non muscle-invasive bladder cancer receiving bacillus Calmette-Guerin immunotherapy.

Apoptosis is the distinctive form of programmed cell death that complements cell proliferation in maintaining normal tissue homeostasis. The significance of constitutive apoptosis in the recurrence of Non Muscle Invasive Bladder Cancer has yet to be investigated. The aim of this study is to investigate the prognostic significance of Bax and Bcl-2 in terms of recurrence after BCG immunotherapy. Immunohistochemical analysis was performed on frozen biopsies to evaluate bcl-2 and Bax proteins expression in 28 cases of NMIBC. All patients with confirmed NMIBC were treated with intravesical BCG-immunotherapy. The follow up was performed for 26 months. The correlation between clinicopathological, immunohistochemical data and the response to BCG therapy was performed. Univariate analysis showed that, PT1 stage, High grade and Bax expression increased significantly the risk of recurrence (P = 0.015, P = 0.015 and P= 0.034 respectively). In addition, multivariate analysis selected the model involving stage, age, Bax and Bcl-2 expression as the best independent variables of recurrence. In conclusion, the expression of Bcl-2 and Bax in NMIBC could have a prognostic value in assessing the risk of recurrence after BCG immunotherapy. These findings require further investigations on larger cohort in order to ascertain new molecular markers of the response to BCG immunotherapy.

Colonization of Phlebotomus papatasi changes the effect of pre-immunization with saliva from lack of protection towards protection against experimental challenge with Leishmania major and saliva.

BACKGROUND: Sand fly saliva has been postulated as a potential vaccine or as a vaccine component within multi component vaccine against leishmaniasis. It is important to note that these studies were performed using long-term colonized Phlebotomus papatasi. The effect of sand flies colonization on the outcome of Leishmania infection is reported. RESULTS: While pre-immunization of mice with salivary gland homogenate (SGH) of long-term colonized (F5 and beyond) female Phlebotomus papatasi induced protection against Leishmania major co-inoculated with the same type of SGH, pre-immunization of mice with SGH of recently colonized (F2 and F3) female P. papatasi did not confer protection against L. major co-inoculated with the same type of SGH. Our data showed for the first time that a shift from lack of protection to protection occurs at the fourth generation (F4) during the colonization process of P. papatasi. CONCLUSION: For the development of a sand fly saliva-based vaccine, inferences based on long-term colonized populations of sand flies should be treated with caution as colonization of P. papatasi appears to modulate the outcome of L. major infection from lack of protection to protection.

Evaluation and modeling of synergy to pheromone and plant kairomone in American palm weevil.

BACKGROUND: Many behavioral responses to odors are synergistic, particularly in insects. In beetles, synergy often involves a pheromone and a plant odor, and pest management relies on them for the use of combined lures. To investigate olfactory synergy mechanisms, we need to distinguish synergistic effects from additive ones, when all components of the mixture are active. RESULTS: As versatile tools and procedures were not available, we developed a bioassay, and a mathematical model to evaluate synergy between aggregation pheromone (P) and host plant odors (kairomone: K) in the American palm weevil, a pest insect showing enhanced responses to P+K mixtures. Responses to synthetic P and natural K were obtained using a 4-arm olfactometer coupled to a controlled volatile delivery system. We showed that: (1) Response thresholds were ca. 10 and 100 pg/s respectively for P and K. (2) Both stimuli induced similar maximum response. (3) Increasing the dose decreased the response for P to the point of repellence and maintained a maximum response for K. (4) P and K were synergistic over a 100-fold range of doses with experimental responses to P+K mixtures greater than the ones predicted assuming additive effects. Responses close to maximum were associated with the mixture amounts below the response threshold for both P and K. CONCLUSION: These results confirm the role of olfactory synergy in optimizing active host-plant localization by phytophagous insects. Our evaluation procedure can be generalized to test synergistic or inhibitory integrated responses of various odor mixtures for various insects.

Characteristics and pathways of bioactive 4-desmethylsterols, triterpene alcohols and 4α-monomethylsterols, from developing Tunisian cultivars and wild peanut (Arachis hypogaea L.).

Seven 4-desmethylsterols, five triterpene alcohols and three 4α-monomethylsterols were identified by GC-MS during the development of wild peanut, which is Arbi (AraA), and cultivars peanut, which are Trabelsia (AraT) and Chounfakhi (AraC). Our results showed that the maximum level of 4-desmethylsterols (881.07 mg/100 g of oil) was reached at 12 days after flowering (DAF) date of peanut plant in AraA, as well as the highest level of triterpene alcohols (31.51 mg/100 g of oil) was reached at 23 DAF in AraA, whilst, the highest level of 4α-monomethylsterols (15.11 mg/100 g of oil) was reached at 41 DAF in AraC. Herein, the level of triterpene alcohols and 4α-monomethylsterols was overwhelmed by the amount of 4-desmethylsterols at each stage of peanut maturity. Differences were observed in each sterol contents among the studied cultivars and wild one especially in immature stage.

Comparison of the concentrations of long-chain alcohols (policosanol) in three Tunisian peanut varieties (Arachis hypogaea L.).

Policosanol (PC) is a mixture of high molecular weight aliphatic primary alcohols. Literature about the contents and compositions of PC derived from peanut varieties is scarce. Total PC composition and content in whole peanut grain samples from three varieties of peanut (two cultivars, AraC and AraT, and a wild one, AraA) were identified using a gas chromatograph system coupled with a mass spectrophotometer. The results show that, qualitatively, 21 components of peanut aliphatic alcohols were identified (C14-C30). Besides (C18=), the results exhibited a previously unreported mixture of PC compositions in the peanuts: the unsaturated PC (UPC), which are (C20=), (C21=), (C22=), and (C24=). The main components of total PC in Tunisian peanut kernels are docosanol (C22), (Z)-octadec-9-en-1-ol (C18=), hexadecanol (C16), and octadecanol (C18). Quantitatively, the total PC content of the whole peanut samples varied from 11.18 to 54.19 mg/100 g of oil and was higher than those of beeswax and whole sugar cane, which are sources of dietary supplements containing policosanol.

Evaluation of the contribution of renin angiotensin system polymorphisms to the risk of coronary artery disease among Tunisians.

Recent studies have identified genetic markers that may directly influence the risk of the coronary artery disease (CAD), in particular the renin angiotensin system genes. Since there are no existing data for the Tunisian population, we investigated the association between these polymorphisms (angiotensin-converting enzyme [ACE] insertion/deletion [Ins/Del]; the angiotensinogen T174M and M235T; and the angiotensin II type 1 receptor A1166C polymorphisms) and CAD in Tunisians. Study subjects comprised 341 cases and 316 age- and sex-matched healthy individuals. Clinical characteristics and other biochemical and environmental risk factors were collected for both. The distribution of the Ins/Del genotypes was significantly different between cases and controls (p = 0.049) with the genotype Ins/Ins identified as a risk, p = 0.02. Similarly, the distributions of the T174M and M235T genotypes were significantly different between cases and controls (p = 0.037 and 0.047, respectively) with 174 M/M and 235 T/T as the risky genotypes (p = 0.001 and 0.026, respectively). However, A1166C genotype frequencies were not significantly different between patients and controls. In conclusion, our results suggest that a significantly higher risk of CAD was associated with the Ins/Del, the M235T, and T174M polymorphisms; other environmental variables such as body mass index; and biochemical variables such as cholesterol.

Lack of protection of pre-immunization with saliva of long-term colonized Phlebotomus papatasi against experimental challenge with Leishmania major and saliva of wild-caught P. papatasi.

Immunity to saliva of Phlebotomus papatasi protects against Leishmania major infection as determined by co-inoculation of parasites with salivary gland homogenates (SGHs) of this vector. These results were obtained with long-term colonized female P. papatasi. We investigated the effect of pre-immunization with SGH of long-term colonized P. papatasi against L. major infection co-inoculated with SGH of wild-caught P. papatasi. Our results showed that pre-exposure to SGH of long-term, colonized P. papatasi do not confer protection against infection with L. major co-inoculated with SGH of wild-caught P. papatasi. These preliminary results strongly suggest that the effectiveness of a vector saliva-based vaccine derived from colonized sand fly populations may be affected by inconsistent immune response after natural exposure.

Gas chromatography-mass spectrometry screening for phytochemical 4-desmethylsterols accumulated during development of Tunisian peanut kernels (Arachis hypogaea L.).

4-Desmethylsterols, the main component of the phytosterol fraction, have been analyzed during the development of Tunisian peanut kernels ( Arachis hypogaea L.), Trabelsia (AraT) and Chounfakhi (AraC), which are monocultivar species, and Arbi (AraA), which is a wild species, by gas chromatography-mass spectrometry. Immature wild peanut (AraA) showed the highest contents of beta-sitosterol (554.8 mg/100 g of oil), campesterol (228.6 mg/100 g of oil), and Delta(5)-avenasterol (39.0 mg/100 g of oil) followed by peanut cultivar AraC with beta-sitosterol, campesterol, and Delta(5)-avenasterol averages of 267.7, 92.1, and 28.6 mg/100 g of oil, respectively, and similarly for AraT 309.1, 108.4, and 27.4 mg/100 g of oil, respectively, were found. These results suggest that, in immature stages, phytosterol contents can be important regulator factors for the functional quality of peanut oil for the agro-industry chain from plant to nutraceuticals.

Bioinformatic characterizations and prediction of K+ and Na+ ion channels effector toxins.

BACKGROUND: K+ and Na+ channel toxins constitute a large set of polypeptides, which interact with their ion channel targets. These polypeptides are classified in two different structural groups. Recently a new structural group called birtoxin-like appeared to contain both types of toxins has been described. We hypothesized that peptides of this group may contain two conserved structural motifs in K+ and/or Na+ channels scorpion toxins, allowing these birtoxin-like peptides to be active on K+ and/or Na+ channels. RESULTS: Four multilevel motifs, overrepresented and specific to each group of K+ and/or Na+ ion channel toxins have been identified, using GIBBS and MEME and based on a training dataset of 79 sequences judged as representative of K+ and Na+ toxins.Unexpectedly birtoxin-like peptides appeared to present a new structural motif distinct from those present in K+ and Na+ channels Toxins. This result, supported by previous experimental data, suggests that birtoxin-like peptides may exert their activity on different sites than those targeted by classic K+ or Na+ toxins.Searching, the nr database with these newly identified motifs using MAST, retrieved several sequences (116 with e-value < 1) from various scorpion species (test dataset). The filtering process left 30 new and highly likely ion channel effectors.Phylogenetic analysis was used to classify the newly found sequences. Alternatively, classification tree analysis, using CART algorithm adjusted with the training dataset, using the motifs and their 2D structure as explanatory variables, provided a model for prediction of the activity of the new sequences. CONCLUSION: The phylogenetic results were in perfect agreement with those obtained by the CART algorithm.Our results may be used as criteria for a new classification of scorpion toxins based on functional motifs.

Zooprophylaxis: impact of breeding rabbits around houses on reducing the indoor abundance of Phlebotomus papatasi.

Zooprophylaxis is the use of animals to deviate vectors from humans. The indoor abundance of Phlebotomus papatasi in houses with rabbit holes in the peridomestic areas are significantly lower than the indoor abundance in houses without rabbit holes in their peridomestic areas. Introduction of rabbits in artificial underground holes in peridomestic areas reduced significantly the indoor abundance of P. papatasi. Cleaning rabbit holes in peridomestic area by removing all rabbit feces induced a significant increase in the abundance of P. papatasi inside bedrooms. The ecologic niche made around houses in endemic areas by creating active rabbit holes is a major source of attractiveness of P. papatasi, and therefore it may deviate the vector from humans to rabbits. Although rabbit holes are breeding sites for P. papatasi, rabbits are not competent reservoirs for Leishmania major. Our overall findings strongly suggest that zooprophylaxis could be effective in reducing the indoor abundance of P. papatasi and subsequently may be used to control the transmission L. major in rural areas.

Comparison of pp65 antigenemia, quantitative PCR and DNA hybrid capture for detection of cytomegalovirus in transplant recipients and AIDS patients.

The cytomegalovirus (CMV) antigenemia assay has been used frequently for rapid diagnosis of CMV infection, and antigenemia threshold values are recommended for triggering preemptive therapy. Hybrid capture of CMV's DNA and quantitative polymerase chain reaction (qPCR) are increasingly being adopted for early detection of CMV. The performance of the antigenemia assay, qPCR in plasma and hybrid capture in leukocytes were compared in 110 immunocompromised patients (38 bone-marrow transplants, 50 renal transplants and 22 AIDS patients). The most sensitive test was hybrid capture for transplants, while antigenemia and the qPCR showed similar performance for patients with AIDS. QPCR and hybrid capture thresholds requiring antiviral therapy were calculated using a receiver-operating-characteristic curve for antigenemia values corresponding to 2 positive cells for bone-marrow transplants and to 10 positive cells for renal transplants and AIDS patients. These threshold values varied with the group of patients considered, with corresponding sensitivities higher than 86% and specificities higher than 76% for hybrid capture, and sensitivities higher than 61% and specificities higher than 75% for qPCR in plasma. Hybrid capture in leukocytes can substitute for antigenemia in the case of transplants, and qPCR in plasma can substitute for it in the case of AIDS patients.

Natural infection of Phlebotomus (Larroussius) langeroni (Diptera: Psychodidae) with Leishmania infantum in Tunisia.

Phlebotomine sand flies were captured from an active transmission focus of sporadic cutaneous leishmaniasis, caused by Leishmania infantum, in El Kef region, northern Tunisia. Both Phlebotomus perniciosus and P. langeroni were found. Phlebotomus langeroni females showed a statistically significant intradomiciliary dominance (P<0.01 for the 2003 and 2004 seasons) when compared to animal shelters. During the 2003 season, dissection of collected female specimens showed the presence of flagellates within the digestive tracts of two P. perniciosus among 1086 observed, but none in 232 P. langeroni. Amplification of kinetoplast minicircles of Leishmania parasites was applied to DNA samples extracted from 298 frozen females including 249 P. perniciosus, 36 P. langeroni, 5 P. longicuspis and 8 P. perfiliewi and revealed by radioactive probe hybridization. Two P. langeroni females showed a signal of the size expected for L. infantum (800bp) indicating infection with these parasites. However, this PCR-hybridization method failed to identify any positive P. perniciosus females in pools of specimens. These results show for the first time the natural infection of P. langeroni with L. infantum in Tunisia, and support the existence of different L. infantum transmission cycles in Tunisia, with a potential role for P. langeroni as a vector.