Detection of Chlamydial {DNA} from Mediterranean Loggerhead Sea Turtles in Southern Italy

Chlamydiae are obligate intracellular bacteria that include pathogens of human and veterinary importance. Several reptiles were reported to host chlamydial agents, but pathogenicity in these animals still needs clarification. Given that only one report of chlamydiosis was described in sea turtles, and that chlamydiae might also be detected in hosts without clinical signs, the current study examined asymptomatic Mediterranean loggerhead sea turtles for the presence of chlamydial DNA. Twenty loggerhead sea turtles, rehabilitated at the Marine Turtle Research Centre (Portici, Italy), were examined collecting ocular-conjunctival, oropharyngeal and nasal swabs. Samples were processed through quantitative and conventional PCR analyses to identify Chlamydiales and Chlamydiaceae, with particular attention to C. pecorum, C. pneumoniae, C. psittaci, and C. trachomatis. Although it was not possible to determine the species of chlamydiae involved, the detection of chlamydial DNA from the collected samples suggests that these microorganisms might act as opportunistic pathogens, and underlines the role of sea turtles as potential carriers. This study highlights the presence of chlamydial agents in sea turtles, and encourages further research to fully characterize these microorganisms, in order to improve the management of the health and conservation of these endangered species, and prevent potential zoonotic implications

Lyme Borreliosis, Po River Valley, Italy

We aimed to determine the presence of Ixodes ricinus ticks in heavily populated areas of the Po River Valley after report of a Lyme disease case. Eighteen percent of ticks examined from 3 locations were positive for Lyme disease borreliae. Lyme disease was diagnosed for 3 workers at risk for tick bite

Phylogeography of Francisella tularensis subsp. holarctica, Europe

Francisella tularensis subsp. holarctica isolates from Austria, Germany, Hungary, Italy, and Romania were placed into an existing phylogeographic framework. Isolates from Italy were assigned to phylogenetic group B.FTNF002–00; the other isolates, to group B.13. Most F. tularensis subsp. holarctica isolates from Europe belong to these 2 geographically segregated groups

Demographic, clinical, and service-use characteristics related to the clinician’s recommendation to transition from child to adult mental health services

Purpose: The service configuration with distinct child and adolescent mental health services (CAMHS) and adult mental health services (AMHS) may be a barrier to continuity of care. Because of a lack of transition policy, CAMHS clinicians have to decide whether and when a young person should transition to AMHS. This study describes which characteristics are associated with the clinicians’ advice to continue treatment at AMHS. Methods: Demographic, family, clinical, treatment, and service-use characteristics of the MILESTONE cohort of 763 young people from 39 CAMHS in Europe were assessed using multi-informant and standardized assessment tools. Logistic mixed models were fitted to assess the relationship between these characteristics and clinicians’ transition recommendations. Results: Young people with higher clinician-rated severity of psychopathology scores, with self- and parent-reported need for ongoing treatment, with lower everyday functional skills and without self-reported psychotic experiences were more likely to be recommended to continue treatment. Among those who had been recommended to continue treatment, young people who used psychotropic medication, who had been in CAMHS for more than a year, and for whom appropriate AMHS were available were more likely to be recommended to continue treatment at AMHS. Young people whose parents indicated a need for ongoing treatment were more likely to be recommended to stay in CAMHS. Conclusion: Although the decision regarding continuity of treatment was mostly determined by a small set of clinical characteristics, the recommendation to continue treatment at AMHS was mostly affected by service-use related characteristics, such as the availability of appropriate services

Cohort profile : demographic and clinical characteristics of the MILESTONE longitudinal cohort of young people approaching the upper age limit of their child mental health care service in Europe

Purpose: The presence of distinct child and adolescent mental health services (CAMHS) and adult mental health services (AMHS) impacts continuity of mental health treatment for young people. However, we do not know the extent of discontinuity of care in Europe nor the effects of discontinuity on the mental health of young people. Current research is limited, as the majority of existing studies are retrospective, based on small samples or used non-standardised information from medical records. The MILESTONE prospective cohort study aims to examine associations between service use, mental health and other outcomes over 24 months, using information from self, parent and clinician reports. Participants: Seven hundred sixty-three young people from 39 CAMHS in 8 European countries, their parents and CAMHS clinicians who completed interviews and online questionnaires and were followed up for 2 years after reaching the upper age limit of the CAMHS they receive treatment at. Findings to date: This cohort profile describes the baseline characteristics of the MILESTONE cohort. The mental health of young people reaching the upper age limit of their CAMHS varied greatly in type and severity: 32.8% of young people reported clinical levels of self-reported problems and 18.6% were rated to be ‘markedly ill’, ‘severely ill’ or ‘among the most extremely ill’ by their clinician. Fifty-seven per cent of young people reported psychotropic medication use in the previous half year. Future plans: Analysis of longitudinal data from the MILESTONE cohort will be used to assess relationships between the demographic and clinical characteristics of young people reaching the upper age limit of their CAMHS and the type of care the young person uses over the next 2 years, such as whether the young person transitions to AMHS. At 2 years follow-up, the mental health outcomes of young people following different care pathways will be compared. Trial registration number: NCT03013595

Change in cognitive abilities over time during preschool age in low risk preterm children.

More information is needed on 'low-risk' preterm infants' neurological outcome so that they can be included in follow-up programs at least until school age. To examine the neuropsychological outcome in a group of low-risk low birth weight (LBW) children without neurological impairment followed from birth to 5 years of age. 26 intellectually normal children born preterm (30-34 weeks gestation) without major neurological disabilities and 23 control children born at term and matched for age, sex, and parental educational and occupational status. Subjects already evaluated at 3 years of age underwent assessment again at 5 years using as neuropsychological outcome measures a wide range of tests including perceptual and visual-motor function, language comprehension and expression, and attention skills. When tested at 5 years, children born preterm still obtained significantly lower mean scores than controls on visual motor integration test (57 vs 64, p=0.01), visual perception test (41 vs 43, p=0.002) and a trend toward a lower score in the picture vocabulary test (81 vs 85.5, p=0.07). The group of premature infants and controls improved their performance over time in the neuropsychological abilities investigated and, in some skills such as visual perception. Children born preterm took longer than those born at term to reach similar performance levels, 5 versus 3 years. Ex low-risk children born preterm achieve lower scores over time in visual-motor and perceptual ability scales and in some language tests than children born at term. Like high-risk premature infants even those at low risk deserve regular follow-up with long-term programs. Copyright © 2011 Elsevier Ltd. All rights reserved

High-resolution melt PCR analysis for rapid identification of <em>Chlamydia abortus</em> live vaccine strain 1B among <em>C. abortus</em> strains and field isolates

International audienceWe describe a novel high-resolution melt assay that clearly differentiates Chlamydia abortus live vaccine strain 1B from field C. abortus strains and field wild-type isolates based on previously described single nucleotide polymorphisms. This modern genotyping technique is inexpensive, easy to use, and less time-consuming than PCR-RFLP

Comparison among the Quantification of Bacterial Pathogens by qPCR, dPCR, and Cultural Methods

The demand for rapid methods for the quantification of pathogens is increasing. Among these methods, those based on nucleic acids amplification (quantitative PCRs) are the most widespread worldwide. Together with the qPCR, a new approach named digital PCR (dPCR), has rapidly gained importance. The aim of our study was to compare the results obtained using two different dPCR systems and one qPCR in the quantification of three different bacterial pathogens: Listeria monocytogenes, Francisella tularensis, and Mycobacterium avium subsp. paratuberculosis. For this purpose, three pre-existing qPCRs were used, while the same primers and probes, as well as PCR conditions, were transferred to two different dPCR systems: the QX200 (Bio-Rad) and the Quant Studio 3D (Applied Biosystems). The limits of detection and limits of quantification for all pathogens, and all PCR approaches applied, were determined using genomic pure DNAs. The quantification of unknown decimal suspensions of the three bacteria obtained by the three different PCR approaches was compared through the Linear Regression and Bland and Altman analyses. Our results suggest that, both dPCRs are able to quantify the same amount of bacteria, while the comparison among dPCRs and qPCRs, showed both over and under-estimation of the bacteria present in the unknown suspensions. Our results showed qPCR over-estimated the amount of M. avium subsp. paratuberculosis and F. tularensis cells. On the contrary, qPCR, compared to QX200 dPCR, under-estimated the amount of L. monocytogenes cells. However, the maximum difference among PCRs approaches was &lt;0.5 Log10, while cultural methods underestimated the number of bacteria by one to two Log10 for Francisella tularensis and Mycobacterium avium subsp. paratuberculosis. On the other hand, cultural and PCRs methods quantified the same amount of bacteria for L. monocytogenes, suggesting for this last pathogen, PCRs approaches can be considered as a valid alternative to the cultural ones

No evidence of transovarial transmission of Francisella tularensis by tick vectors Dermacentor reticulatus and Ixodes ricinus

Due to its easy dissemination, multiple routes of infection, high environmental contamination and morbidity and mortality rates, Francisella tularensis is considered a potential bioterrorism threat and classified as a category A select agent by the CDC. Tick bites are among the most prevalent modes of transmission, and ticks have been indicated as a possible reservoir, although their reservoir competence has yet to be defined. Tick-borne transmission of F. tularensis has been recognized since 1923, and transstadial transmission has been demonstrated in several tick species. Studies on transovarial transmission, conducted between 1920 and 1960, when molecular techniques had not been developed nor F. tularensis subspecies had been defined yet, have produced conflicting results. The aim of this study was to evaluate the role of ticks as reservoirs for Francisella, assessing the transovarial transmission of fully virulent F. tularensis subsp. holarctica in experimentallyinfected females of Dermacentor reticulatus and Ixodes ricinus. The experimental design was performed in 6 replicates. A total of 150 D. reticulatus and 150 I. ricinus unfed questing adult female ticks were used. For each replicate, 2 guinea pigs were used. On Day -3, all animals were infected with 25 female ticks and 35 male ticks and on Day 0, the tick-infested guinea pigs were inoculated subcutaneously with 500 CFU of the bacterium suspended in 0.3 ml of sterile saline solution. After completion of the tick blood feeding, bacterial culture and real-time PCR confirmed the infection by F. tularensis subsp. holarctica in all animals. All ticks examined during and/or at the end of oviposition were positive by PCR and culture. PCR, culture, transmission electron microscopy and fluorescence in situ hybridization showed F. tularensis within tick oocytes. However, cultures and bioassays of eggs and larvae were negative; in addition, electron microscopy techniques revealed bacterial degeneration/death in the oocytes. These results suggest that bacterial death might occur in oocytes, preventing the transovarial transmission of Francisella. We can speculate that Francisella does not have a defined reservoir, but rather various biological niches (e.g. ticks, mosquitoes, rodents, lagomorphs, amoebae) that allow the bacterium to persist in the environment. Our results suggest that ticks are not competent for the vertical transmission of the bacterium and are congruent with this view. However, even in the absence of a transovarial transmission, ticks are able to maintain the infection in the environment during the inter-epizootic period and can be identified as long-term vectors of F. tularensis