Perspectives of deprived patients on diabetes self-management programmes delivered by the local primary care team: a qualitative study on facilitators and barriers for participation, in France.

BACKGROUND: Diabetes self-management education (DSME) is an effective intervention for patients with type 2 diabetes mellitus (T2DM); nevertheless, patient participation in this type of programme is low. Implementation of DSME programmes in primary care practices by the local multi-professional team is a potential strategy to improve access to DSME for T2DM patients. The aim of this study was to identify perceived facilitators and barriers by patients to participation in local DSME delivered by primary care professionals in France. METHOD: T2DM patients, informed and recruited during consulting with their usual care provider, who had attended a structured and validated DSME programme delivered by 13 primary care providers within a multi-professional primary care practice in a deprived area of 20,000 inhabitants, were invited to participate in this study. A qualitative study with semi-structured, in-depth interviews was conducted with study participants, between July 2017 and February 2018. A reflexive thematic analysis of the interviews was carried out. Coding schemes were developed to generate thematic trends in patient descriptions of facilitators and barriers to DSME participation. RESULTS: Nineteen interviews (mean length 31 min; [20-44 min]) were completed with T2DM patients. Four themes on facilitators for programme participation emerged from the data: geographical proximity of a DSME programme held in the local multi-professional primary care practice; effective promotion of the DSME programme by the local multi-professional team; pre-existing relationship between patients and their healthcare providers; and potential to establish new social interactions within the neighbourhood by participating in the programme. Three themes on barriers to attendance emerged: integrating the DSME programme into their own schedules; difficulties in expressing themselves in front of a group; and keeping the motivation for self-managing their T2DM. CONCLUSIONS: From the patient perspective, the programme geographical proximity and the pre-existing patient-healthcare provider relationship were important factors that contributed to participation. Healthcare providers should consider these factors to improve access to DSME programmes and diabetes self-management in deprived populations. Longitudinal studies should be performed to measure the impact of these programmes.

Serosurvey for Brucella spp. and Coxiella burnetii in animals on Caribbean islands.

To determine the prevalence of antibodies to Brucella melitensis, Brucella abortus and Coxiella burnetii in animals on Caribbean islands we obtained sera from convenience samples of cattle (C), sheep (S), goats (G) and cats (F) from Dominica (C, S, G), Grenada (C, S, G), Montserrat (C, S, G), Puerto Rico (C), Nevis (C, S, G), St Kitts (C, S, G, F) and St Lucia (C, G). The sera were tested for antibodies against the Brucella spp. using commercial ELISA kits. Some sera were also tested at 1/80 for antibodies to C. burnetii using an indirect fluorescent antibody test. Positive sera were also tested at 1/640. None of 599 cattle, 462 sheep or 434 goats were positive in the Brucella ELISAs. None of 230 cattle had antibodies against C. burnetii, but one of 299 sheep was positive at 1/80 (Dominica - 1/54, 2%, 95% CI (0%-5.6%)), as were two of 314 goats, at 1/80 (Grenada - 1/53, 2%, 95% CI (0%-7.5%)) and 1/640 (St Kitts - 1/18, 5.6%, 95% CI (0%-16.7%)), and one of 34 cats, at 1/80 (St Kitts - 1/34; 3%, 95% CI (0%-8.8%)). Our data suggests that there is a very low prevalence or absence of B. melitensis and B. abortus on Caribbean islands. Coxiella burnetii, however, is present but it appears to be present on only some islands and then only at low levels. Overall, there appears to be a low threat to human and animal health from these organisms in the Caribbean.

Efficacy of slow-release tags impregnated with aggregation-attachment pheromone and deltamethrin for control of Amblyomma variegatum on St. Kitts, West Indies.

BACKGROUND: Amblyomma variegatum is an important cause of morbidity, mortality and economic losses in Africa and the West Indies. Attempts to control and/or eradicate the tick from the Caribbean have largely been unsuccessful because of difficulties relating to the biology of the three-host tick and problems with applying acaricides on a regular basis to free-ranging domestic ruminants. While plastic collars impregnated with insecticides are widely and effectively used in companion animals to control external parasites there is little information on this technology in ruminants. METHODS: Over 21 months we tested the efficacy of slow-release plastic tags impregnated with deltamethrin (7%) and aggregation-attachment pheromones (DPITs) in controlling A. variegatum on free-ranging cattle on two farms on St. Kitts. The tags were replaced every three months or when found to be lost. RESULTS: On sentinel animals fitted with tags containing only aggregation-attachment pheromones there were an average of 23.1 ticks per semi-monthly visit although this number varied considerably, peaking in the dry season around May and being lowest in August to October during the wet season. Significantly fewer ticks (3.5 on average) were found on cattle with DPITs at each visit (P < 0.001). Although the DIPTs provided good control (92% on average), they did not significantly reduce A. variegatum in the environment with tick numbers on sentinels being higher in the second year of the study, despite up to 44% of animals being fitted with DPITs. The tags were economical, costing 0.2% of the 1% flumethrin pour-on treatment widely recommended for A. variegatum control in the Caribbean. The major problem encountered was that 38% of tail tags were lost before they were due for replacement every three months. CONCLUSIONS: Our study has shown that DPITs are cheap to produce, easy to place, only require handling of animals every three months, and are very effective in protecting cattle from A. variegatum. Before DPITs can be considered for eradication programs the problems needing to be addressed include loss of tail tags, particularly in thick vegetation, and the optimum number of animals that must be treated to reduce numbers of ticks in the environment.

Ehrlichiosis, babesiosis, anaplasmosis and hepatozoonosis in dogs from St. Kitts, West Indies.

BACKGROUND: Although tick-borne diseases are important causes of morbidity and mortality in dogs in tropical areas, there is little information on the agents causing these infections in the Caribbean. METHODOLOGY: We used PCRs to test blood from a cross-section of dogs on St Kitts for Ehrlichia (E.) canis, Babesia (B.) spp., Anaplasma (A.) spp. and Hepatozoon (H.) spp. Antibodies against E. canis and A. phagocytophilum/platys were detected using commercial immunochromatography tests. Records of the dogs were examined retrospectively to obtain clinical and laboratory data. PRINCIPAL FINDINGS: There was serological and/or PCR evidence of infections of dogs with E. canis (27%; 46/170), Babesia spp. (24%; 90/372) including B. canis vogeli (12%; 43/372) and B. gibsoni (10%; 36/372), A. platys (11%; 17/157) and H. canis (6%; 15/266). We could not identify the Babesia sp. detected in nine dogs. There was evidence of multiple infections with dual infections with E. canis and B. canis vogeli (8%; 14/179) or B. gibsoni (7%; 11/170) being the most common. There was agreement between immunochromatography and PCR test results for E. canis for 87% of dogs. Only 13% of exposed dogs had signs of a tick-borne disease and 38% had laboratory abnormalities. All 10 dogs presenting for a recheck after treatment of E. canis with doxycycline were apparently healthy although all remained seropositive and six still had laboratory abnormalities despite an average of two treatments with the most recent being around 12 months previously. Infections with Babesia spp. were also mainly subclinical with only 6% (4/67) showing clinical signs and 13% (9/67) having laboratory abnormalities. Similarly, animals with evidence of infections with A. platys and H. canis were largely apparently healthy with only occasional laboratory abnormalities. CONCLUSIONS: Dogs are commonly infected with tick-borne pathogens in the Caribbean with most having no clinical signs or laboratory abnormalities.

Ehrlichia ruminantium in Amblyomma variegatum and domestic ruminants in the Caribbean.

The highly sensitive nested pCS20 polymerase chain reaction assay for Ehrlichia ruminantium was negative on 506 Amblyomma variegatum from Caribbean islands where clinical heartwater has not been reported, mainly the United States Virgin Islands (18), Dominica (170), Montserrat (5), Nevis (34), St. Kitts (262), and St. Lucia (17). Positive results were obtained with positive controls (Crystal Springs strain) and A. variegatum from countries in Africa where infections are endemic, mainly Tanzania (1/37) and Burkino Faso (2/29). Positive major antigenic protein-1 enzyme-linked immunosorbent assays for E. ruminantium were obtained on convenience samples of sera from apparently healthy cattle, sheep, and goats on Dominica (0/95, 0%; 3/135, 2%; 2/57, 4%), Grenada (0/4, 0%; 1/98, 1%; 1/86, 1%), Montserrat (0/12, 0%; 0/28, 2%; 5/139, 4%), Nevis (0/45, 0%; 0/157, 0%; 0/90, 0%), Puerto Rico (0/422, 0%; 0, 0%), St. Kitts (3/86, 4%; 1/25, 0%; 0/26, 0%), and St. Lucia (0/184, 0%; 0/15, 0%; 0, 0%), respectively. The pCS20 polymerase chain reaction results indicate E. ruminantium is not present on islands where clinical heartwater does not occur. The occasional positive major antigenic protein-1B enzyme-linked immunosorbent assay results appear, then, to be false-positive reactions, and serology appears to be of limited use in testing for E. ruminantium in the Caribbean, as is the case in Africa.

Rickettsia africae in Amblyomma variegatum and domestic ruminants on eight Caribbean islands.

We used PCRs with omp A primers to determine if spotted fever group rickettsiae occurred in Amblyomma variegatum from 6 Caribbean islands. Positive amplicons were obtained from ticks from the U.S. Virgin Islands (9/18; 50%), Dominica (39/171; 30%), Montserrat (2/5; 40%), Nevis (17/34; 50%), St. Kitts (46/227; 20%), and St. Lucia (1/14; 7%). Sequences for a convenience sample of reaction products obtained from A. variegatum on St. Kitts (7), American Virgin Islands (4), Montserrat (2), and St. Lucia (1) were 100% homologous with that of Rickettsia africae , the agent of African tick-bite fever. To determine if transmission of R. africae occurred, we used Rickettsia rickettsii antigen in IFA tests and found positive titers (≥ 1/80) with sera from cattle, goats, and sheep from Dominica (24/95 [25%], 2/136 [2%], 0/58 [0%]), Nevis (12/45 [27%], 5/157 [3%], 0/90 [0%]), St. Kitts (2/43 [5%], 1/25 [4%), 1/35 [3%]), and St. Lucia (6/184 [3%] cattle), respectively. No seropositive animals were found in Grenada (0/4, 0/98/, 0/86), Montserrat (0/12, 0/26, 0/52), or Puerto Rico (0/80 cattle). Our study indicates that R. africae and African tick-bite fever are widespread in the Caribbean.

Feline immunodeficiency virus, feline leukemia virus and Bartonella species in stray cats on St Kitts, West Indies.

Stray cats trapped in various areas of Basseterre, the capital of St Kitts in the West Indies, were tested for infection with feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) using commercial kits. Of 99 (51 male and 48 female) cats trapped in 2006/7, 15% (12 males and three females) were positive for FIV while none were positive for FeLV. Of 72 (41 males and 31 females) cats trapped in 2009, 14% (nine males and one female) were positive for FIV while none were positive for FeLV. Polymerase chain reaction analysis revealed DNA of Bartonella species in whole blood collected from 60/95 (63%) cats trapped in 2006/7. Sequencing of the 16S-23S rRNA gene intergenic transcribed spacer (ITS) region of a convenience sample of nine amplicons and the 11 isolates made from 43 blood samples which were cultured using Bartonella alpha Proteobacteria (BAPGM) enrichment medium revealed B henselae (14) and B clarridgeiae (six).

Failure to demonstrate Babesia, Anaplasma or Ehrlichia in thrombocytopenic dogs from St Kitts.

BACKGROUND: Thrombocytopenia is common in dogs on St Kitts but there is no data on the possible etiological role played by infections with Anaplasma, Babesia and Ehrlichia, other than E. canis, which are known to occur in the Caribbean. METHODOLOGY: Blood from 13 thrombocytopenic but apparently healthy dogs seronegative (Snap 3Dx) for E. canis were tested by PCR for Ehrlichia, Anaplasma and Babesia. RESULTS: All PCRs were negative. CONCLUSIONS: The results confirm the high sensitivity of SNAP testing for E. canis and indicate Anaplasma and Babesia are not important causes of thrombocytopenia in dogs on St Kitts.

Invasion of the Arabidopsis genome by the tobacco retrotransposon Tnt1 is controlled by reversible transcriptional gene silencing.

Long terminal repeat (LTR) retrotransposons are generally silent in plant genomes. However, they often constitute a large proportion of repeated sequences in plants. This suggests that their silencing is set up after a certain copy number is reached and/or that it can be released in some circumstances. We introduced the tobacco (Nicotiana tabacum) LTR retrotransposon Tnt1 into Arabidopsis (Arabidopsis thaliana), thus mimicking the horizontal transfer of a retrotransposon into a new host species and allowing us to study the regulatory mechanisms controlling its amplification. Tnt1 is transcriptionally silenced in Arabidopsis in a copy number-dependent manner. This silencing is associated with 24-nucleotide short-interfering RNAs targeting the promoter localized in the LTR region and with the non-CG site methylation of these sequences. Consequently, the silencing of Tnt1 is not released in methyltransferase1 mutants, in contrast to decrease in DNA methylation1 or polymerase IVa mutants. Stable reversion of Tnt1 silencing is obtained when the number of Tnt1 elements is reduced to two by genetic segregation. Our results support a model in which Tnt1 silencing in Arabidopsis occurs via an RNA-directed DNA methylation process. We further show that silencing can be partially overcome by some stresses.

Successful gene tagging in lettuce using the Tnt1 retrotransposon from tobacco.

The tobacco (Nicotiana tabacum) element Tnt1 is one of the few identified active retrotransposons in plants. These elements possess unique properties that make them ideal genetic tools for gene tagging. Here, we demonstrate the feasibility of gene tagging using the retrotransposon Tnt1 in lettuce (Lactuca sativa), which is the largest genome tested for retrotransposon mutagenesis so far. Of 10 different transgenic bushes carrying a complete Tnt1 containing T-DNA, eight contained multiple transposed copies of Tnt1. The number of transposed copies of the element per plant was particularly high, the smallest number being 28. Tnt1 transposition in lettuce can be induced by a very simple in vitro culture protocol. Tnt1 insertions were stable in the progeny of the primary transformants and could be segregated genetically. Characterization of the sequences flanking some insertion sites revealed that Tnt1 often inserted into genes. The progeny of some primary transformants showed phenotypic alterations due to recessive mutations. One of these mutations was due to Tnt1 insertion in the gibberellin 3beta-hydroxylase gene. Taken together, these results indicate that Tnt1 is a powerful tool for insertion mutagenesis especially in plants with a large genome.

Efficient transposition of the Tnt1 tobacco retrotransposon in the model legume Medicago truncatula.

The tobacco element, Tnt1, is one of the few active retrotransposons in plants. Its transposition is activated during protoplast culture in tobacco and tissue culture in the heterologous host Arabidopsis thaliana. Here, we report its transposition in the R108 line of Medicago truncatula during the early steps of the in vitro transformation-regeneration process. Two hundred and twenty-five primary transformants containing Tnt1 were obtained. Among them, 11.2% contained only transposed copies of the element, indicating that Tnt1 transposed very early and efficiently during the in vitro transformation process, possibly even before the T-DNA integration. The average number of insertions per transgenic line was estimated to be about 15. These insertions were stable in the progeny and could be separated by segregation. Inspection of the sequences flanking the insertion sites revealed that Tnt1 had no insertion site specificity and often inserted in genes (one out of three insertions). Thus, our work demonstrates the functioning of an efficient transposable element in leguminous plants. These results indicate that Tnt1 can be used as a powerful tool for insertion mutagenesis in M. truncatula.