Molecular characterisation of Rhipicephalus sanguineus sensu lato ticks from domestic dogs in Nigeria.

Rhipicephalus sanguineus is the most widely reported tick in the world. Molecular characterisation is important to verify its taxonomic status in the different parts of the world. In this study, we provide information on the molecular characterisation of R. sanguineus tick of dogs collected from Nigeria. Ticks were collected from 62 of 93 sampled dogs. The collected ticks were subjected to morphological identification with the aid of appropriate entomological keys. Deoxyribonucleic acid (DNA) was extracted from the most prevalent tick species (R. sanguineus) and was subjected to further molecular characterisation protocols. The partial mitochondrial 16S rRNA gene sequences (∼300 bp) were obtained from representative specimens. Data were statistically analysed using the chi-square (χ2 ) test. Phylogenetic analysis was performed including different lineages of R. sanguineus (sl) from Africa, Asia, Europe and America, and other species belonging to the R. sanguineus 'tropical lineage' (R. linnaei) as well as Rhipicephalus turanicus and Ixodes ricinus. Results of this study showed that R. sanguineus was the most abundant ticks of dogs with a prevalence of 61.8% (68/110; 95% CI = 52.5-70.54), followed by Amblyomma variegatum (20.0%) and Haemaphysalis leachi (18.2%). The molecular analysis shows that they are genetically different from the temperate strains but closely related to those from other West African countries. There is a need to establish the vector competence of this common Nigerian dog tick.

Bartonella vinsonii subsp. arupensis infection in animals of veterinary importance, ticks and biopsy samples.

Testing for vector-borne pathogens in livestock is largely reliant upon blood and tissue. The role of biopsy samples remains poorly explored for detecting tick-borne bacteria in animals. In a 2-year survey, animals of veterinary importance from farms throughout the northern part of Greece were routinely checked for the presence of biopsy samples. Where detected, either a portion or a biopsy was collected together with whole blood samples and any ticks at the site of the biopsy sample. Molecular testing was carried out by real-time PCR targeting the internal transcribed spacer gene of Bartonella species. A total of 68 samples (28 blood samples, 28 biopsy samples and 12 ticks (nine Rhipicephalus bursa and three Rhipicephalus turanicus)) were collected from goats (64 samples) and cattle (four samples). Eight (11.8%) of the 68 samples were positive for Bartonella species. Of the biopsy and whole blood samples, four (14.3%) of each type were positive for Bartonella species. None of the ticks tested positive for Bartonella species. All pairs of positive biopsy samples/whole blood samples originated from the same animals. Positive samples were identified as Bartonella vinsonii subsp. arupensis. Although many more samples from a much wider spectrum of animal species is required before concluding upon the merit of biopsy samples in the study of tick-borne diseases, the significance of our finding warrants further study, both for clinical consequences in small ruminants and for those humans who are farming infected animals.

Public attitudes towards community pharmacy attributes and preferences for methods for promotion of public health services.

OBJECTIVE: To identify attitudes towards pharmacy characteristics and promotional methods for selected pharmacy public health services (lifestyle advice and screening for cardiovascular risk factors) among different sectors of the general public. STUDY DESIGN: Cross-sectional survey, using a previously validated questionnaire. METHODS: Three survey methods were used, across 15 areas of England, to maximize diversity: face-to-face; telephone; and self-completion of paper questionnaires. Responses to closed questions regarding characteristics and promotion were quantified and differences among sub-groups explored by univariate and multivariate analysis. RESULTS: In total, 2661 responses were available for analysis: 2047 face-to-face; 301 telephone; and 313 paper. There were strong preferences for a pharmacy near to home or doctor's surgery and for long opening hours, particularly among employed people and non-whites. Fifty percent preferred not to use a pharmacy in a supermarket, particularly older people, the retired, those of lower education and frequent pharmacy users. Personal recommendation by health professionals or family/friends was reported as most likely to encourage uptake of pharmacy public health services, with older people and males being less likely and frequent pharmacy users more likely to perceive any promotional method as influential. Posters/leaflets were preferred over mass-media methods, with fewer than 30% perceiving the latter as potentially influential. CONCLUSION: Pharmacists, pharmacy companies and service commissioners should use promotional methods favoured by potential users of pharmacy public health services and be aware of differences in attitudes when trying to reach specific population sub-groups. For personal recommendation to be successful, good inter-professional working and a pro-active approach to existing customers are needed.

The polymicrobial nature of biofilm infection.

The model of biofilm infection was first proposed over a decade ago. Recent scientific advances have added much to our understanding of biofilms, usually polymicrobial communities, which are commonly associated with chronic infection. Metagenomics has demonstrated that bacteria pursuing a biofilm strategy possess many mechanisms for encouraging diversity. By including multiple bacterial and/or fungal species in a single community, biofilms obtain numerous advantages, such as passive resistance, metabolic cooperation, byproduct influence, quorum sensing systems, an enlarged gene pool with more efficient DNA sharing, and many other synergies, which give them a competitive advantage. Routine clinical cultures are ill-suited for evaluating polymicrobial infections. DNA methods utilizing PCR methods, PCR/mass spectroscopy and sequencing have demonstrated their ability to identify microorganisms and quantitate their contribution to biofilms in clinical infections. A more robust model of biofilm infection along with more accurate diagnosis is rapidly translating into improved clinical outcomes.

Borrelia hispanica in Ornithodoros erraticus, Portugal.

Tick-borne relapsing fever (TBRF) is a spirochetal infection caused by the genus Borrelia. The disease is distributed in the Old and New World with many different species reported. In Europe, TBRF is caused by B. hispanica transmitted to man by Ornithodoros erraticus, a soft tick usually found in old premises to shelter pig herds. In Portugal, the first human case of TBRF was reported in 1942 but since the beginning of the 1960s, the disease has rarely been described and seems to either have disappeared or have been undiagnosed. Therefore, in 2009 a survey was undertaken to evaluate the presence of the tick in this type of premises and to evaluate its role as a reservoir of Borrelia. The work was carried out where the ticks were previously reported in the Alentejo and Algarve regions. Of 63 pigpens surveyed, O. erraticus was collected from 19% (n = 12) of these pigpens using CO(2) traps. To evaluate potential Borrelia hosts, both pigs (n = 25) and small rodents (n = 10) inhabiting these pigpens were surveyed for Borrelia presence, by whole blood PCR and/or tissue culture, respectively. All results for pigs and rodents were negative for the presence of B. hispanica. PCR assays targeting the 16S rRNA gene and intergenic spacer region of Borrelia were used. Sequence analysis of the positive samples confirmed the presence of B. hispanica in 2.2% (n = 5) of ticks from a pigpen in Alentejo. These results confirm natural, but albeit low, persistence of this agent in Portugal.

Does RecA have a role in Borrelia recurrentis?

Genomic sequencing of two relapsing fever spirochaetes showed truncation of recA in Borrelia recurrentis, but not in Borrelia duttonii. RecA has an important role among bacteria; we investigated whether this characteristic was representative of B. recurrentis, or an artefact following in vitro cultivation. We sequenced recA directly from samples of patient with louse-borne relapsing fever (B. recurrentis) or tick-borne relapsing fever (B. duttonii). We confirmed the premature stop codon in seven louse-borne relapsing fever samples, and its absence from three tick-borne relapsing fever samples. Furthermore, specific signature polymorphisms were found that could differentiate between these highly similar spirochaetes.

Characterizing the contribution of cardiac and hepatic uptake in dedicated breast SPECT using tilted trajectories.

A small field of view, high resolution gamma camera has been integrated into a dedicated breast, single photon emission computed tomography (SPECT) device. The detector can be flexibly positioned relative to the breast and image beyond the chest wall, allowing the system to capture direct views of the heart and liver. The incomplete sampling of these organs creates artifacts in reconstructed images, complicating lesion detection. To understand the limits imposed on a 3D acquisition trajectory, sequential tilted trajectories at increasing polar tilt are utilized to collect data of anthropomorphic phantoms filled with aqueous (99m)Tc in a clinically realistic concentration ratio. The counts collected per projection between different scans and the SNR, contrast and resolution (FWHM) of two hot lesions were compared. As expected, the counts per projection increased when the camera had direct views of the heart and liver, but remained relatively constant at other angles. The SNR, contrast and FWHM were more affected by the insufficient sampling of the data by the large polar angles than by the cardiac and hepatic activity. An upper bound on polar tilt for each azimuthal position reduces the artifacts in the reconstructed images. Such trajectories were implemented to show artifact-free reconstructed images.

Observer detection limits for a dedicated SPECT breast imaging system.

An observer-based contrast-detail study is performed in an effort to evaluate the limits of object detectability using a dedicated CZT-based breast SPECT imaging system under various imaging conditions. A custom geometric contrast-resolution phantom was developed that can be used for both positive ('hot') and negative contrasts ('cold'). The 3 cm long fillable tubes are arranged in six sectors having equal inner diameters ranging from 1 mm to 6 mm with plastic wall thicknesses of <0.25 mm, on a pitch of twice their inner diameters. Scans of the activity filled tubes using simple circular trajectories are obtained in a 215 mL uniform water filled cylinder, varying the rod:background concentration ratios from 10:1 to 1:10 simulating a large range of biological uptake ratios. The rod phantom is then placed inside a non-uniformly shaped 500 mL breast phantom and scans are again acquired using both simple and complex 3D trajectories for similarly varying contrasts. Summed slice and contiguous multi-slice images are evaluated by five independent readers, identifying the smallest distinguishable rod for each concentration and experimental setup. Linear and quadratic regression is used to compare the resulting contrast-detail curves. Results indicate that in a moderately low-noise 500 mL background, using the SPECT camera having 2.5 mm intrinsic pixels, the mean detectable rod was approximately 3.4 mm at a 10:1 ratio, degrading to approximately 5.2 mm with the 2.5:1 concentration ratio. The smallest object detail was observed using a 45 degrees tilted trajectory acquisition. The complex 3D projected sine wave acquisition, however, had the most consistent combined intra- and inter-observer results, making it potentially the best imaging approach for consistent results.

A calcitonin receptor (CALCR) single nucleotide polymorphism is associated with growth performance and bone integrity in response to dietary phosphorus deficiency.

Although concerns over the environmental impact of excess P in the excreta from pig production and governmental regulations have driven research toward reducing dietary supplementation of P to swine diets for over a decade, recent dramatic increases in feed costs have further motivated researchers to identify means to further reduce dietary P supplementation. We have demonstrated that genetic background impacts P utilization in young pigs and have identified genetic polymorphisms in several target genes related to mineral utilization. In this study, we examined the impact of a SNP in the calcitonin receptor gene (CALCR) on P utilization in growing pigs. In Exp. 1, 36 gilts representing the 3 genotypes identified by this CALCR SNP (11, 12, and 22) were fed a P-adequate (PA) or a marginally P-deficient (approximately 20% less available P; PD) diet for 14 wk. As expected, P deficiency reduced plasma P concentration, bone strength, and mineral content (P < 0.05). However, the dietary P deficiency was mild enough to not affect the growth performance of these pigs. A genotype x dietary P interaction (P < 0.05) was observed in measures of bone integrity and mineral content, with the greatest reduction in bone strength and mineral content due to dietary P deficiency being associated with the allele 1. In Exp. 2, 168 pigs from a control line and low residual feed intake (RFI) line were genotyped for the CALCR SNP and fed a PA diet. As expected, pigs from the low RFI line consumed less feed but also gained less BW when compared with the control line (P < 0.05). Although ADFI did not differ between genotypes, pigs having the 11 genotype gained less BW (P < 0.05) than pigs having the 12 or 22 genotypes. Pigs of the 11 and 12 genotypes had bones that tolerated greater load when compared with animals having the 22 genotype (P < 0.05). A similar trend was observed in bone modulus and ash % (P < 0.10). These data are supportive of the association of this CALCR SNP with bone integrity and its response to dietary P restriction. Although the allele 1 is associated with greater bone integrity and mineral content during adequate P nutrition, it is also associated with the greatest loss in bone integrity and mineral content in response to dietary P restriction. Understanding the underlying genetic mechanisms that regulate P utilization may lead to novel strategies to produce more environmentally friendly pigs.

Relapsing fever--a forgotten disease revealed.

Borrelial relapsing fever was once a major worldwide epidemic disease that made a significant impact on Livingstone during his epic travels through Africa and throughout Europe. Indeed, the term 'relapsing fever' was first used to describe clinical cases of this disease in Edinburgh. During the last century, we have witnessed the demise of the louse-borne infection, largely through improving standards of living resulting in a reduction in body lice, the vector for Borrelia recurrentis [louse-borne relapsing fever (LBRF)]. The tick-borne zoonotic form of the disease persists in endemic foci around the world [tick-borne relapsing fever (TBRF)]. Indeed, TBRF is reportedly the most common bacterial infection from Senegal and listed within the top ten causes of mortality in children under five in Tanzania. In Ethiopia, LBRF is again within the top ten causes of hospital admission, associated with significant morbidity and mortality. Despite these figures, many now regard relapsing fever as an unusual tropical disease. Certainly, recent cases have been imported following travel from endemic zones. More surprisingly, cases have been reported following family reunions in Colorado, USA. A further case was reported from the Mt Wilson observatory in Los Angeles, USA. In many regions, the infection is zoonotic with natural reservoirs in several vertebrate species. In West Africa, infection is again primarily zoonotic. Whether those species found predominantly in East Africa are zoonoses or are infections of humans alone is still debated, however, the life cycle may be determined by the feeding preferences of their arthropod vectors.

Shipboard testing of the efficacy of SeaKleen as a ballast water treatment to eliminate non-indigenous species aboard a working tanker in Pacific waters.

Trials were conducted aboard the tanker Seabulk Mariner to test a natural product, SeaKleen, as a biocide controlling non-indigenous populations of plankton and bacteria in ballast water. SeaKleen was dosed into matched ballast tanks at two different concentrations, 0.8 mg L(-1) active ingredient (a.i.) and 1.6 mg L(-1) a.i. during ballasting off the Oregon coast during a three-day passage to Prince William Sound, Alaska. Live organism counts from treated ballast water were compared with those from untreated (control tank) water collected from the same source location. Shipboard chemical analyses were made to verify dose and quantify chemical degradation and residuals following dilution. Results indicated that both SeaKleen doses resulted in complete zooplankton and phytoplankton mortality and that the higher dose (1.6 mg L(-1) a.i.) caused a two-log removal of culturable bacteria over a 92 h grow-out period. Spectrophotometry confirmed initial dosing to within 5% of nominal values. Shipboard bioassays were conducted using larval fish (Cyprinodon variegatus), brine shrimp (Artemia salina) and the bioluminescent dinoflagellate Pyrocystis lunula. Exposure of the test organisms to water drawn from treated ballast tanks 48 h after SeaKleen was added to the tanks resulted in 100% mortalities in Cyprinodon and Pyrocystis at both doses. Corresponding mortalities for Artemia larvae were 100% and 60% for high and low SeaKleen doses, respectively. Toxicity testing of treated water, subjected to varying dilutions, indicated that residual toxicity to even the most sensitive organisms would be eliminated once the discharge had dispersed beyond 100 feet from the vessel.

Myths, legends and realities of relapsing fever borreliosis.

Relapsing fever borreliosis is often shrouded in mystery. From its discovery, it has evaded fulfilment of Koch's postulates. It has resulted in epidemic waves of infection, although it is now mostly localized to particular endemic pockets of infection. Structurally, this spirochaete breaks many paradigms for conventional microorganisms, e.g. through its segmented genomic structure. Disclosure of host-microbial interactions is revealing a plethora of mechanisms, from antigenic variation to binding of various host-derived proteins. We dispel some of the myths and explore current understanding of this much neglected area through a series of reviews within this theme section.