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Background Serotype 24F is one of the emerging pneumococcal serotypes after the introduction of pneumococcal conjugate vaccine (PCV). We aimed to identify lineages driving the increase of serotype 24F in France and place these findings into a global context. Methods Whole-genome sequencing was performed on a collection of serotype 24F pneumococci from asymptomatic colonisation (n=229) and invasive disease (n=190) isolates among individuals younger than 18 years in France, from 2003 to 2018. To provide a global context, we included an additional collection of 24F isolates in the Global Pneumococcal Sequencing (GPS) project database for analysis. A Global Pneumococcal Sequence Cluster (GPSC) and a clonal complex (CC) were assigned to each genome. Phylogenetic, evolutionary, and spatiotemporal analysis were conducted using the same 24F collection and supplemented with a global collection of genomes belonging to the lineage of interest from the GPS project database (n=25 590). Findings Serotype 24F was identified in numerous countries mainly due to the clonal spread of three lineages: GPSC10 (CC230), GPSC16 (CC156), and GPSC206 (CC7701). GPSC10 was the only multidrug-resistant lineage. GPSC10 drove the increase in 24F in France and had high invasive disease potential. The international dataset of GPSC10 (n=888) revealed that this lineage expressed 16 other serotypes, with only six included in 13-valent PCV (PCV13). All serotype 24F isolates were clustered in a single clade within the GPSC10 phylogeny and long-range transmissions were detected from Europe to other continents. Spatiotemporal analysis showed GPSC10-24F took 3–5 years to spread across France and a rapid change of serotype composition from PCV13 serotype 19A to 24F during the introduction of PCV13 was observed in neighbouring country Spain. Interpretation Our work reveals that GPSC10 alone is a challenge for serotype-based vaccine strategy. More systematic investigation to identify lineages like GPSC10 will better inform and improve next-generation preventive strategies against pneumococcal diseases. Funding Bill & Melinda Gates Foundation, Wellcome Sanger Institute, and the US Centers for Disease Control and Prevention.

The aim of this study was to comprehensively characterise S. aureus from the Caribbean Islands of Trinidad and Tobago, and Jamaica. A total of 101 S. aureus/argenteus isolates were collected in 2020, mainly from patients with skin and soft tissue infections. They were characterised by DNA microarray allowing the detection of ca. 170 target genes and assignment to clonal complexes (CC)s and strains. In addition, the in vitro production of Panton–Valentine leukocidin (PVL) was examined by an experimental lateral flow assay. Two isolates were identified as S. argenteus, CC2596. The remaining S. aureus isolates were assigned to 21 CCs. The PVL rate among methicillin-susceptible S. aureus (MSSA) isolates was high (38/101), and 37 of the 38 genotypically positive isolates also yielded positive lateral flow results. The isolate that did not produce PVL was genome-sequenced, and it was shown to have a frameshift mutation in agrC. The high rate of PVL genes can be attributed to the presence of a known local CC8–MSSA clone in Trinidad and Tobago (n = 12) and to CC152–MSSA (n = 15). In contrast to earlier surveys, the USA300 clone was not found, although one MSSA isolate carried the ACME element, probably being a mecA-deficient derivative of this strain. Ten isolates, all from Trinidad and Tobago, were identified as MRSA. The pandemic ST239–MRSA–III strain was still common (n = 7), but five isolates showed a composite SCCmec element not observed elsewhere. Three isolates were sequenced. That showed a group of genes (among others, speG, crzC, and ccrA/B-4) to be linked to its SCC element, as previously found in some CC5– and CC8–MRSA, as well as in S. epidermidis. The other three MRSA belonged to CC22, CC72, and CC88, indicating epidemiological connections to Africa and the Middle East.

Gram-negative bacterial infections are a global health problem. The production of beta-lactamase is still the most vital factor leading to beta-lactam resistance. In Trinidad and Tobago, extended spectrum beta-lactamase (ESBL) production has been detected and reported mainly in the isolates of Klebsiella pneumoniae and Escherichia coli and constitutes a public health emergency that causes high morbidity and mortality in some patients. In this literature review, the authors cover vast information on ESBL frequency and laboratory detection using both conventional and molecular methods from clinical data. The aim is to make the reader reflect on how the actual knowledge can be used for rapid detection and understanding of the spread of antimicrobial resistance problems stemming from ESBL production among common Gram-negative organisms in the health care system.

To assess colonization ofVaginal swabs were collected from 248 pregnant women attending antenatal clinics in northern Trinidad, and processed using standard microbiological laboratory tests to confirm GBS. Polymerase chain reaction detectedThe GBS carriage rate was 29% (72/248, 95% confidence interval 23.3-34.8), and carriage was significantly associated with variables including gestational diabetes (GBS colonization among pregnant women and resistance to commonly used antibiotics are high in Trinidad and Tobago. A population-based study is required to obtain accurate figures in order to improve maternal healthcare services.

Conventional commercially available rapid immuno-chromatographic tests (ICTs) or diagnostic kits were evaluated for their sensitivity, specificity, cost and turnaround time (TAT) results with Dengue IgM/IgG capture enzyme linked immunosorbent assays (ELISA) as the standard test, in blood samples from a cross-section of individuals with clinical features suggestive of dengue fever attending health care facilities in the country. Blood samples taken from over 100 consented participants were analyzed using the two rapid ICTs (SD Bioline Dengue Duo NS1/IgM/IgG and Panbio Dengue Duo Cassette) and compared with the Dengue IgM/IgG capture ELISAs. Standardized questionnaire was used to obtain bio and epidemiological data of the participants. The laboratory evaluation also assessed the TAT to complete the tests as well as the cost for each test method. The laboratory analysis on a given number (n=93) revealed that the SD Bioline was more sensitive (39.9%) than the Panbio (22.1%; p=0.005), and specificities for both were 100%. The SD Bioline includes an extra biomarker test with the same TAT and differs in cost by USD$ 1.14 as opposed to the Panbio. The ELISA has a cost of USD$ 8.07 and despite its longer TAT, it has the advantage of running more samples (1 vs 96) at a given time. While SD Bioline may be the better choice with a higher sensitivity, dengue ELISAs should also be favourably considered as an option for diagnostic purposes. In a resource strapped setting like the laboratories in Trinidad and Tobago, the ELISA should be preferred because its sensitivity and specificity were higher than the Panbio and SD Bioline kits. Besides, more samples were tested giving an effective TAT for amounts of samples completed despite a higher cost. Key words: Dengue fever, Panbio, SD Bioline, enzyme linked immunosorbent assays (ELISA), Trinidad and Tobago.

In this paper, the main objective was to raise chickens’ antibodies against three crucial public health microorganisms: the human immunodeficiency virus-1, Salmonella spp, and Staphylococcus aureus. Immunogens were prepared from the said microorganisms. Chickens were vaccinated either orally or intramuscularly. After a booster immunization, mostly eggs were collected and assess for the presence of specific antibodies. The most important results were the production of a large amount of anti-HIV antibodies in chicken’s eggs, and also the synthesis of anti-protein A antibodies with the ability to inhibit the growth of S. aureus in vitro and to serve as anti-anti-idiotypic antibodies with the capacity of neutralizing the original antigen. Enzyme-linked immune absorbent assays detected the presence of these antibodies as anti-Salmonella antibodies that were critical in reducing the bacterial load in the stomach and caeca compared with a control group. The vaccines were effective and safe, but more laboratory work, and economics have to be carried out to start a human trial.

Background: Invasive pneumococcal disease remains an important health priority owing to increasing disease incidence caused by pneumococci expressing non-vaccine serotypes. We previously defined 621 Global Pneumococcal Sequence Clusters (GPSCs) by analysing 20 027 pneumococcal isolates collected worldwide and from previously published genomic data. In this study, we aimed to investigate the pneumococcal lineages behind the predominant serotypes, the mechanism of serotype replacement in disease, as well as the major pneumococcal lineages contributing to invasive pneumococcal disease in the post-vaccine era and their antibiotic resistant traits. Methods: We whole-genome sequenced 3233 invasive pneumococcal disease isolates from laboratory-based surveillance programmes in Hong Kong (n=78), Israel (n=701), Malawi (n=226), South Africa (n=1351), The Gambia (n=203), and the USA (n=674). The genomes represented pneumococci from before and after pneumococcal conjugate vaccine (PCV) introductions and were from children younger than 3 years. We identified predominant serotypes by prevalence and their major contributing lineages in each country, and assessed any serotype replacement by comparing the incidence rate between the pre-PCV and PCV periods for Israel, South Africa, and the USA. We defined the status of a lineage as vaccine-type GPSC (≥50% 13-valent PCV [PCV13] serotypes) or non-vaccine-type GPSC (>50% non-PCV13 serotypes) on the basis of its initial serotype composition detected in the earliest vaccine period to measure their individual contribution toward serotype replacement in each country. Major pneumococcal lineages in the PCV period were identified by pooled incidence rate using a random effects model. Findings: The five most prevalent serotypes in the PCV13 period varied between countries, with only serotypes 5, 12F, 15B/C, 19A, 33F, and 35B/D common to two or more countries. The five most prevalent serotypes in the PCV13 period varied between countries, with only serotypes 5, 12F, 15B/C, 19A, 33F, and 35B/D common to two or more countries. These serotypes were associated with more than one lineage, except for serotype 5 (GPSC8). Serotype replacement was mainly mediated by expansion of non-vaccine serotypes within vaccine-type GPSCs and, to a lesser extent, by increases in non-vaccine-type GPSCs. A globally spreading lineage, GPSC3, expressing invasive serotypes 8 in South Africa and 33F in the USA and Israel, was the most common lineage causing non-vaccine serotype invasive pneumococcal disease in the PCV13 period. We observed that same prevalent non-vaccine serotypes could be associated with distinctive lineages in different countries, which exhibited dissimilar antibiotic resistance profiles. In non-vaccine serotype isolates, we detected significant increases in the prevalence of resistance to penicillin (52 [21%] of 249 vs 169 [29%] of 575, p=0·0016) and erythromycin (three [1%] of 249 vs 65 [11%] of 575, p=0·0031) in the PCV13 period compared with the pre-PCV period. Interpretation: Globally spreading lineages expressing invasive serotypes have an important role in serotype replacement, and emerging non-vaccine serotypes associated with different pneumococcal lineages in different countries might be explained by local antibiotic-selective pressures. Continued genomic surveillance of the dynamics of the pneumococcal population with increased geographical representation in the post-vaccine period will generate further knowledge for optimising future vaccine design. Funding: Bill & Melinda Gates Foundation, Wellcome Sanger Institute, and the US Centers for Disease Control. Fil: Lo, Stephanie W.. Wellcome Sanger Institute; Reino Unido Fil: Gladstone, Rebecca A.. Wellcome Sanger Institute; Reino Unido Fil: van Tonder, Andries J.. Wellcome Sanger Institute; Reino Unido Fil: Lees, John A.. University Of New York. School Of Medicine; Estados Unidos Fil: du Plessis, Mignon. National Institute For Communicable Diseases; Sudáfrica Fil: Benisty, Rachel. Ben Gurion University of the Negev; Israel Fil: Givon Lavi, Noga. Ben Gurion University of the Negev; Israel Fil: Hawkins, Paulina A.. University of Emory. Rollins School of Public Health; Estados Unidos Fil: Cornick, Jennifer E.. Malawi liverpool wellcome trust; Malaui Fil: Kwambana Adams, Brenda. University College London; Estados Unidos Fil: Law, Pierra Y.. University of Hong Kong; China Fil: Ho, Pak Leung. University of Hong Kong; China Fil: Antonio, Martin. Medical Research Council Unit The Gambia; Gambia Fil: Everett, Dean B.. University of Edinburgh; Reino Unido Fil: Dagan, Ron. Ben Gurion University of the Negev; Israel Fil: Von Gottberg, Anne. National Institute For Communicable Diseases; Sudáfrica Fil: Klugman, Keith P.. University of Emory. Rollins School of Public Health; Estados Unidos Fil: McGee, Lesley. Centers for Disease Control and Prevention; Estados Unidos Fil: Breiman, Robert F.. University of Emory. Rollins School of Public Health; Estados Unidos Fil: Bentley, Stephen D.. Wellcome Sanger Institute; Reino Unido Fil: Brooks, Abdullah W.. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Corso, Alejandra. The Global Pneumococcal Sequencing Consortium; Reino Unido. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina Fil: Davydov, Alexander. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Maguire, Alison. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Pollard, Andrew. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Kiran, Anmol. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Skoczynska, Anna. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Moiane, Benild. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Beall, Bernard. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Sigauque, Betuel. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Aanensen, David. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Lehmann, Deborah. The Global Pneumococcal Sequencing Consortium; Reino Unido Fil: Faccone, Diego Francisco. Dirección Nacional de Institutos de Investigación. Administración Nacional de Laboratorios e Institutos de Salud. Instituto Nacional de Enfermedades Infecciosas. Área de Antimicrobianos; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Objective: To determine the infective dose of Staphylococcus aureus (ATCC 29213) and Pseudomonas aeruginosa (ATCC 27853) in Sprague Dawley rats, when the inoculum is injected via the intraperitoneal route. Design and Methods: In interest of animal welfare and to reduce the number of animals used, we utilized the “up-down procedure” for dose determination of Pseudomonas aeruginosa and Staphylococcus aureus.Animals were grouped in fours, injected IP with inoculums of known concentration of a single organism and observed for 10 days. Clinical signs such as lethargy, increased respiration, porphyrin staining was recorded. Gross necropsy was performed ten (10) days post infection. Only one infective dose was done at a time and depending on the outcome, the size of inoculum was adjusted for the next step in the experimental infectious process. Results: Doses of inoculum was carefully titrated and the highest tolerable dose for each organism was determined. These doses allowed for survival of the animals and gave clinical signs, which mimicked the scenario in a human population. Symptoms of infection included lethargy, ruffled fur, porphyrin staining, dehydration and hunched back. At necropsy at 10 days post infection, common indicators of infection observed were ascites, abscesses on the intestinal wall, kidneys, liver and spleen. There were also fibrin tags, rounded livers, enlarged spleen and increased pericardial effusion was prominent in the S. aureusinfected group. Conclusions: The infective doses were determined based on clinical signs, survival and post mortem changes. The doses determined for Staphylococcus aureus (ATCC 29213) was 1.75 X 1010 cfu/ml and Pseudomonas aeruginosa (ATCC 27853) was 3.0 X 108cfu/ml. These doses would be useful in infective animal studies to determine the efficacy of antimicrobial agents.

To characterize extended-spectrum beta-lactamase (ESBL) and extra-intestinal pathogenic Escherichia coli (ExPEC) associated virulence genes in E. coli isolates from chickens and humans in Trinidad and Tobago. This cross sectional study was conducted over a three-month period. A total of 471 E. coli isolates; 160 from humans treated at a regional tertiary hospital and 311 from chicken caecal samples from “pluck shops” in Trinidad & Tobago were identified using both conventional and molecular microbiological methods. Phenotypic confirmation of ESBL producing E. coli isolates from humans was by Microscan system (Siemens, USA) while the double disk diffusion method was used for the chicken isolates. Polymerase chain reaction (PCR) analysis was used to determine the ESBL and ExPEC-associated virulence genes in representative human isolates and all chicken isolates. From the 311 chicken E. coli isolates, 49.2% (153/311) produced ESBL, while 56.3% (90/160) from humans were ESBL positive. All human and chicken ESBL isolates were 100% susceptible to carbapenems and aminoglycosides antimicrobials. PCR detected 21.1% blaCTX-M, 13.3% blaTEM and 7.8% blaSHV genes among E coli isolates from humans compared to 0.6% blaCTX-M and 48.6% blaTEM genes in chickens. PCR analysis revealed diverse virulence profiles among the isolates. There was a high occurrence rate of ExPEC-asso- ciated virulence genes in E. coli isolates from both humans and chickens. However, the CTX-M-1 genes were most predominant in humans while TEM occurred in chic- ken isolates. The diverse ESBL and virulence associated gene profiles encountered in E. coli isolates from humans and chickens on the surface depicts no similarity or relationships despite occurrence in both cohort groups. Therefore E. coli strains from chickens and humans require further investigation to determine their clonal relatedness or transmission in the country.

In this study we used spoligotyping and 15-loci MIRU-VNTRs for a finer characterization of Mycobacterium tuberculosis strains isolated from patients residing in Guyana (n = 74) and Suriname (n = 80). The mean age of the patients was 38.5 years (36.5 and 40.2 years for Guyana vs. Suriname), with a male-to-female sex-ratio of 2.25 for Guyana vs. 4.27 in Suriname. Spoligotyping and 15-loci MIRU-VNTRs led to a total of 41 and 65 different patterns respectively, with an overall clustering rate of 83.8% vs. 68.8%. Combined spoligotyping and VNTR analysis led to the detection of 18 clusters of 2–41 isolates, with an overall clustering of 67.5% and a recent “n − 1” transmission rate of 55.8%. Importantly, Guyana was characterized by a significantly higher percentage of clustered isolates than Suriname (79.7% vs. 56.3%; p = 0.0019). Three big spoligo/MIRU (SIT/MIT) clusters containing >10 isolates each were shared between the 2 countries, and concerned: T1 sublineage cluster 53/861 (n = 41, 37 in Guyana vs. 4 in Suriname); EAI6-BGD1 sublineage cluster 1340/860 (n = 16, 3 in Guyana vs. 13 in Suriname); and T1 sublineage cluster 131/146 (n = 11, 6 in Guyana vs. 5 in Suriname); as well as 2 smaller clusters of 2 and 3 isolates respectively. However, the relative phylogeographical specificities of strains in circulation as well as a lack of drug-resistance observed among strains from Suriname suggested that trans-border transmission of drug-resistant isolates occurred less frequently than thought. Tracing and interrupting transmission channels of a specific clone (SIT53/15-MIT861) should become a priority in Guyana, not only because it is by far most abundant but also because it accounts for almost half of the drug resistant isolates (n = 8/17, 47.1%) in our study, and clustered 5/12 (41.7%) MDR isolates.

Systemic lupus erythematosus (SLE) is a complex autoimmune disorder with a wide array of clinical manifestations including rash, photosensitivity, oral ulcers, arthritis, serositis, glomerulonephritis among others clinical findings. In this paper we globally summarized the most important epidemiological and clinical aspects to bear in mind, when the time comes to make the diagnosis of this rheumatic disorder and its management. Factor that are involved in the SLE pathogenesis and novel treatment options are mentioned.

Clostridium difficile is a Gram positive rod-shaped bacterium that produces two major toxins, A and B. The detection of the organism and its toxins has been widely carried out using specialized Enzyme-Linked Immunosorbent Assay (ELISA) kits; however these generally have been unsuccessful in identifying all Clostridium difficile positive samples. In this study, fifteen clinically symptomatic patients from three of the five major regional hospitals in Trinidad were investigated for Clostridium difficile infections. Stool samples were assessed by ELISA and cultured isolates were characterized using agar dilution antibiotic sensitivity assays, conventional Polymerase Chain Reaction (PCR), DNA sequencing and phylogenetic analysis of toxin A and B genes. All 15 patient stool samples and isolates were positive for toxigenic Clostridium difficile via ELISA and PCR respectively. All isolates were positive for the housekeeping tpi and Toxin B genes by PCR but only three of these were positive for the Toxin A gene. The Toxin B gene sequences showed 100% similarity levels among isolates while the Toxin A gene sequences showed 99% similarity among isolates. Phylogenetic analysis showed that the strains isolated in Trinidad most likely belonged to the same strain/group. Agar dilution sensitivity tests showed highest susceptibility to Pipercillin/Tazobactam and Meropenem (87%) and the highest resistance was seen with Cefotaxime in 93%. These results indicate that similar virulent strains of C. difficile are present in the Trinidad population and that pathogenic strains are more likely to be susceptible to Pipercillin/Tazobactam and Meropenem.

Significance USA300 is a hypervirulent, community-acquired, multidrug-resistant Staphylococcus aureus clone that started to spread in the United States around 17 years ago. Many studies detected it also in South America, Europe, and the Asia-Pacific region. In this study, we show that USA300 is also circulating in sub-Saharan Africa. Locating the temporal and spatial origin of clonal lineages is important with respect to epidemiology and molecular evolution of pathogens. We show that USA300 evolved from a less virulent and less resistant ancestor circulating in Central Europe around 160 years ago. Constant surveillance of pathogen transmission routes is vital to prevent and control potential outbreaks. Whole genome sequencing proved to be a useful tool for epidemiological surveillance., USA300 is a pandemic clonal lineage of hypervirulent, community-acquired, methicillin-resistant Staphylococcus aureus (CA-MRSA) with specific molecular characteristics. Despite its high clinical relevance, the evolutionary origin of USA300 remained unclear. We used comparative genomics of 224 temporal and spatial diverse S. aureus isolates of multilocus sequence type (ST) 8 to reconstruct the molecular evolution and global dissemination of ST8, including USA300. Analyses of core SNP diversity and accessory genome variations showed that the ancestor of all ST8 S. aureus most likely emerged in Central Europe in the mid-19th century. From here, ST8 was exported to North America in the early 20th century and progressively acquired the USA300 characteristics Panton–Valentine leukocidin (PVL), SCCmec IVa, the arginine catabolic mobile element (ACME), and a specific mutation in capsular polysaccharide gene cap5E. Although the PVL-encoding phage ϕSa2USA was introduced into the ST8 background only once, various SCCmec types were introduced to ST8 at different times and places. Starting from North America, USA300 spread globally, including Africa. African USA300 isolates have aberrant spa-types (t112, t121) and form a monophyletic group within the clade of North American USA300. Large parts of ST8 methicillin-susceptible S. aureus (MSSA) isolated in Africa represent a symplesiomorphic group of ST8 (i.e., a group representing the characteristics of the ancestor), which are rarely found in other world regions. Isolates previously discussed as USA300 ancestors, including USA500 and a “historic” CA-MRSA from Western Australia, were shown to be only distantly related to recent USA300 clones.

Molecular characteristics of vancomycin resistant enterococci isolates from Bermuda Island is currently unknown. This study was conducted to investigate phenotypic and genotypic characteristics of VRE isolates from Bermuda Island using the chromogenic agar, E-tests, polymerase chain reaction (PCR), pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Eighteen E. faecium isolates were completely analyzed and were all resistant to vancomycin, susceptible to linezolid and quinupristin/dalfopristin, positive for vanA and esp genes. The MLST analysis confirmed most isolates were of the sequence types linked to clonal complex 17 (CC17) that is widely associated with outbreaks in hospitals. Infection control measures, antibiotic stewardship, and surveillance activities will continue to be a priority in hospital on the Island.

Identification of the prevalence and spread of ESBL-mediated antibiotic resistance is essential especially in the hospital setting. It is for this reason, more and more studies are highlighting the importance of complementing phenotypic ESBL-detection techniques with molecular techniques in order to understand the basis and extent of this form of resistance among clinically evolved bacterial populations, especially those belonging to the Enterobacteriaceae family. However, in Trinidad and Tobago and other Caribbean countries, very little is known regarding ESBL detection rates and/or the prevalence of genes conferring ESBL resistance. Sixty-six Klebsiella pneumoniae isolates from clinical specimens phenotypically identified by the Microscan Walkaway-96 System as potential ESBL-producers were analysed in this study. Screening and confirmation of these isolates as ESBL producers was done by the Clinical and Laboratory Standards Institute (CLSI) approved methods. Polymerase chain reaction amplification of beta-lactamase genes bla TEM, bla SHV, bla CTX-M1, bla CTX-M2 and bla AmpC was performed to identify mechanisms of β-lactam resistance. ESBL-producing K. pneumoniae was confirmed in 78.8% (41/52) from isolates collected from a variety of sources during the period, April–July 2015. bla SHV (84.8%) and bla CTX-M (46.9%) were the predominant β-lactamase genes identified. A single K. pneumoniae isolate possessed a bla CTX-M group 2 beta-lactamase gene. RAPD analysis identified a number of epidemiologically related isolates. However, current isolates were unrelated to isolates from previous years. This study revealed that among K. pneumoniae isolates exhibiting extended spectrum β-lactam resistance, there was a high prevalence of bla SHV and bla CTX-M genes. This result highlights the need for a reliable epidemiological apparatus that involves the molecular characterisation of ESBL resistance.

Emergence of vancomycin-resistant Enterococci (VRE) that first appeared on the stage about three decades ago is now a major concern worldwide as it has globally reached every continent. Our aim was to simply undertake a multinational study to delineate the resistance and virulence genes of clinical isolates of VRE isolates from the Caribbean. We employed both conventional (standard microbiological methods including use of E-test strips, chromogenic agar) and molecular methods (polymerase chain reactions–PCR, pulsed-field gel electrophoresis–PFGE and multilocus sequence typing–MLST) to analyze and characterize 245 Enterococci species and 77 VRE isolates from twelve hospitals from eight countries in the Caribbean. The PCR confirmed and demonstrated the resistance and virulence genes (vanA and esp) among all confirmed VRE isolates. The PFGE delineated clonally related isolates from patients from the same country and other countries in the region. The main sequence types of the VRE isolates from the region included STs 412, 750, 203, 736 and 18, all from the common ancestor for clonal complex 17 (CC17). Despite this common ancestor and association of outbreaks of this lineage clones, there has been no reports of outbreaks of infection by VRE in several hospitals in the Caribbean.

Salmonella infection in bird species in Jamaica was studied. This revealed that very low prevalence of salmonellosis was found (0.32 %). Salmonella Yeerongpilly (newly reported in the country) was isolated from a bird collected at a bird aviary. This study showed that there was the presence of this Salmonella serovar in a Chinese owl (Columba livia domestica) in Jamaica. There were not published reports from Caribbean Islands of the presence of this serovar. Salmonella Yeerongpilly belongs to serogroup E1 and by molecular serotyping random amplification of polymorphic DNA (RAPD) fingerprinting belongs to A20, B17 and C21. This strain was isolated in Queensland Australia in the 1960s before the successful Salmonella eradication campaign. This study suggests that a larger investigation in pet birds as Salmonella carriers should be carried out in Jamaica. Mandatory screening or quarantine of birds entering the country should be institutionalized. Original Research Article British Microbiology Research Journal, 4(2): 167-172, 2014 168

A 56-year-old woman who vaccinated as a child with the Bacillus Calmette-Guerin (BCG), now tests positive to the tuberculin skin test (TST) but test negative to the Quantiferon Gold assay. She has no history of tuberculosis contact and is asymptomatic. This dilemma now is, should be treated for tuberculosis or not, based only on the TST results? To prevent these falsepositive results with TST and avoid treatment with isoniazid (INH) it may be helpful to use interferon-gamma release assay (IGRA) instead, which unlike the TB skin test is not affected by prior BCG vaccination. Keywords: Bacillus calmette-guerin, Tuberculosis, Tuberculin skin test, Purified protein derivative, Mantoux test, Quantiferon, Gold, Isoniazid

Panton-Valentine leukocidin (PVL) is a virulence factor of Staphylococcus aureus , which is associated with skin and soft-tissue infections and necrotizing pneumonia. To develop a rapid phenotypic assay, recombinant PVL F component was used to generate monoclonal antibodies by phage display. These antibodies were spotted on protein microarrays and screened using different lukF-PV preparations and detection antibodies. This led to the identification of the optimal antibody combination that was then used to establish a lateral flow assay. This test was used to detect PVL in S. aureus cultures. The detection limit of the assay with purified native and recombinant antigens was determined to be around 1 ng/ml. Overnight cultures from various solid and liquid media proved suitable for PVL detection. Six hundred strains and clinical isolates from patients from America, Europe, Australia, Africa, and the Middle East were tested. Isolates were genotyped in parallel by DNA microarray hybridization for confirmation of PVL status and assignment to clonal complexes. The sensitivity, specificity, and positive and negative predictive values of the assay in this trial were 99.7, 98.3, 98.4, and 99.7%, respectively. A total of 302 clinical isolates and reference strains were PVL positive and were assigned to 21 different clonal complexes. In summary, the lateral flow test allows rapid and economical detection of PVL in a routine bacteriology laboratory. As the test utilizes cultures from standard media and does not require sophisticated equipment, it can be easily integrated into a laboratory's workflow and might contribute to timely therapy of PVL-associated infections.

The Human Papilloma Virus (HPV) and Human Immunodeficiency Virus (HIV) are both sexually transmitted infections, which have impacted the prevalence of cervical dysplasia and cancer in women. Infections with one of these viruses can facilitate infection with the other. In Jamaica cervical cancer is seen in 27.5 per 100, 000 women making it the second leading cause of cancer death in this population only to breast cancer as a cause of death in women with cancer. Our study investigates the seroprevalence of anti-HIV antibodies in women with abnormal pap smears in Jamaica to determine the influence of HIV on cervical dysplasia. Only patients with positive confirmatory tests were classified as HIV positive. Enzyme-Linked Immunosorbent Assay (ELISA) was used for screening while the Western blot was used for confirmation. Sero-prevalence of anti-HIV antibodies in women with abnormal pap smears was 0.85%. The preliminary results of HIV

Aims: The aim of this study was to investigate the effectiveness of medication in the prevention of Salmonella in growing chickens. Material and Methods: All specimens were placed in an igloo with ice packs and transported to the laboratory for analysis. Bacteriological media obtained from Difco Laboratories Detroit MI U.S.A were used for the isolation and identification of Salmonella spp. Salmonella Serological typing was performed to determine the Salmonella serovar by standard procedures. Chickens were given a single dose of a wild-type multiresistant strain of Salmonella Typhimurium (resistant to many antibiotics including cloramphenicol and bactrim) by oral route (3x10 9 cfu/bird) as a challenge. Results: The effectiveness of some medications in the reduction of Salmonella infections in growing chicken revealed that none of the groups including the control group showed

Aim: This study was undertaken to determine the prevalence of Salmonella spp contamination in the Jamaican poultry industry and its environments. Materials and Methods: A total of 45 farms across 6 Jamaican parishes were selected for this study. A total of 6693 specimens from animals and the environment were investigated for the presence of Salmonella spp. All specimens were placed in an igloo with ice packs and transported to the laboratory for analysis. Bacteriological media obtained from Difco Laboratories Detroit MI U.S.A were used for the isolation and identification of Salmonella spp. Salmonella serological typing was performed to determine the Salmonella serovar by standard procedures. Results: This study revealed a low prevalence of Salmonella contamination/infection in both small and large entities in the poultry industry in Jamaica. The overall prevalence was 1 % (79 positive out of 6693 specimens). However, a higher prevalence of Salmonella was observed in the case of those operations which practiced “organic” poultry farming. It was shown that two Salmonella serovars including Augustenborg and Kentucky, identified Research Article British Microbiology Research Journal, 3(4): 461-469, 2013 462 during the study, are newly reported serovars in Jamaica. The sources of Salmonella infection varied from poultry itself to other species, such as rodents, pigs and insects. Improper disposal of broken eggs, wet bedding and other fomites contributed to Salmonella contamination. Conclusions: The results of the study indicate possibility of salmonellosis (zoonosis) in Jamaica, although the prevalence of Salmonella spp was low, and the need for improved quality of the food industry, animal care and human health to prevent salmonellosis.

Summary: Staphylococcus aureus continues to pose major public health challenges in many areas because of antibiotic resistance problems. In the Caribbean, especially Trinidad and Tobago, the challenge is not different. This study was performed to evaluate the antimicrobial resistance gene prevalence among S. aureus isolates in Trinidad and Tobago.Standard and molecular microbiological methods, including the Microscan automated system, DNA microarray and multi locus sequence typing (MLST) analysis, were performed on 309 clinical S. aureus isolates recovered from patients who were treated at three of the country's main health institutions.S. aureus exhibited susceptibilities ≥80% to eleven of the 19 antimicrobials tested against it, and these belong to the most commonly used and available antibiotics in the country. While the antibiotic to which it was most susceptible of the commonly used antibiotics was trimethoprim/sulfamethoxazole, the antibiotics to which it was least susceptible or most resistant to were ampicillin and penicillin. S. aureus isolates from the pediatric ward produced the greatest rate of susceptibility among the isolates recovered from patients admitted into hospitals, while isolates from Accident and Emergency rooms displayed the greatest susceptibilities among patients from the community.S. aureus isolates from the country did not harbor acquired resistant genes targeting clindamycin/macrolides (ermB), linezolid (cfr) or vancomycin (vanA). The blaZ gene, which is the most common beta lactam (Penicillinase) resistance mechanism for S. aureus, was observed in 88.7% of the methicillin susceptible S. aureus, while methicillin resistance mediated by the mec gene was present in 13.6%. Most of the resistance markers found in MRSA isolates were significantly associated with the ST239-MRSA-III strain in this study, and all isolates that belonged to the USA300 strain, which additionally encoded both the PVL gene and ACME cluster, belonged to CC8.Several resistant genes, such as vanA, cfr and ermB, mediating resistance in S. aureus, are currently non-existent in Trinidad and Tobago. However, the majority of SCCmec genes were observed, suggesting that there is ongoing nosocomial transmission with minimal community transmission. This calls for stringent antibiotic stewardship and policies in the country. Keywords: Staphylococcus aureus, MSSA, MRSA, Trinidad and Tobago, Clonal complexes, MLST, PVL, ST239-MRSA III, ST8-MRSA IV, USA300

Serum interleukin (IL)-17 and IL-23 and their association with systemic lupus erythematosus (SLE) is not well understood. A prospective cross sectional and observational study was done using twenty eight (28) SLE patients and fourteen (14) non SLE control subjects. Blood samples were taken to delineate the presence of IL-17 and IL-23 using ELISA among patients of the Rheumatology out-patient clinic at the San Fernando General Hospital, Trinidad and Tobago. The Systemic lupus erythematosus disease activity index (SLEDAI) 2000 score was used for evaluation of activity of the disease. It was found that of all the SLE patients 13 patients (47%) were of Afro-Trinidadian descent, 11 patients (39%) were of Indo-Trinidadian descent and 4 patients (14%) were of mixed descent. The majority of patients were within the 31-35 year age group. Serum IL-17 and IL-23 levels were greater in SLE patients compared to healthy subjects and their levels showed a positive correlation to SLEDAI score. However further experiments are needed to elucidate the role of IL-17 and IL-23 in SLE patients from Trinidad and Tobago.

Geographic spread of vancomycin-resistant enterococci (VRE) clones in cities, countries, or even continents has been identified by molecular techniques. This study aimed at characterizing virulent genes and determining genetic relatedness of 45 VRE isolates from Trinidad and Tobago using molecular tools, including polymerase chain reaction, pulsed-field gel electrophoresis (PFGE), and Random Amplification Polymorphic DNA (RAPD). The majority (84%) of the isolates wereEnterococcus faeciumpossessingvanA gene while the rest (16%) wereEnterococcus faecalispossessingvanB. Theespgene was found in all 45 VRE isolates whilehylgenes were found only inE. faeciumspecies. TheE. faeciumspecies expressed five distinct PFGE patterns. The predominant clones with similar or common patterns belonged to clones one and three, and each had 11 (29%) of the VRE isolates. Plasmid content was identified in representative isolates from each clonal group. By contrast, theE. faecalisspecies had one PFGE pattern suggesting the presence of an occult and limited clonal spread. The emergence of VRE in the country seems to be related to intra/interhospital dissemination of an epidemic clone carrying thevanA element. Therefore, infection control measures will be warranted to prevent any potential outbreak and spread of VRE in the country.

Summary Objectives There are currently 94 known pneumococcal capsular polysaccharide serotypes and their prevalence differs by geographic region and the period studied. Streptococcus pneumoniae infections have been diagnosed clinically in Trinidad and Tobago and other Caribbean countries, however data on the serotype and sequence type distributions in this country are limited. The objective of this study was to determine serotypes and multilocus sequence types (MLSTs) of invasive and non-invasive pneumococcal isolates from Trinidad and Tobago. Methods Ninety-eight pneumococcal isolates from several regional hospitals in the country were analyzed using both standard microbiological methods and molecular analysis. These isolates included invasive ( n =83) and selected non-invasive ( n =15) strains recovered before ( n =25) and after ( n =73) the introduction of the pneumococcal conjugate vaccine. Results More than half of the isolates (54.1%) were recovered from children under 15 years of age, with the largest proportion being from children under 2 years of age (24.5%). The most prevalent serotypes were 19F (18.4%), 6B (15.3%), 23F (14.3%), 3 (11.2%), 19A (6.1%), 6A (5.1%), 14 (5.1%), and 9V (4.1%). The most common serotype/MLST combinations were 6B/ST138 ( n =10, 10.2%), 3/ST180 ( n =5, 5.1%), 23F/ST629 ( n =5, 5.1%), 19F/ST8398 ( n =4, 4.1%), and three each of 6B/ST145, 14/9V/ST156, 9V/ST162, 19A/320, and 3/ST10440. Conclusions This report provides the first glimpse of the prevailing pneumococcal sequence types in the country. Most of the isolates represented serotypes in the 10-valent (61.2% of isolates) and 13-valent (83.7%) pneumococcal conjugate vaccines. A detailed population study is warranted to fully determine the circulating pneumococcal sequence types. Furthermore, the implementation of an effective and continuous surveillance system in Trinidad and Tobago is paramount to monitor vaccine impact.

OBJECTIVE: To compare the QuantiFERON®-TB Gold (QFT-G) assay and tuberculin skin test (TST) in screening/diagnosis of latent tuberculosis infection (LTBI) among individuals in Trinidad & Tobago at high risk for TB. METHODS: A total of 560 individuals (TB patient contacts, HIV patients, health care workers, prison inmates, and TB patients [controls]) were recruited for the study. Blood was drawn and processed using the QFT-G assay, followed by immediate administration of TST solution on subjects' forearm. Data were analyzed with Epi InfoTM 3.5.1 software. Results were compared across the target groups using the chi-square test (P < 0.05). RESULTS: The QFT-G assay detected LTBI in 51% of the subjects (with most positive results occurring among the control group) whereas the TST detected it in 39.4% (P = 0.001). Overall, the QFT-G assay detected LTBI more frequently than the TST among all subjects except the control group, where detection favored the TST. The QFT-G assay produced indeterminate and nonreactive results in some HIV patients but required less turnaround time than the TST (23.3 h versus 70.2 h; P < 0.0001). The TST cost less per subject than the QFT-G assay (US $3.70 versus US $18.60; P = 0.0008). CONCLUSIONS: The QFT-G assay cost more but had a higher detection rate among most target groups and required less turnaround time than the TST. However, its sensitivity was lower among immunocompromised subjects. Therefore, the QFT-G assay should be used with caution for LBTI screening/diagnosis in resource-poor, high-HIV prevalence settings such as Trinidad & Tobago.OBJETIVO: Comparar la prueba QuantiFERON®-TB Gold (QFT-G) con la prueba cutánea de la tuberculina (PPD) para el tamizaje y diagnóstico de la infección tuberculosa latente (ITBL) en personas con alto riesgo de tuberculosis en Trinidad y Tabago. MÉTODOS: Para el estudio, se reclutó un total de 560 individuos (personas en contacto con pacientes de tuberculosis, pacientes con VIH, trabajadores de la salud, presidiarios y pacientes de tuberculosis [grupo testigo]). Las muestras de sangre se extrajeron y procesaron utilizando la prueba QFT-G, seguida de la aplicación inmediata de la solución de PPD en el antebrazo de las personas. Los datos se analizaron con el software Epi InfoTM 3.5.1. Los resultados obtenidos en los grupos destinatarios se compararon utilizando la prueba de la ji al cuadrado (P < 0,05). RESULTADOS: La prueba QFT-G detectó la infección tuberculosa latente en 51% de los individuos (la mayoría de los resultados positivos se presentaron en el grupo testigo) mientras que la prueba PPD la detectó en 39,4% (P = 0,001). En términos generales, la prueba QFT-G detectó la infección tuberculosa latente con mayor frecuencia que la PPD en todos los individuos, excepto en aquellos del grupo testigo, donde el índice de detección favoreció a la PPD. La prueba QFT-G produjo resultados indeterminados y no reactivos en algunos pacientes con VIH, pero requirió menos tiempo de respuesta que la PPD (23,3 h contra 70,2 h; P < 0.0001). La prueba PPD tuvo un costo menor por individuo que la QFT-G (US$3,70 en comparación con US$18,60; P = 0.0008). CONCLUSIONES: La prueba QFT-G tuvo un costo más elevado, pero la tasa de detección fue más alta en la mayoría de los grupos destinatarios y el tiempo de respuesta fue más rápido en comparación con la PPD. Sin embargo, la sensibilidad de la prueba QFT-G fue inferior entre los individuos inmunodeficientes. Por consiguiente, se deben tomar las precauciones necesarias para utilizar la prueba QFT-G en el tamizaje y diagnóstico de ITBL en entornos de escasos recursos y alta prevalencia de VIH como Trinidad y Tabago.

We report on a fine molecular and phylogenetical characterization of circulating Mycobacterium tuberculosis strains isolated from patients during a 1-year period in Trinidad and Tobago (TT). The spoligotyping data coupled to minisatellite typing and available epidemiological data showed that a single major clone of "evolutionary modern" tubercle bacilli (SIT566) was responsible for more than half of the tuberculosis (TB) cases. It preferentially infected younger age groups (mean 39.1 years versus 47.7 years for other genotypes, p0.0005), and was overrepresented in Port-of-Spain (1 out of 3 patients). A comparison of genotyping results to data gathered for 6 Caribbean countries (n=2653 clinical isolates) showed that the overall lineage distribution in TT was completely different from its neighbors, e.g., TT was the only country harboring a unique sublineage of the Latin AmericanMediterranean (LAM) family, designated LAM-10CAM with phylogeographical specificity for Cameroon and neighboring countries in West Africa; interestingly 3/4 of the patients within this group in TT were African descendants. Similarly, strains belonging to East African Indian (EAI) lineage with phylogeographical specificity for the Indian subcontinent, were found in TT (13% of all strains), but were absent among the neighboring countries. Although the predominant SIT566 was not yet detected elsewhere in the Caribbean, available information underlined that this genotype was already present in the United States as imported cases of disease among TT-born patients. Characterization of SIT566 strains using 12-, 15- and 24-loci MIRU typing, and comparison of results to international databases showed that these isolates were characterized by a common 12-loci MIRU pattern 224315153324 corresponding to MIRU International Type-MIT633 in 21/25 strains tested, as well as its 4 variants; an orphan pattern , MIT27-, MIT117-, and MIT1158-. Extended 24-loci MIRU typing led to a predominant pattern 224315153324323483334323 in a total of 16/21 MIT633 isolates, as well as identification of 3 supplemental patterns. Comparison of 24-loci MIRU data with the international database MIRU-VNTRplus showed the unique nature of the patterns obtained in TT. Further analysis using the Levenshtein algorithm showed that the first 2 closest matches with the SIT566/MIT633 clone belonged to the X lineage strains in MIRU-VNTRplus. This observation corroborates our preliminary spoligotyping-based analysis using minimum spanning and neighbor-joining trees, which suggested a phylogenetical relatedness of the SIT566 clone with SIT119, which represents X1 lineage prototype in SpolDB4 database. We hypothesize that the predominant SIT566 clone might have evolved from a pool of X lineage M. tuberculosis strains with phylogeographical affinity for Anglo-Saxon descendants.

This report is based on a 1-year recruitment of all of the culture-positive Mycobacterium tuberculosis cases in Trinidad and Tobago ( n = 132). The study population was characterized by a high male-to-female sex ratio of 4 and a human immunodeficiency virus-tuberculosis (TB) coinfection rate of 30%. It mainly occurred among African descendants, who represent 37.5% of the total population but 69.7% of all TB cases ( P < 0.001). Spoligotyping resulted in 25 different patterns and 12 clusters (2 to 74 strains per cluster), with the predominance of a highly conserved spoligotype international type clone, SIT566.

Background: There are variations in the epidemiology, prevalent pathogens and antimicrobial susceptibility patterns of infections in the intensive care unit (ICU) from one health care facility to another, hospital to hospital, and country to country. This study was undertaken to determine and document the frequency of occurrence of microbial isolates and their antibiotic susceptibility pattern from clinical specimens received from the ICU of a tertiary regional hospital in Trinidad and Tobago. Materials & methods: Microbial isolates from patients admitted to the ICU of the Eric Williams Medical Sciences Complex over a 4-year period were investigated. Automated systems and Standard microbiological methods including BACTEC 9240 (Becton-Dickinson Microbiology Systems), MicroScan Walk Away 96 SI (Dade Behring, USA), modified Kirby Bauer disc diffusion and Etest were used. Clinical specimens from 1,128 patients admitted to the ICU during the study period were processed, and 869 pathogens were recovered from 638 positive cultures. Results: The most frequent pathogens were recovered from respiratory tract specimens, while the Enterobacteriaceae groups of organisms were the most prevalent isolates. Except for Acinetobacter species that exhibit a consistent multiple drug resistant patterns, all the pathogens showed variable susceptibility to the readily available antimicrobials in the country. A 4.2% incidence rate of ESBL producers was encountered among the K. pneumoniae and E. coli isolates from the unit. Methicillin-resistant S. aureus was noted to be on the decline in this unit, but we observed the emergence of genuine vancomycin resistant methicillin-resistant S. aureus.Conclusions: Although Enterobacteriaceae and Pseudomonas aeruginosa were the most frequent isolates, there are still sufficient treatment options for patients infected with these organisms in the unit. Continuous surveillance and monitoring for multiple drug resistant pathogens in the unit should still be paramount especially with the ongoing establishment of the National Oncology Center and National Organ Transplant Units at the complex. There is an equal need for further studies on the determinants of drug resistance in this unit.

Summary Objectives Methicillin-resistant Staphylococcus aureus (MRSA), first reported in a British hospital in the early 1960s, has now reached global proportions. Geographic spread of one or several MRSA clones in a city, country, and even among countries and continents has been identified by molecular techniques. We sought to determine whether clonal spread of MRSA has occurred in Trinidad and Tobago from all MRSA isolates collected between 2000 and 2001. Methods Clinical isolates of MRSA from three major hospitals in Trinidad and Tobago were identified by standard laboratory methods and analyzed using multiplex polymerase chain reaction (PCR) and pulsed-field gel electrophoresis (PFGE) after Sma I digestion. Results There was a 12.8% prevalence of MRSA in three major regional hospitals in Trinidad and Tobago. All 60 randomly selected MRSA strains from these hospitals produced similar PFGE banding patterns, suggesting a genetic relatedness among strains and that they belonged to a single clonal family. All isolates were negative for the Panton–Valentine leukocidin gene ( pvl ). These strains shared a PFGE banding pattern approximately (96%) the same as a Canadian strain called CMRSA-6 in the Canadian National Microbiology Laboratory database. Conclusions We conclude that only one major PFGE genotype of MRSA clone is circulating among the three major regional hospitals in Trinidad and Tobago suggesting one of three possible scenarios of microevolution: (1) all were from the dissemination of a single epidemic MRSA clone prevailing in these hospitals in Trinidad and Tobago; or (2) MRSA in Trinidad and Tobago is evolving more slowly than in other countries; or (3) that if other MRSA clones have been present in Trinidad and Tobago, they have not persisted.

Background: The objective of the study was assessment of knowledge, attitudes, and practices (KAP) towards infection prevention among healthcare workers (HCW) in Trinidad and Tobago.Methods: A cross-sectional study was conducted among 300 HCWs from three regional hospitals in Trinidad and Tobago about their knowledge, attitude, and practice towards infection prevention in the country. All information’s were collected through interview using structured questionnaires. The data was collected from April to June 2016; thoroughly checked and cleaned for completeness before analysis with SPSS version 20 statistical software. The 95% confident interval and the p value were used to check for association between the dependent and independent variables. A p value of

With the exception of some French-speaking islands, data on tuberculosis (TB) in the Caribbean are scarce. In this study, we report a first assessment of genetic diversity of a convenience sample ofMycobacterium tuberculosisstrains received from twelve Caribbean territories by spoligotyping and describe their drug-resistance patterns. Of the 480 isolates, 40 (8.3%) isolates showed resistance to at least one anti-TB drug. The proportion of drug-resistant strains was significantly higher in The Bahamas (21.4%;P=0.02), and Guyana (27.5%;P<0.0001), while it was significantly lower in Jamaica (2.4%;P=0.03) than in other countries of the present study. Regarding genetic diversity, 104 distinct spoligotype patterns were observed: 49 corresponded to clustered strains (2 to 93 strains per cluster), while 55 remained unclustered among which 16 patterns were not reported previously. Combining the study results with regional data retrieved from the international SITVIT2 database underlined a connection between frequency of certainM. tuberculosisphylogenetic lineages and the language spoken, suggesting historical (colonial) and ongoing links (trade, tourism, and migratory flows) with European countries with which they shared a common past.