Evaluating the ability of non-tuberculous mycobacteria to induce non-specific reactions in bovine tuberculosis diagnostic tests in guinea pigs and cattle.

Limitations in diagnostic test performance are one of the major challenges hampering the eradication of bovine tuberculosis (bTB). Non-tuberculous mycobacteria (NTM) are considered one of the main causes of non-specific reactions in the intradermal tuberculin test, the most widely used bTB diagnostic test. To determine the role of NTMs in bTB misdiagnosis in Spain, an experimental study including the NTM species most commonly found in bTB-positive animals from bTB-free farms in the country (M. avium subsp. avium (Maa), "Mycobacterium avium subsp. hominissuis" (Mah), M. bourgelatii, M. intermedium, M. kansasii and M. nonchromogenicum) was carried out on guinea pigs and cattle. First, guinea pigs were sensitized with the selected NTMs, and six weeks post-sensitization four antigen mixtures (bovine-PPD, avian-PPD, P22 and ESAT6-CFP10) were inoculated intradermally and their effect was measured 24- and 48-h post-inoculation. Larger erythematous reactions were observed in guinea pigs sensitized with Mah, M. kansasii, and Maa, with significant differences in the reactions measured at the bovine-PPD inoculation site for the two first bacteria compared with other NTMs. The sensitization process was repeated in cattle, and five months post-sensitization the same antigen mixtures were inoculated in the cervical region and responses were measured at 48- and 72-h post-inoculation. A significantly higher increase in the skinfold thickness measured at the bovine-PPD inoculation site was observed in calves sensitized with Mah, Maa, M. intermedium and M. kansasii. These results demonstrate that certain NTM species may play a more significant role in bTB diagnostic interferences and show that results obtained in guinea pig and bovine models do not always coincide.

Non-tuberculous mycobacteria: occurrence in skin test cattle reactors from official tuberculosis-free herds.

Non-tuberculous mycobacteria (NTM) are considered a relevant cause of non-specific reactions to the most widely applied bovine tuberculosis (bTB) test, the intradermal tuberculin test. In order to establish which NTM species might act as a potential source of such diagnostic interference, a collection of 373 isolates obtained from skin test positive cows from 359 officially tuberculosis-free (OTF) herds, culled in the framework of the bTB eradication campaign in Spain, were identified at the species level through PCR and Sanger sequencing of the 16S rDNA, hsp65 and rpoB genes. Of the 308 isolates for which a reliable identification was achieved, 32 different mycobacterial species were identified, with certain species being most represented: among M. avium complex members (n = 142, 46.1%), M. avium subsp. hominissuis (98; 69.0%) was the most abundant followed by M. avium subsp. avium (33, 23.2%), and M. intracellulare (7, 4.9%). Among non-MAC members (n = 166, 53.9%), M. nonchromogenicum (85; 27.6%) and M. bourgelatii (11; 5.6%) were the predominant species. In addition, mixed results were obtained in 53 isolates presenting up to 30 different genotypes, which could be indicative of new mycobacterial species. Our results represent a first step toward characterizing the diversity of NTM species that could interfere with official diagnostic tests for bTB eradication in Spain.

The Effectiveness of an Intervention Programme for Reducing Peer Rejection in Early Childhood Education.

In the early years of schooling, peer groups are key to fostering students' overall learning and development. Yet it has been found that around 10% of children suffer from peer rejection in the classroom, with this problem entailing negative consequences both in the short and long term. The problem proves difficult for adults to detect, which usually leads to a delay in measures being taken to intervene and prevent it. This study applies an experimental design with pre-test and post-test measurements in two groups-experimental and control-in order to address the problem of rejection in the early years of schooling. It explores aspects such as sociometric type, degree of victimisation, social and antisocial behaviour, as well as problematic situations among 637 students at six schools. We then implement an intervention programme for socioemotional competence throughout a school year in an effort to improve students' social skills and relationships, focusing specifically on preventing and reducing the rejection experienced by some of these children. The programme comprises 35 teaching activities and strategies that promote the development of competences for student inclusion (curbing aggression, encouraging teamwork, fostering self-esteem, etc.) and that seek to involve all students, teachers, and relatives by offering an array of complementary resources that enrich the initiatives applied (a programme calendar, assessment notebook, questionnaires, related website, material resources). After the intervention programme, it was found that the experimental group had reduced the percentage of students who suffered rejection from 9.9% to 7.3%, although the same was not true of the control group, which went from 9.5% of rejected students to 10.2%. The reduction in the percentage of rejected students in the experimental group after the application of the programme is an encouraging result that invites us to continue working on more comprehensive interventions to prevent and reduce this phenomenon.

Risk Factors for Salmonella Detection in Commercial Layer Flocks in Spain.

Trends in Salmonella human infections are assumed to be related to the distribution of the pathogen in the animal reservoir/food products, and cases in humans are most often linked to poultry and poultry products (eggs, meat). Therefore, ongoing Salmonella national control programs (NCPs) in European Union Member States have the objective of monitoring and reducing its prevalence in commercial poultry flocks. Results from NCPs have shown certain factors (housing systems, season of sampling and if sampling is conducted by food business operators (FBOps) or competent authorities (CAs), among others) can influence detection rates, but associations are often not consistent. Here, we analyzed data from the Spanish NCP on 7216 laying hen flocks subjected to 36,193 sampling events over a six-year period to characterize its performance and identify variables influencing detection rates. Overall, 1205 sampling events were positive for Salmonella spp. (any serovar) and 132 for S. Enteritidis-S. Typhimurium/monophasic. Bayesian multivariable models adjusting for multiple covariates concluded that sampling events later in the year, in caged flocks with older animals and conducted by CAs had increased odds of positivity for Salmonella spp., revealing aspects linked with a differential estimation of Salmonella levels in laying hen flocks.

Evaluation of the performance of the IFN-γ release assay in bovine tuberculosis free herds from five European countries.

The diagnostic methods for granting and maintenance of the official tuberculosis-free (OTF) status and for intra-Community movement of cattle are the tuberculin skin tests (single or comparative) and the interferon-γ (IFN-γ) release assay (IGRA). However, until now, IGRAs have been primarily applied in infected farms in parallel to the skin test to maximize the number of infected animals detected. Therefore, an evaluation of the performance of IGRAs in OTF herds to assess whether if their specificity is equal to or higher than that of the skin tests is needed. For this, a panel of 4365 plasma samples coming from 84 OTF herds in six European regions (five countries) was assembled and analysed using two IGRA kits, the ID Screen® Ruminant IFN-g (IDvet) and the Bovigam™ TB Kit (Bovigam). Results were evaluated using different cut-offs, and the impact of herd and animal-level factors on the probability of positivity was assessed using hierarchical Bayesian multivariable logistic regression models. The percentage of reactors ranged from 1.7 to 21.0% (IDvet: S/P ≥ 35%), and 2.1-26.3% (Bovigam: ODbovis-ODPBS ≥ 0.1 and ODbovis-ODavium ≥ 0.1) depending on the region, with Bovigam disclosing more reactors in all regions. The results suggest that specificity of IGRAs can be influenced by the production type, age and region of origin of the animals. Changes in the cut-offs could lead to specificity values above 98-99% in certain OTF populations, but no single cut-off yielding a sufficiently high specificity (equal or higher than that of skin tests) in all populations was identified. Therefore, an exploratory analysis of the baseline IFN-γ reactivity in OTF populations could help to assess the usefulness of this technique when applied for the purpose of maintaining OTF status.

Once bitten twice shy: Risk factors associated with bovine tuberculosis recurrence in Castilla y Leon, Spain.

Persistence of bovine tuberculosis (bTB) in cattle herd remains a major challenge in disease elimination due to the ineffectual removal of all infected animals in a bTB breakdown. Characterization of herds with a higher probability of experiencing further bTB breakdowns can help to implement specific risk-based policies for disease control and eradication. Here, our aim was to identify herd- and breakdown-level risk factors in bTB infected herds in Castilla y Leon, Spain, associated with a decreased time to recurrence and an increased risk of recurrence using a mixed effects Cox proportional hazards model and a multivariable logistic regression model, respectively. Results revealed that location (province), herd size and number of incoming animals/contacts were good predictors of a decreased time to bTB recurrence and an increased risk of becoming a recurrent herd. Additionally, the duration of the previous outbreak and the number of IFN-γ herd-tests applied in it were associated with increased odds of (an early) recurrence. Risk factors identified here can be used for early identification of herds in which bTB eradication may be more challenging and that should thus be subjected to increased control efforts. The characterization of high-risk herds may help to minimize the risk of reinfection and emphasize early detection and removal of bTB positive animals in the herd.

First Record of Colletotrichum anthrisci Causing Anthracnose on Avocado Fruits in Chile.

Anthracnose caused by Colletotrichum species is one of the most frequent and damaging fungal diseases affecting avocado fruits (Persea americana Mill.) worldwide. In Chile, the disease incidence has increased over the last decades due to the establishment of commercial groves in more humid areas. Since 2018, unusual symptoms of anthracnose have been observed on Hass avocado fruits, with lesions developing a white to gray sporulation. Morphological features and multi-locus phylogenetic analyses using six DNA barcodes (act, chs-1, gapdh, his3, ITS, and tub2) allowed the identification of the causal agent as Colletotrichum anthrisci, a member of the dematium species complex. Pathogenicity was confirmed by inoculating healthy Hass avocado fruits with representative isolates, reproducing the same symptoms initially observed, and successfully reisolating the same isolates from the margin of the necrotic pulp. Previously, several Colletotrichum species belonging to other species complexes have been associated with avocado anthracnose in other countries. To our knowledge, this is the first record of C. anthrisci and of a species of the dematium species complex causing anthracnose on avocado fruits in Chile and worldwide.

Patterns of antimicrobial resistance in Salmonella isolates from fattening pigs in Spain.

BACKGROUND: Swine are considered a major source of foodborne salmonellosis, a public health issue further complicated by the circulation of multidrug-resistant Salmonella strains that threaten the safety of the food chain. The current study aimed to identify patterns that can help to understand the epidemiology of antimicrobial resistance (AMR) in Salmonella in pigs in Spain through the application of several multivariate statistical methods to data from the AMR national surveillance programs from 2001 to 2017. RESULTS: A total of 1,318 pig Salmonella isolates belonging to 63 different serotypes were isolated and their AMR profiles were determined. Tetracycline resistance across provinces in Spain was the highest among all antimicrobials and ranged from 66.7% to 95.8%, followed by sulfamethoxazole resistance (range: 42.5% - 77.8%), streptomycin resistance (range: 45.7% - 76.7%), ampicillin resistance (range: 24.3% - 66.7%, with a lower percentage of resistance in the South-East of Spain), and chloramphenicol resistance (range: 8.5% - 41.1%). A significant increase in the percentage of resistant isolates to chloramphenicol, sulfamethoxazole, ampicillin and trimethoprim from 2013 to 2017 was observed. Bayesian network analysis showed the existence of dependencies between resistance to antimicrobials of the same but also different families, with chloramphenicol and sulfamethoxazole in the centre of the networks. In the networks, the conditional probability for an isolate susceptible to ciprofloxacin that was also susceptible to nalidixic acid was 0.999 but for an isolate resistant to ciprofloxacin that was also resistant to nalidixic acid was only 0.779. An isolate susceptible to florfenicol would be expected to be susceptible to chloramphenicol, whereas an isolate resistant to chloramphenicol had a conditional probability of being resistant to florfenicol at only 0.221. Hierarchical clustering further demonstrated the linkage between certain resistances (and serotypes). For example, a higher likelihood of multidrug-resistance in isolates belonging to 1,4,[5],12:i:- serotype was found, and in the cluster where all isolates were resistant to tetracycline, chloramphenicol and florfenicol, 86.9% (n = 53) of the isolates were Typhimurium. CONCLUSION: Our study demonstrated the power of multivariate statistical methods in discovering trends and patterns of AMR and found the existence of serotype-specific AMR patterns for serotypes of public health concern in Salmonella isolates in pigs in Spain.

Use of Whole-Genome Sequencing to Unravel the Genetic Diversity of a Prevalent Mycobacterium bovis Spoligotype in a Multi-Host Scenario in Spain.

Despite the efforts invested in the eradication of bovine tuberculosis in Spain, herd prevalence has remained constant in the country during the last 15 years (~1.5-1.9%) due to a combination of epidemiological factors impairing disease control, including between-species transmission. Here, our aim was to investigate the molecular diversity of Mycobacterium bovis isolates belonging to the highly prevalent SB0339 spoligotype in the cattle-wildlife interface in different regions of Spain using whole-genome sequencing (WGS). Genomic data of 136 M. bovis isolates recovered from different animal species (cattle, wild boar, fallow deer, and red deer) and locations between 2005 and 2018 were analyzed to investigate between- and within-species transmission, as well as within-herds. All sequenced isolates differed by 49-88 single nucleotide polymorphisms from their most recent common ancestor. Genetic heterogeneity was geographic rather than host species-specific, as isolates recovered from both cattle and wildlife from a given region were more closely related compared to isolates from the same species but geographically distant. In fact, a strong association between the geographic and the genetic distances separating pairs of M. bovis isolates was found, with a significantly stronger effect when cattle isolates were compared with wildlife or cattle-wildlife isolates in Spain. The same results were obtained in Madrid, the region with the largest number of sequenced isolates, but no differences depending on the host were observed. Within-herd genetic diversity was limited despite the considerable time elapsed between isolations. The detection of closely related strains in different hosts demonstrates the complex between-host transmission dynamics present in endemic areas in Spain. In conclusion, WGS results a valuable tool to track bTB infection at a high resolution and may contribute to achieve its eradication in Spain.

Genomic characterization of multidrug-resistant Salmonella serovar Kentucky ST198 isolated in poultry flocks in Spain (2011-2017).

Salmonella Kentucky is commonly found in poultry and rarely associated with human disease. However, a multidrug-resistant (MDR) S. Kentucky clone [sequence type (ST)198] has been increasingly reported globally in humans and animals. Our aim here was to assess if the recently reported increase of S. Kentucky in poultry in Spain was associated with the ST198 clone and to characterize this MDR clone and its distribution in Spain. Sixty-six isolates retrieved from turkey, laying hen and broiler in 2011-2017 were subjected to whole-genome sequencing to assess their sequence type, genetic relatedness, and presence of antimicrobial resistance genes (ARGs), plasmid replicons and virulence factors. Thirteen strains were further analysed using long-read sequencing technologies to characterize the genetic background associated with ARGs. All isolates belonged to the ST198 clone and were grouped in three clades associated with the presence of a specific point mutation in the gyrA gene, their geographical origin and isolation year. All strains carried between one and 16 ARGs whose presence correlated with the resistance phenotype to between two and eight antimicrobials. The ARGs were located in the Salmonella genomic island (SGI-1) and in some cases (blaSHV-12, catA1, cmlA1, dfrA and multiple aminoglycoside-resistance genes) in IncHI2/IncI1 plasmids, some of which were consistently detected in different years/farms in certain regions, suggesting they could persist over time. Our results indicate that the MDR S. Kentucky ST198 is present in all investigated poultry hosts in Spain, and that certain strains also carry additional plasmid-mediated ARGs, thus increasing its potential public health significance.

Monitoring of Antimicrobial Resistance to Aminoglycosides and Macrolides in Campylobacter coli and Campylobacter jejuni From Healthy Livestock in Spain (2002-2018).

Antimicrobial resistance (AMR) in Campylobacter spp. (Campylobacter coli and Campylobacter jejuni) is a concern due to its importance in public health, particularly when it involves aminoglycosides and macrolides, drugs of choice for treatment of human cases. Co-resistance to these two antimicrobial classes involves transfer of genetic elements and/or acquisition of mutations in different genetic loci, which can in turn spread through vertical or horizontal gene transfer (HGT) phenomena, with each route having different potential implications. This study aimed at evaluating the association between the presence of phenotypic resistance to these two antimicrobial classes in C. coli and C. jejuni recovered from livestock at slaughterhouses in Spain (as part of the AMR surveillance program), and at assessing the genetic heterogeneity between resistant and susceptible isolates by analysing the "short variable region" (SVR) of the flaA gene. Over the 2002-2018 period, antimicrobial susceptibility test results from 10,965 Campylobacter isolates retrieved from fecal samples of broilers, turkeys, pigs and cattle were collected to compare the proportion of resistant isolates and the Minimum Inhibitory Concentrations (MICs) against six antimicrobials including gentamicin (GEN), streptomycin (STR), and erythromycin (ERY). AMR-associated genes were determined for a group of 51 isolates subjected to whole genome sequencing, and the flaA SVR of a subset of 168 isolates from all hosts with different resistotypes was used to build a Neighbor-Joining-based phylogenetic tree and assess the existence of groups by means of "relative synonymous codon usage" (RSCU) analysis. The proportion of antimicrobial resistant isolates to both, aminoglycosides and macrolides, varied widely for C. coli (7-91%) and less for C. jejuni (all hosts 0-11%). Across hosts, these proportions were 7-56% in poultry, 12-82% in cattle, and 22-91% in pigs for C. coli and 0-8% in poultry and 1-11% in cattle for C. jejuni. Comparison of the MIC distributions revealed significant host-specific differences only for ERY in C. jejuni (p = 0.032). A significant association in the simultaneous presentation of AMR to both antimicrobial classes was observed across hosts/bacterial species. The flaA gene analysis showed clustering of isolates sharing resistotype and to a lesser degree bacterial species and host. Several resistance markers associated with resistance to aminoglycosides and macrolides were found among the sequenced isolates. The consistent association between the simultaneous presentation of AMR to aminoglycosides and macrolides in all hosts could be due to the persistence of strains and/or resistance mechanisms in Campylobacter populations in livestock over time. Further studies based on whole genome sequencing are needed to assess the epidemiological links between hosts and bacterial strains.

Socio-Emotional Variables Linked to the Consumption of Drugs amongst University Students of Social Sciences: A Pilot Study.

This descriptive and transversal study, carried out on an intentional sample of 211 subjects who were split in terms of their consumption of psychoactive substances over the last month and who were aged between 18 and 28 (M = 21.36, and SD = 1.90), aimed to explore the emotional intelligence, perceived socio-family support and academic performance of university students vis-à-vis their consumption of drugs and to examine the link between them. The goal was to define university student consumer profile through a regression model using the multidimensional Perceived Social Support Scale (EMAS) and the Trait Meta Mood Scale-24 (TMMS-24) as instruments, together with academic performance and gender. The results report alcohol, tobacco, and cannabis consumption rates that are above the levels indicated by the Spanish household survey on alcohol and drugs in Spain (EDADES 2019) for the 15-34-year-old age range in Castilla y León. A certain link was observed between the consumption of substances and academic performance, although no differences were seen in academic performance in terms of consumer type. There was also no clear link observed between emotional intelligence and academic performance or between social support and academic performance. The predictive contribution of the variables included in the regression model was low (9%), which would advocate completing the model with other predictive variables until more appropriate predictability conditions can be found.

Evaluation of the performance of slaughterhouse surveillance for bovine tuberculosis detection in Castilla y Leon, Spain.

Post-mortem inspection (PMI) of routinely slaughtered cattle in abattoirs is an extremely valuable tool for detecting bovine tuberculosis (bTB) infected herds that can supplement active surveillance activities. However, its true performance is difficult to assess due to the multiple factors that may affect it. Here, we determined relative efficiencies in the detection of bTB-compatible lesions and probabilities of subsequent laboratory confirmation of abattoirs located in Castilla y Leon, one of the regions with the largest cattle population in Spain, between 2010 and 2017. The slaughtered animal population was split based on the results of the ante-mortem tests (reactors or non-reactors), and two generalized linear multivariable mixed models were fitted to each subpopulation to calculate the risk of lesion detection and laboratory confirmation per abattoir while accounting for the effect of potential confounding variables. Throughout the 8-year period, ∼30,000 reactors and >2.8 million non-reactor animals in the ante-mortem tests were culled in the abattoirs under study. Bovine TB compatible lesions were detected in 4,710 (16%) reactors and 828 (0.03%) non-reactor animals, of which >95% were confirmed as infected through bacteriology. The probability of disclosure of bTB-like lesions was associated with the animal subpopulation, type of source unit, the herd size, the year of slaughter, the breed and age of the animal, and/or the season of slaughter. The probabilities of detection of bTB-like lesions varied largely depending on the abattoir in both subpopulations, ranging from 603 to 3,070 per 10,000 animals for the reactors and 0.2-16.1 per 10,000 animals for the non-reactor animals. Results obtained here will help to quantify the performance of PMI in abattoirs in Castilla y Leon and the between-abattoir variability, and to identify animals at increased risk of having bTB-like lesions detected during PMI based on animal- and farm-related factors.

Polyresistant Mycobacterium bovis Infection in Human and Sympatric Sheep, Spain, 2017-2018.

The main etiologic agent of tuberculosis (TB) in livestock is Mycobacterium bovis; human TB cases caused by M. bovis are rare. Analysis of a TB outbreak caused by polyresistant M. bovis involving a human and sympatric sheep in Spain suggests local circulation of drug-resistant M. bovis strains among livestock.

Evaluation of Risk Factors Associated With Herds With an Increased Duration of Bovine Tuberculosis Breakdowns in Castilla y Leon, Spain (2010-2017).

The persistence of bovine tuberculosis (bTB) in certain cattle herds is a major concern in countries pursuing disease eradication worldwide. The chronic nature of the disease, the lack of performance of diagnostic tools, and the presence of wildlife reservoirs may lead infected herds to require longer periods to achieve the officially tuberculosis-free (OTF) status. Here, we evaluated the impact of farm and breakdown characteristics on the probability of disease persistence in infected farms in Castilla y Leon, a bTB-endemic region of Spain, using survival and logistic regression models. Data from bTB breakdowns occurring in 3,550 bTB-positive herds detected in 2010-2017 were analyzed. A multivariable Cox proportional hazards model was fitted using time to recover OTF status as the response variable, and a multivariable logistic regression model using the chronic status (yes/no) for herds experiencing particularly long breakdowns as the outcome variable was also used. Both analyses revealed that county-level bTB herd prevalence, herd size, number of incoming animals in the previous 3 years, number of skin test reactors in the disclosing test, and number of days between the disclosing and follow-up tests were associated with increased breakdown duration. Production type was not consistently associated with chronic infection, suggesting that once infected, it is not a significant predictor of outbreak duration beyond the initial stages of the breakdown. Province-level location and number of animals that are bacteriology-positive also affected significantly the expected herd breakdown duration, but their effect became less significant over time. Risk factors identified in this study may help to identify herds more prone to suffer chronic bTB infection that may require additional control measures early on in a breakdown.

Identifying emerging trends in antimicrobial resistance using Salmonella surveillance data in poultry in Spain.

Despite of controls and preventive measures implemented along the food chain, infection with non-typhoidal Salmonella (NTS) remains one of the major causes of foodborne disease worldwide. Poultry is considered one of the major sources of NTS. This has led to the implementation of monitoring and control programmes in many countries (including Spain) to ensure that in poultry flocks infection is kept to a minimum and to allow the identification and monitoring of circulating NTS strains and their antimicrobial resistance (AMR) phenotypes. Here, we investigated the information from the monitoring programme for AMR in Salmonella from poultry in Spain in 2011-2017 to assess the diversity in phenotypic resistance and to evaluate the programme's ability to detect multi-resistance patterns and emerging strains in the animal reservoir. Data on serotype and AMR to nine antimicrobials obtained from 3,047 NTS isolates from laying hens (n = 1,060), broiler (n = 765) and turkey (n = 1,222) recovered during controls performed by the official veterinary services and food business operators were analysed using univariate and multivariate methods in order to describe host and serotype-specific profiles. Diversity and prevalence of phenotypic resistance to all but one of the antimicrobials (colistin) were higher in NTS from broiler and turkey compared with laying hen isolates. Certain combinations of serotype and AMR pattern (resistotype) were particularly linked with certain hosts (e.g. susceptible Enteritidis with laying hens, multi-drug resistant (MDR) Derby in turkey, MDR Kentucky in turkey and broiler). The widespread presence of certain serotype-resistotype combinations in certain hosts/years suggested the possible expansion of MDR strains in the animal reservoir. This study demonstrates the usefulness of the analysis of data from monitoring programmes at the isolate level to detect emerging threats and suggests aspects that should be subjected to further research to identify the forces driving the expansion/dominance of certain strains in the food chain.

Assessment of the sensitivity of the bovine tuberculosis eradication program in a high prevalence region of Spain using scenario tree modeling.

In spite of the efforts invested to eradicate bovine tuberculosis (bTB) in cattle, the disease is still present in several developed countries, including Spain. Among the factors related with disease persistence in high prevalence areas, the lack of sensitivity of the screening test for detection of infected herds [single intradermal tuberculin (SIT) test] can play a major role. Here, a scenario tree model mimicking the diagnostic test scheme in place in the region of Castilla y Leon (Spain) was developed to estimate the probability of detecting bTB in an infected-non detected officially tuberculosis free (OTF) herd (herd sensitivity, HSe). In order to do so the probability of detecting at least one positive animal in the SIT test with/without post-mortem (detection of lesions and culture) confirmation in an infected herd was estimated using Monte Carlo simulation through @RISK (Palisade Co, NY, USA). Uncertainty on the accuracy of the diagnostic tests was introduced in the model using distributions based on the literature. The performance of the model was evaluated by comparing the predicted number of SIT/post-mortem positive animals in infected herds with those observed in newly detected bTB-infected herds in the region in 2011-2015. The estimated HSe of the SIT test was 76.2% (95% probability interval: 19.8-97.6). According to the model, bTB infection would be then confirmed through culture in 65.3% (95% PI: 50.0-82.3) of the herds detected through the SIT test, so that overall the proportion of infected-non detected OTF herds in which the infection could be confirmed after the initial SIT test was 49.6% (95% PI: 9.75-80.3). The predicted HSe of both SIT test and culture was directly correlated with herd size. Results from the model suggest a moderate but highly variable HSe of the current surveillance system in place for bTB detection in OTF herds located in high prevalence areas, that could be maximized by performing multiple tests within a year as indicated in the Spanish eradication program (with a median SIT HSe of 87% when two consecutive tests were considered). In addition, these results highlight the usefulness of performing subsequent SIT tests to rule out infection in SIT-positive herds even when the causative agent cannot be isolated.

Evaluation of the effectiveness of the surveillance system for tuberculosis in cattle in Spain.

In Spain, the eradication of tuberculosis (TB) in cattle has had some setbacks and therefore we need to re-evaluate the current surveillance strategies. The Spanish cattle TB surveillance system consists of three components: a) routine skin testing, b) slaughterhouse surveillance, and c) pre-movement controls. Our objectives were to assess the effectiveness of the whole surveillance system and evaluate the relative contribution of each of those components, both at national and at the province level. The effectiveness was estimated based on both the sensitivity of detection per year and the time until detection. Moreover, we evaluated the impact of different factors on that effectiveness. We used a stochastic model that simulated the spread of Mycobacterium tuberculosis-complex (MTC) infection within cattle herds that was modified to incorporate the three components of TB surveillance. Input data, at the province level, on cattle movements off-farm or the frequency of the routine screening were provided by the Spanish Ministry for Agriculture, Fisheries and Food. Under the current conditions, 96.1% of Spanish herds were detected within one year after their infection (i.e. mean sensitivity of TB surveillance), although that detection took on average 178 days. The surveillance system effectiveness was highly dependent on the routine skin testing, responsible for the detection of almost 90% of the infected herds, while slaughterhouse surveillance and pre-movement controls contributed only to the identification of a small proportion of infected herds. We observed substantial differences in the effectiveness of the surveillance components among Spanish provinces, although in general, the sensitivities were high. The most influential factor on the efficiency of TB detection in Spain was, by far, the frequency of routine controls, followed by the sensitivity of the test used. In a context of reduced funding for cattle TB eradication, the frequency of testing should be adapted based on risk-based surveillance strategies, i.e. efforts should focus on herds more likely to be infected, but also in herds more likely to infect other herds. While slaughterhouse surveillance seems a cost-effective strategy, the use of pre-movement testing at least in areas of low incidence should be further evaluated.

Teleophthalmology: a strategy for timely diagnosis of sight-threatening diabetic retinopathy in primary care, Concepción, Chile.

AIM: To estimate the prevalence of diabetic retinopathy (DR) in a diabetic population of the public health system. METHODS: This non-experimental, descriptive and cross-sectional study, with no direct control over the variables analysed, was carried out by the Telemedicine Unit of the University of Concepción (TELMED) and the Family Health Centers (CESFAM) of the Health Service Concepción, Chile, among a population of 7382 diabetic patients of the public health system, from October 2014 to June 2015. Two digital images for each patient's eyes were obtained and uploaded to the TELMED tele-ophthalmology platform. The images were remotely evaluated by a retina expert ophthalmologist for diagnosis. Treatment and a referral (if required) were recommended, and an individualised report for each patient was sent to the place of origin. RESULTS: The prevalence of DR in this study was 14.9%. Of these, 48.7% had mild non-proliferative DR (NPDR), 30.8% moderate NPDR, 15.9% severe NPDR, and 4.6% proliferative DR. Of the patients with DR, 17.8% were referred for fluorescein angiography, 4.3% for panretinal photocoagulation, 1.2% for vitrectomy, and 0.4% for cataract surgery. CONCLUSION: The use of telemedicine allowed an increased screening coverage for DR in diabetic patients. The different types of DR were detected in a timely manner, decreasing waiting times due to the lack of specialists in ophthalmology in the public health system.