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Toxoplasma gondii is a zoonotic parasite of worldwide distribution. The consumption of infected pork meat has been suggested to be an important source for human infection in the tropical area of Yucatan, Mexico. We performed a cross-sectional study of 12 farms across the state to investigate the seroprevalence of Toxoplasma gondii infection in domestic pigs. In total, 632 samples were obtained from 2 different environmental zones (tropical deciduous low forest and tropical sub-deciduous medium forest) and 2 abattoirs. The modified agglutination test (MAT) was used to assess the seroprevalence of T. gondii in pigs and to evaluate 2 globally used serological tests, the Dye test (DT) and ID Screen® ELISA multi-species, and a commercial ELISA kit (Human Toxo IgG, Human-diagnostics), which is widely used locally in this geographical area. The overall prevalence obtained with the MAT (cut-off ≥1:25) among the 632 pigs was 1.4% (95% CI, 0.6–2.7%). The seroprevalence obtained for the different age groups was 0.6%, 0.7%, 1.8%, and 6.8% among 2–3, 3–4, 4–5, and ≥5-mo-old pigs. This increase in the seroprevalence was statistically significant for the 2 older groups (odds ratio [OR] 3.9–7.1, P < 0.05) in comparison with younger groups. DT at >4 IU dilution had a perfect agreement and 100% of sensitivity and specificity when compared with the MAT. Although ID Screen® had only a fair agreement (κ = 0.389) with the MAT, the McNemar test showed that the results of these tests were comparable (P = 0.29). The Human Toxo ELISA showed no agreement with MAT, ID Screen®, and DT (κ = 0.000–0.023, McNemar P < 0.05). This ELISA was lacking in specificity, accuracy, and precision; hence, we do not recommend its use for T. gondii diagnosis in pig serum.

The ongoing outbreaks of mpox highlight the urgent need for a rapid and low-cost diagnostic test to accurately detect and control this emerging disease. We estimated the analytical sensitivity using viral culture of the monkeypox virus clade IIb lineage B1 and clinical diagnostic performance of 3 antigen detection rapid diagnostic tests (Ag-RDT) by using skin swab samples and upper-respiratory swab samples from mpox patients in the Democratic Republic of the Congo and the United Kingdom. The analytical limit of detection was 1.0 × 10 plaque-forming units/mL, fulfilling World Health Organization recommendations. Specificity of the 3 Ag-RDTs was 100%, but sensitivity was estimated at 0.00%-15.79% using skin samples and 0.00% using respiratory samples. None of the 3 Ag-RDTs reached the World Health Organization's target clinical sensitivity, and we do not recommend them as diagnostic or screening tools for suspected mpox cases. Accurate Ag-RDTs for mpox diagnosis remain urgently needed.

IntroductionHigh costs of screening and diagnostic tests remain a major barrier to timely tuberculosis (TB) identification in resource-limited settings. Evidence on the cost-effectiveness of scalable screening algorithms is limited. Start4All is a research project aimed at developing and evaluating algorithmic approaches to TB screening and diagnosis, with the goal of optimising technical and allocative efficiency when expanding diagnostic coverage to primary healthcare and community settings.Methods and analysisFive screening and diagnostic tests will be evaluated: a capillary blood-based assay (C-reactive protein (CRP)), sputum-based rapid molecular tests (PCR; individual and pooled Xpert MTB/RIF Ultra assay (Xpert Ultra, Cepheid®, California, USA)), a lateral-flow urine-based test for lipoarabinomannan (LF-LAM), and digital chest X-rays with artificial intelligence-based computer-aided detection (CXR-CAD). A microbiological reference standard of positive culture using the mycobacteria growth indicator tube will be used to confirm TB disease.We will compare the cost and effectiveness of concurrent and sequential positive serial combinations (screening algorithms) of CRP, CXR-CAD, LF-LAM, individual and pooled Xpert Ultra. Diagnostic performance will be estimated using sensitivity, specificity, predictive values and proportions of positive results, with Bayesian inference used to derive these estimates. The analysis will include adults (15 years and older) only and will be stratified by HIV status and level of care, including facility and community-based case finding. Effectiveness will be assessed based on the number of people with TB detected. Cost analysis will be conducted from the provider perspective, incorporating commodity and implementation costs. A decision tree model will be developed to assess the cost per number of persons with confirmed TB detected across all countries. Probabilistic sensitivity analysis will be conducted to account for uncertainty in model parameters, incorporating willingness-to-pay and willingness-to-accept thresholds.Ethics and disseminationWHO ethical review committee approval ERC.0003921. Data will be available on reasonable request to the principal investigator of the consortium.Trial registration numberNCT05845112.

Toxoplasmosis is a worldwide distributed parasitic disease caused by the zoonotic pathogen Toxoplasma gondii. Serological studies have estimated that more than 30% of the human population has been exposed to this protozoan. T. gondii is considered a leading cause of death attributed to foodborne illness. The consumption of infected pork meat is suggested to be an important source for human infection. However, the prevalence of T. gondii in pigs vary greatly between countries, the reasons for this heterogeneity has been addressed to the differences in climate distribution, environments, husbandry systems and fam management. The geographical location of this study is Yucatan, a state located in the south-east of Mexico. Yucatan is considered an endemic area of toxoplasmosis; the last National Seroepidemiological Health Survey (NSHS) revealed more than 70% of prevalence among the human population. Numerous studies suggested that the consumption of pork in this geographical area may be a major source of T. gondii. The aims of this study were to investigate the disease levels in the pig industry of Yucatan, assess an in-home ELISA widely used in this area (ELISA kit Human- GmbH, WB), study the risk factors associated with the disease in theses pig farms and evaluate the contamination with T. gondii in pork meat intended for human consumption. To do that, swine blood samples were collected through a cross-sectional age stratified opportunistic sampling of 12 farms across the state during the year 2014. Farm management and characteristics were obtained by interviewing farmers. In addition, meat and blood samples were collected from a local abattoir from 2013 to 2015. Anti - T. gondii IgG antibody levels were investigated with the well validated MAT (Modified Agglutination Test), with an ELISA (Enzyme-Linked Immunosorbent Assay) for use on pig serum (ID Screen ®, IDVet) and with the gold standard Dye test. The overall seroprevalence was 1.4% (95%CI: 0.6%-2.7%) among 632 pigs sampled. This seroprevalence increased with age (p < 0.05), reaching the 11.5% (95% CI: 2.5%-30%) in pigs older than 20 weeks. The seroprevalence was even higher, 17.8% (95% CI: 6%-37%), in slaughtered animals (n=34). In addition, T. gondii prevalence was investigated using a highly sensitive nested PCR protocol targeting the SAG1 gene. PCR diagnosis revealed the high frequency of 21.2% (CI: 18%-24.6%) of T. gondii DNA circulating in the blood of these pigs (n=632). Furthermore, MLST (Multi-Locus Sequence Typing) of four alleles (SAG1, SAG2, SAG3 and GRA6) allowed, for first time in Yucatan, genetic diversity to be assessed.Data revealed the presence of high genetic diversity among T. gondii strains of this geographical area with shared alleles to strains from both North and South/Central America origin. Moreover, a relatively high number of pigs presented multiple infections with different T. gondii strains suggesting high levels of T. gondii transmission on the intensive pig farms sampled. The frequency of T. gondii DNA was also investigated in pig tongues sampled at the abattoir of which 38.2% (95% CI: 22%-56.4%) were shown to harbour T. gondii DNA in their tissue. The viability of the parasite was also investigated in the tongues intended for human consumption and a total of 8.8% (95% CI: 1.8%-23%) were shown to have viable T. gondii using a mouse bioassay.

Background Clinical cases of Erysipelothrix rhusiopathiae, a zoonotic gram-positive bacterium, have been reported in many ruminant species, including in cattle, deer, moose and muskoxen. Fatal cases have been repeatedly reported in cattle over the years but to date there is only one Japanese study investigating the seroprevalence of this bacterium in cattle using the growth agglutination test (GAT). This technique is subjective, time-consuming, expensive and hazardous compared to modern serological tests such as enzyme-linked immunosorbent assays (ELISA) or the newly developed fluorescent microbead-based immunoassays (FMIA). Results A FMIA based on the surface protein SpaA (rSpaA415) antigen of E. rhusiopathiae developed in this study had an almost perfect agreement with the GAT (k=0.83) and showed a sensitivity of 89.7% and a specificity of 92.9% when compared to the GAT. Overall, detection rates of E. rhusiopathiae antibody positive samples were 13.8% (51/370) in British herds and 6% (12/200) in US herds. Positive cattle were present in 34.3% (24/70) of the investigated British farms and in 34.7% (8/23) of the US farms with an on-farm prevalence of 7.1% to 100% for the British and 8.3-30% for the US. Conclusions FMIA is a fast, safe and economic alternative to the GAT for the diagnosis of E. rhusiopathiae in cattle. This work is the first seroprevalence study of E. rhusiopathiae in healthy farmed cattle in the Great Britain and the US and revealed that infection occurs at a low level. Further investigations to evaluate risks of zoonotic transmission when handling cattle are needed.