What else in times of COVID-19? The role of minimally invasive autopsy for the differential diagnosis of acute respiratory failure in a case of kala-azar.

Visceral leishmaniasis (VL) is a chronic vector-borne zoonotic disease caused by trypanosomatids, considered endemic in 98 countries, mainly associated with poverty. About 50,000-90,000 cases of VL occur annually worldwide, and Brazil has the second largest number of cases in the world. The clinical picture of VL is fever, hepatosplenomegaly, and pancytopenia, progressing to death in 90% of cases due to secondary infections and multi-organ failure, if left untreated. We describe the case of a 25-year-old female who lived in the metropolitan area of Sao Paulo, who had recently taken touristic trips to several rural areas in Southeastern Brazil and was diagnosed post-mortem. During the hospitalization in a hospital reference for the treatment of COVID-19, the patient developed acute respiratory failure, with chest radiographic changes, and died due to refractory shock. The ultrasound-guided minimally invasive autopsy diagnosed VL (macrophages containing amastigote forms of Leishmania in the spleen, liver and bone marrow), as well as pneumonia and bloodstream infection by gram-negative bacilli.

[Visceral leishmaniasis in Xinjiang Uygur Autonomous Region during the COVID-19 pandemic: a case report].

OBJECTIVE: To perform an epidemiological investigation on a case of visceral leishmaniasis reported from Shule County, Kashi Prefecture, Xinjiang Uygur Autonomous Region in 2021, so as to provide insights into differential diagnosis of visceral leishmaniasis during the COVID-19 pandemic. METHODS: The epidemiological history of this case was collected, and the case was diagnosed for Leishmania infection with the immunochromatographic (rK39) strip test, bone marrow smear microscopy and PCR assay. RESULTS: The patient had typical clinical symptoms of leishmaniasis, including irregular fever, hepatosplenomeg- aly, low serum albumin and elevated globulin. Bone marrow smear microscopy identified L. donovani amastigotes, and both rK39 strip test and PCR assay were positive, while the case was tested negative for SARS-CoV-2. COVID-19 was therefore excluded and visceral leishmaniasis was diagnosed. Standard full-dose treatment with sodium stibogluconate was given, and no Leishmania was found on blood smears during the reexamination. No recurrence was found during the followup after discharge for hospital. CONCLUSIONS: During the COVID-19 pandemic, it is recommended to increase the perception of differential diagnosis of visceral leishmaniasis among first-contact doctors, and reinforce the capability of differential diagnosis and health education of visceral leishmaniasis among medical and healthcare institutions at all levels, to prevent missed diagnosis and misdiagnosis of visceral leishmaniasis.

Evaluation of Recombinase Polymerase Amplification assay for monitoring parasite load in patients with kala-azar and post kala-azar dermal leishmaniasis.

BACKGROUND: The potential reservoirs of visceral leishmaniasis (VL) in South Asia include asymptomatic and relapsed cases of VL, along with patients with post kala-azar dermal leishmaniasis (PKDL). Accordingly, accurate estimation of their parasite load is pivotal for ensuring disease elimination, presently targeted for 2023. Serological tests cannot accurately detect relapses and/or monitor treatment effectiveness, and therefore, parasite antigen/nucleic acid based detection assays remain the only viable option. An excellent option is the quantitative polymerase chain reaction (qPCR) but the high cost, technical expertise and time involved precludes its wider acceptability. Accordingly, the recombinase polymerase amplification (RPA) assay operated in a mobile suitcase laboratory has emerged not simply as a diagnostic tool for leishmaniasis but also to monitor the disease burden. METHODOLOGY/PRINCIPAL FINDINGS: Using total genomic DNA isolated from peripheral blood of confirmed VL cases (n = 40) and lesional biopsies of PKDL cases (n = 64), the kinetoplast-DNA based qPCR and RPA assay was performed and parasite load expressed as Cycle threshold (Ct) and Time threshold (Tt) respectively. Using qPCR as the gold standard, the diagnostic specificity and sensitivity of RPA in naïve cases of VL and PKDL was reiterated. To assess the prognostic potential of the RPA, samples were analyzed immediately at the end of treatment or ≥6 months following completion of treatment. In cases of VL, the RPA assay in terms of cure and detection of a relapse case showed 100% concordance with qPCR. In PKDL following completion of treatment, the overall detection concordance between RPA and qPCR was 92.7% (38/41). At the end of treatment for PKDL, 7 cases remained qPCR positive, whereas RPA was positive in only 4/7 cases, perhaps attributable to their low parasite load. CONCLUSIONS/SIGNIFICANCE: This study endorsed the potential of RPA to evolve as a field applicable, molecular tool for monitoring parasite load, possibly at a point of care level and is worthy of consideration in resource limited settings.

Spatio-temporal relative risks and priority areas for visceral leishmaniasis control in Brazil, between 2001 and 2020.

Visceral leishmaniasis (VL) is a pressing public health problem in Brazil. The proper implementation of disease control programs in priority areas is a challenge for healthcare managers. The present study aimed to analyze the spatio-temporal distribution and identify high risk areas of VL occurrence in the Brazilian territory. We analyzed data regarding new cases with confirmed diagnosis of VL in Brazilian municipalities, from 2001 to 2020, extracted from the Brazilian Information System for Notifiable Diseases. The Local Index of Spatial Autocorrelation (LISA) was used to identify contiguous areas with high incidence rates in different periods of the temporal series. Clusters of high spatio-temporal relative risks were identified using the scan statistics. The accumulated incidence rate in the analyzed period was 33.53 cases per 100,000 inhabitants. The number of municipalities that reported cases showed an upward trend from 2001 onward, although there was a decrease in 2019 and 2020. According to LISA, the number of municipalities considered a priority increased in Brazil and in most states. Priority municipalities were predominantly concentrated in the states of Tocantins, Maranhão, Piauí, and Mato Grosso do Sul, in addition to more specific areas of Pará, Ceará, Piauí, Alagoas, Pernambuco, Bahia, São Paulo, Minas Gerais, and Roraima. The spatio-temporal clusters of high-risk areas varied throughout the time series and were relatively higher in the North and Northeast regions. Recent high-risk areas were found in Roraima and municipalities in northeastern states. VL expanded territorially in Brazil in the 21st century. However, there is still a considerable spatial concentration of cases. The areas identified in the present study should be prioritized for disease control actions.

Prevalence of post kala-azar dermal leishmaniasis (PKDL) and treatment seeking behavior of PKDL patients in Nepal.

BACKGROUND: In Nepal, the burden of post kala-azar dermal leishmaniasis (PKDL) is not known since there is no active case detection of PKDL by the national programme. PKDL patients could pose a challenge to sustain visceral leishmaniasis (VL) elimination. The objective of this study was to determine the prevalence of PKDL and assess PKDL patients' knowledge on VL and PKDL, and stigma associated with PKDL. METHODOLOGY/PRINCIPAL FINDINGS: Household surveys were conducted in 98 VL endemic villages of five districts that reported the highest number of VL cases within 2018-2021. A total of 6,821 households with 40373 individuals were screened for PKDL. Cases with skin lesions were referred to hospitals and examined by dermatologists. Suspected PKDL cases were tested with rK39 and smear microscopy from skin lesions. An integrated diagnostic approach was implemented in two hospitals with a focus on management of leprosy cases where cases with non-leprosy skin lesions were tested for PKDL with rK39. Confirmed PKDL patients were interviewed to assess knowledge and stigma associated with PKDL, using explanatory model interview catalogue (EMIC) with maximum score of 36. Among 147 cases with skin lesions in the survey, 9 (6.12%) were confirmed as PKDL by dermatologists at the hospital. The prevalence of PKDL was 2.23 per 10,000 population. Among these 9 PKDL cases, 5 had a past history of VL and 4 did not. PKDL cases without a past history of VL were detected among the "new foci", Surkhet but none in Palpa. None of the cases negative for leprosy were positive for PKDL. There was very limited knowledge of PKDL and VL among PKDL cases. PKDL patients suffered to some degree from social and psychological stigma (mean ± s.d. score = 17.89 ± 12.84). CONCLUSIONS/SIGNIFICANCE: Strengthening the programme in PKDL case detection and management would probably contribute to sustenance of VL elimination. Awareness raising activities to promote knowledge and reduce social stigma should be conducted in VL endemic areas.

Evaluating and mitigating the potential indirect effect of COVID-19 on control programmes for seven neglected tropical diseases: a modelling study.

BACKGROUND: In line with movement restrictions and physical distancing essential for the control of the COVID-19 pandemic, WHO recommended postponement of all neglected tropical disease (NTD) control activities that involve community-based surveys, active case finding, and mass drug administration in April, 2020. Following revised guidance later in 2020, and after interruptions to NTD programmes of varying lengths, NTD programmes gradually restarted in the context of an ongoing pandemic. However, ongoing challenges and service gaps have been reported. This study aimed to evaluate the potential effect of the programmatic interruptions and strategies to mitigate this effect. METHODS: For seven NTDs, namely soil-transmitted helminths, schistosomiasis, lymphatic filariasis, onchocerciasis, trachoma, visceral leishmaniasis, and human African trypanosomiasis, we used mathematical transmission models to simulate the effect of programme interruptions on the dynamics of each of these diseases in different endemic settings. We also explored the potential benefit of implementing mitigation strategies, primarily in terms of minimising the delays to control targets. FINDINGS: We show that the effect of the COVID-19-induced interruption in terms of delay to achieving elimination goals might in some cases be much longer than the duration of the interruption. For schistosomiasis, onchocerciasis, trachoma, and visceral leishmaniasis, a mean delay of 2-3 years for a 1-year interruption is predicted in areas of highest prevalence. We also show that these delays can largely be mitigated by measures such as additional mass drug administration or enhanced case-finding. INTERPRETATION: The COVID-19 pandemic has brought infectious disease control to the forefront of global consciousness. It is essential that the NTDs, so long neglected in terms of research and financial support, are not overlooked, and remain a priority in health service planning and funding. FUNDING: Bill & Melinda Gates Foundation, Medical Research Council, and the UK Foreign, Commonwealth & Development Office.

Immunoprophylaxis using polypeptide chimera vaccines plus adjuvant system promote Th1 response controlling the spleen parasitism in hamster model of visceral leishmaniasis.

In recent years, several advances have been observed in vaccinology especially for neglected tropical diseases (NTDs). One of the tools employed is epitope prediction by immunoinformatic approaches that reduce the time and cost to develop a vaccine. In this scenario, immunoinformatics is being more often used to develop vaccines for NTDs, in particular visceral leishmaniasis (VL) which is proven not to have an effective vaccine yet. Based on that, in a previous study, two predicted T-cell multi-epitope chimera vaccines were experimentally validated in BALB/c mice to evaluate the immunogenicity, central and effector memory and protection against VL. Considering the results obtained in the mouse model, we assessed the immune response of these chimeras inMesocricetus auratushamster, which displays, experimentally, similar pathological status to human and dog VL disease. Our findings indicate that both chimeras lead to a dominant Th1 response profile, inducing a strong cellular response by increasing the production of IFN-γ and TNF-α cytokines associated with a decrease in IL-10. Also, the chimeras reduced the spleen parasite load and the weight a correlation between protector immunological mechanisms and consistent reduction of the parasitic load was observed. Our results demonstrate that both chimeras were immunogenic and corroborate with findings in the mouse model. Therefore, we reinforce the use of the hamster as a pre-clinical model in vaccination trials for canine and human VL and the importance of immunoinformatic to identify epitopes to design vaccines for this important neglected disease.

Severe visceral leishmaniasis and COVID-19 coinfection in an immunosuppressed patient.

Visceral leishmaniasis (VL) is an opportunistic disease in immunosuppressed individuals, who may present severe clinical conditions, such as the ones described in this patient. She lived in an endemic region for VL, and was possibly infected with L. (L.) infantum chagasi through the bite of a contaminated sand fly. This initial infection has triggered a pemphigus vulgaris condition by immunogenic proteins present in the mosquito's saliva. The immunosuppression caused by the use of high doses of corticosteroids to control the disease promoted a severe VL condition, with hepatosplenomegaly, thrombocytopenia and hemorrhages, requiring hospitalization and the onset of a subsequent SARS-CoV-2 infection. Due to the intensity of clinical manifestations related to VL, aggravated by COVID-19, she died two days after admission to the Clinical Hospital of Marilia Medical School (HC-Famema).

Digital strategies for health education on leishmaniasis / Estrategias digitales de educación en salud sobre leishmaniasis

La leishmaniasis es una enfermedad desatendida y endémica en localidades remotas de Perú, y existe evidencia de un alto índice de inexperiencia por parte del personal de salud que labora en zonas endémicas; asociada al diagnóstico, tratamiento y desconocimiento de protocolos nacionales e internacionales respecto a la enfermedad; lo que se traduce en un incremento de eventos adversos o una cura incompleta para los pacientes. Por otro lado, la pandemia por COVID-19, ha originado interrupción en los sistemas educativos, estimulando la aplicación de enfoques educativos a distancia. Se elaboró un programa académico de educación superior bajo la modalidad virtual, dirigido a profesionales sanitarios que laboran en áreas de riesgo o endémicas de leishmania, en el marco del eLearning, empleando tecnologías de información y comunicación (TIC) como herramientas para el aprendizaje; y se aplicó la metodología MEDESME en la planificación de herramientas digitales educativas. El producto incluyó la ficha académica del programa "leishmaniasis tegumentaria y visceral", diversos recursos de autoaprendizaje y estrategias de evaluación digitales. La aplicación del programa educativo permitiría capacitar y actualizar conocimientos a los profesionales de la salud y, en consecuencia, optimizar el diagnóstico, tratamiento y seguimiento de los pacientes afectados por las distintas manifestaciones de la leishmaniasis(AU)

Leishmaniasis is a neglected and endemic disease in remote locations in Peru, and there is evidence of a high rate of inexperience on the part of health personnel working in endemic areas; associated with the diagnosis, treatment and ignorance of national and international protocols regarding the disease; which translates into an increase in adverse events or an incomplete cure for patients. On the other hand, the COVID-19 pandemic has caused disruption in educational systems, stimulating the application of distance educational approaches. An academic program of higher education was developed under the virtual modality, aimed at health professionals who work in risky or endemic areas of leishmania, within the framework of eLearning, using information and communication technologies (ICT) as tools for learning; and the MEDESME methodology was applied in the planning of educational digital tools. The product included the academic record of the program "integumentary and visceral leishmaniasis", various self-study resources and digital assessment strategies. The application of the educational program would allow health professionals to be trained and updated and, consequently, to optimize the diagnosis, treatment and follow-up of patients affected by the different manifestations of leishmaniasis(AU)

Epidemiological features and spatial-temporal distribution of visceral leishmaniasis in mainland China: a population-based surveillance study from 2004 to 2019.

BACKGROUND: Although visceral leishmaniasis (VL) was largely brought under control in most regions of China during the previous century, VL cases have rebounded in western and central China in recent decades. The aim of this study was to investigate the epidemiological features and spatial-temporal distribution of VL in mainland China from 2004 to 2019. METHODS: Incidence and mortality data for VL during the period 2004-2019 were collected from the Public Health Sciences Data Center of China and annual national epidemic reports of VL, whose data source was the National Diseases Reporting Information System. Joinpoint regression analysis was performed to explore the trends of VL. Spatial autocorrelation and spatial-temporal clustering analysis were conducted to identify the distribution and risk areas of VL transmission. RESULTS: A total of 4877 VL cases were reported in mainland China during 2004-2019, with mean annual incidence of 0.0228/100,000. VL incidence showed a decreasing trend in general during our study period (annual percentage change [APC] = -4.2564, 95% confidence interval [CI]: -8.0856 to -0.2677). Among mainly endemic provinces, VL was initially heavily epidemic in Gansu, Sichuan, and especially Xinjiang, but subsequently decreased considerably. In contrast, Shaanxi and Shanxi witnessed significantly increasing trends, especially in 2017-2019. The first-level spatial-temporal aggregation area covered two endemic provinces in northwestern China, including Gansu and Xinjiang, with the gathering time from 2004 to 2011 (relative risk [RR] = 13.91, log-likelihood ratio [LLR] = 3308.87, P < 0.001). The secondary aggregation area was detected in Shanxi province of central China, with the gathering time of 2019 (RR = 1.61, LLR = 4.88, P = 0.041). The epidemic peak of October to November disappeared in 2018-2019, leaving only one peak in March to May. CONCLUSIONS: Our findings suggest that VL is still an important endemic infectious disease in China. Epidemic trends in different provinces changed significantly and spatial-temporal aggregation areas shifted from northwestern to central China during our study period. Mitigation strategies, including large-scale screening, insecticide spraying, and health education encouraging behavioral change, in combination with other integrated approaches, are needed to decrease transmission risk in areas at risk, especially in Shanxi, Shaanxi, and Gansu provinces.

Milk casein prevents inactivation effect of black tea galloylated theaflavins on SARS-CoV-2 in vitro (preprint)

Repeated emergence of highly contagious and potentially immune-evading variant SARS-CoV-2 is posing global health and socioeconomical threats. For suppression of the spread of the virus infection among people, a procedure to inactivate virus in saliva may be useful, because saliva of infected persons is the major origin of droplets and aerosols that mediate viral transmission to nearby persons. We previously reported that SARS-CoV-2 is rapidly and remarkably inactivated by treatment in vitro with tea including green tea, roasted green tea, oolong tea and black tea. Tea catechin-derived compounds including theaflavins (TFs) with (a) galloyl moiety(ies) showed this activity. Although black tea is popularly consumed worldwide, a lot of people consume it with sugar, milk, lemon juice, and so on. But it has not been determined whether these ingredients may influence the inactivation effect of black tea against SARS-CoV-2. Moreover, it has not been revealed whether black tea is capable of inactivating variant viruses such as delta variant. Here we examined the effect of black tea on some variants in the presence or absence of sugar, milk, and lemon juice in vitro. Black tea and galloylated TFs remarkably inactivated alpha, gamma, delta and kappa variants. Intriguingly, an addition of milk but not sugar and lemon juice totally prevented black tea from inactivating alpha and delta variant viruses. The suppressive effect was also exerted by milk casein. These results suggest the possibility that intake of black tea without milk by infected persons may result in inactivation of the virus in saliva and attenuation of spread of SARS-CoV-2 to nearby persons through droplets. Clinical studies are required to investigate this possibility.

Impact of IRS: Four-years of entomological surveillance of the Indian Visceral Leishmaniases elimination programme.

BACKGROUND: In 2005, Bangladesh, India and Nepal agreed to eliminate visceral leishmaniasis (VL) as a public health problem. The approach to this was through improved case detection and treatment, and controlling transmission by the sand fly vector Phlebotomus argentipes, with indoor residual spraying (IRS) of insecticide. Initially, India applied DDT with stirrup pumps for IRS, however, this did not reduce transmission. After 2015 onwards, the pyrethroid alpha-cypermethrin was applied with compression pumps, and entomological surveillance was initiated in 2016. METHODS: Eight sentinel sites were established in the Indian states of Bihar, Jharkhand and West Bengal. IRS coverage was monitored by household survey, quality of insecticide application was measured by HPLC, presence and abundance of the VL vector was monitored by CDC light traps, insecticide resistance was measured with WHO diagnostic assays and case incidence was determined from the VL case register KAMIS. RESULTS: Complete treatment of houses with IRS increased across all sites from 57% in 2016 to 70% of houses in 2019, rising to >80% if partial house IRS coverage is included (except West Bengal). The quality of insecticide application has improved compared to previous studies, average doses of insecticide on filters papers ranged from 1.52 times the target dose of 25mg/m2 alpha-cypermethrin in 2019 to 1.67 times in 2018. Resistance to DDT has continued to increase, but the vector was not resistant to carbamates, organophosphates or pyrethroids. The annual and seasonal abundance of P. argentipes declined between 2016 to 2019 with an overall infection rate of 0.03%. This was associated with a decline in VL incidence for the blocks represented by the sentinel sites from 1.16 per 10,000 population in 2016 to 0.51 per 10,000 in 2019. CONCLUSION: Through effective case detection and management reducing the infection reservoirs for P. argentipes in the human population combined with IRS keeping P. argentipes abundance and infectivity low has reduced VL transmission. This combination of effective case management and vector control has now brought India within reach of the VL elimination targets.

Specificity of SARS-CoV-2 antibody-detection assays against S and N protein among pre-COVID-19 sera from patients with protozoan and helminth parasitic infections (preprint)

BackgroundWe aimed to assess the specificity of SARS-CoV-2 antibody detection assays among people with known tissue-borne parasitic infections. MethodsWe tested three SARS-CoV-2 antibody-detection assays (cPass SARS-CoV-2 Neutralization Antibody Detection Kit, Abbott SARS-CoV-2 IgG assay, and STANDARD Q COVID-19 IgM/IgG Combo Rapid Test) among 559 pre-COVID-19 sera. ResultsThe specificity of assays was 95-98% overall. However, lower specificity was observed among sera from patients with protozoan infections of the reticuloendothelial system, such as human African trypanosomiasis (Abbott Architect; 88% [95%CI 75-95]), visceral leishmaniasis (SD RDT IgG; 80% [95%CI 30-99]), and from patients with recent malaria from a holoendemic area of Senegal (ranging from 91% for Abbott Architect and SD RDT IgM to 98-99% for cPass and SD RDT IgG). For specimens from patients with evidence of past or present helminth infection overall, test specificity estimates were all [≥] 96%. Sera collected from patients clinically suspected of parasitic infections that tested negative for these infections yielded a specificity of 98-100%. The majority (>85%) of false-positive results were positive by only one assay. ConclusionsThe specificity of SARS-CoV-2 serological assays among sera from patients with tissue-borne parasitic infections was below the threshold required for decisions about individual patient care. Specificity is markedly increased by the use of confirmatory testing with a second assay. Finally, the SD RDT IgG proved similarly specific to laboratory-based assays and provides an option in low-resource settings when detection of anti-SARS-CoV-2 IgG is indicated.

Surveillance data for human leishmaniasis indicate the need for a sustainable action plan for its management and control, Greece, 2004 to 2018.

BackgroundThe World Health Organization (WHO) lists human leishmaniasis as a neglected tropical disease; it is not under surveillance at European level.AimWe present surveillance data for visceral (VL) and cutaneous (CL) leishmaniasis for the period 2004 to 2018 in Greece to assess their public health importance.MethodsWe extracted data from the mandatory notification system to analyse separately imported and domestic cases of VL and CL. A case was defined by clinical manifestations compatible with VL or CL and laboratory confirmation.ResultsBetween 2004 and 2018, 881 VL (862 domestic, 19 imported) and 58 CL cases (24 domestic, 34 imported) were recorded. The mean annual notification rate of domestic VL was 0.5 per 100,000 (range: 0.12-1.43/100,000) with a statistically significant increasing trend (p = 0.013). Cases were reported by all regions. The highest notification rate occurred in the age group 0-4 years (1.3/100,000). Overall 24% (164/680) of the cases were immunocompromised and their proportion increased after 2010 (p < 0.001). The mean annual notification rate of domestic CL was 0.05 per 100,000 (range: 0.01-0.19/100,000) with the highest rate in the age group 5-14 years (0.03/100,000). Cases were recorded in six of the 13 regions. Among 34 imported CL cases, 29 were foreign nationals.ConclusionVL is endemic in Greece, with an increasing trend and a considerable burden of severe disease and young children being most affected. CL is rarely reported. A sustainable action plan is needed to reduce the burden of VL and prevent local transmission of CL.