Spatiotemporal Changes in Plasmodium vivax msp142 Haplotypes in Southern Mexico: From the Control to the Pre-Elimination Phase

For 20 years, Plasmodium vivax has been the only prevalent malaria species in Mexico, and cases have declined significantly and continuously. Spatiotemporal genetic studies can be helpful for understanding parasite dynamics and developing strategies to weaken malaria transmission, thus facilitating the elimination of the parasite. The aim of the current contribution was to analyze P. vivax-infected blood samples from patients in southern Mexico during the control (1993–2007) and pre-elimination phases (2008–2011). Nucleotide and haplotype changes in the pvmsp142 fragment were evaluated over time. The majority of multiple genotype infections occurred in the 1990s, when the 198 single nucleotide sequences exhibited 57 segregating sites, 64 mutations, and 17 haplotypes. Nucleotide and genetic diversity parameters showed subtle fluctuations from across time, in contrast to the reduced haplotype diversity and the increase in the R2 index and Tajima’s D value from 2008 to 2011. The haplotype network consisted of four haplogroups, the geographical distribution of which varied slightly over time. Haplogroup-specific B-cell epitopes were predicted. Since only high-frequency and divergent haplotypes persisted, there was a contraction of the parasite population. Given that 84% of haplotypes were exclusive to Mesoamerica, P. vivax flow is likely circumscribed to this region, representing important information for parasite surveillance.

Assessment of the nlmixr R-package for population pharmacokinetic modeling: A metformin case study

Aim: nlmixr offers first-order conditional estimation with or without interaction (FOCE or FOCEi) and stochastic approximation estimation-maximisation (SAEM) to fit nonlinear mixed-effect models (NLMEM). We modelled metformin’s population pharmacokinetics with flip-flop characteristics within nlmixr framework and investigated SAEM and FOCEi’s performance with respect to bias, precision, and robustness. Method: Compartmental pharmacokinetic models were fitted. The final model was determined based on the lowest objective function value and visual inspection of goodness-of-fit plots. To examine flip-flop pharmacokinetics, k_a values of a typical concentration-time profile based on the final model were perturbed and changes in the steepness of the terminal elimination phase were evaluated. The bias and precision of parameter estimates were compared between SAEM and FOCEi using stochastic simulations and estimations. For robustness, parameters were re-estimated as the initial estimates were perturbed 100-times and resultant changes evaluated. Results: A one-compartment model with transit compartment for absorption best described the data. At low n, Stirling’s approximation of n! over-approximated plasma concentration unlike the log-gamma function. Flip-flop pharmacokinetics were evident as the steepness of the terminal elimination phase changed with k_a. Mean rRMSE for fixed-effect parameters was 0.932. When initial estimates were perturbed, FOCEi estimates of k_a and food effect on k_a appeared bimodal and were upward biased. Discussion: nlmixr is reliable for NLMEM even if flip-flop is present but caution should be exercised when using Stirling’s approximation for n! in the transit compartment model. SAEM was marginally superior to FOCEi in bias and precision, but SAEM was superior against initial estimate perturbations.

Measles in Vaccinated People: Epidemiology and Challenges in Surveillance and Diagnosis in the Post-Elimination Phase. Spain, 2014–2020

The MMR vaccination program was introduced in Spain in 1981. Consistently high vaccination coverage has led to Spain being declared free of endemic measles transmission since 2014. A few imported and import-related cases were reported during the post-elimination phase (2014 to 2020), with very low incidence: three cases per million of inhabitants a year, 70% in adults. In the post-elimination phase an increasing proportion of measles appeared in two-dose vaccinated individuals (up to 14%), posing a challenge to surveillance and laboratory investigations. Severity and clinical presentation were milder among the vaccinated. The IgM response varied and the viral load decreased, making the virus more difficult to detect. A valid set of samples (serum, urine and throat swab) is strongly recommended for accurate case classification. One third of measles in fully vaccinated people was contracted in healthcare settings, mainly in doctors and nurses, consistent with the important role of high intensity exposure in measles breakthrough cases. Surveillance protocols and laboratory algorithms should be adapted in advanced elimination settings. Reinforcing the immunity of people working in high exposure environments, such as healthcare settings, and implementing additional infection control measures, such as masking and social distancing, are becoming crucial for the global aim of measles eradication.

Pharmacokinetic, Clinical, and Myeloid Marker Responses to Acepromazine Sedation in Arabian Camels

Sedatives and tranquilizers are important in the control of excited camels during camel transport. This study was conducted to investigate the clinical sedation of camels with acepromazine and its correlation with pharmacokinetics and pharmacodynamics. The sedation score, heart rate, respiration, body temperature, and pharmacokinetics were monitored before and after acepromazine injection, and myeloid marker expression was analyzed using membrane immunofluorescence and flow cytometry. The distribution (t1/2α) and elimination (t1/2β) half-lives were 0.1 and 9.4 h, respectively. The volume of distribution at steady state (Vss) was 20.01 L/kg, and the mean residence time (MRT) was 12.25 h. Sedation started rapidly within 10 min followed by persistent low-medium sedation for 2 h with an average sedation score of 1.2 ± 0.61, which might be associated with a slow elimination phase and prolonged MRT. Compared to horses, camels showed a lower clearance rate, higher volume of distribution, and higher elimination half-life. Slight changes in body temperature and heart and respiratory rate, as well as a lower hematocrit and changes in blood cell composition, suggest the careful application of acepromazine in animals with abnormal blood parameters or poor vital conditions.

KAJIAN EPIDEMIOLOGI P. KNOWLESI DI PROVINSI ACEH TAHUN 2018-2019

Kementerian Kesehatan Republik Indonesia telah menetapkan Indonesia akan mencapai fase eliminasi malaria pada tahun 2030. Tapi, periode tahun 2018-2019 di Provinsi Aceh dilaporkan masih adanya temuan 19 kasus positif infeksi P. knowlesi. Penelitian ini bertujuan melakukan analisis faktor risiko terhadap kasus infeksi P. knowlesi di Provinsi Aceh periode tahun 2018 s/d 2019. Penelitian observasional dengan desain penelitian kasus kontrol, dilakukan di 4 wilayah kabupaten/ kota yang menjadi lokasi penemuan kasus infeksi P. knowlesi di Provinsi Aceh. Populasinya seluruh masyarakat dari 4 kabupaten/kota yang menjadi lokasi penelitian yang potensial tinggal dekat hutan. Sampel penelitian sebanyak 74 orang (16 kasus: 58 kontrol). Analisa data dilakukan dengan menggunakan analisis regresi logistik. Hasil uji analisis bivariat didapatkan ada hubungan antara kebiasaan keluar malam hari (p=0,001) dengan kejadian infeksi P. knowlesi. Selanjutnya tidak ada hubungan antara keberadaan macaca, lingkungan hutan, obat anti nyamuk, kelambu, dan kawat kasa dengan kejadian infeksi P. knowlesi. Hasil analisis multivariat untuk keseluruhan variabel penelitian diketahui bahwa kebiasaan keluar malam hari merupakan prediktor paling dominan terhadap kejadian infeksi P. knowlesi ((OR=11,25) (95% CI: 3,01 – 42,08) p Value =0,000). Hasil penelitian secara umum menyimpulkan bahwa kebiasaan keluar malam hari secara signifikan memiliki pengaruh terhadap kejadian infeksi P. knowlesi di provinsi Aceh. The Ministry of Health of the Republic of Indonesia has determined that Indonesia will reach the elimination phase of malaria in 2030. But, in period 2018 - 2019 Aceh Province is reported to have found 19 positive cases of P. knowlesi infection. This research aims to dorisk factor analysis of cases P. knowlesi infection in Aceh Province for the period 2018 to 2019. This observational study with a case control research design was conducted in 4 districts/ cities which were the location for finding cases of P. knowlesi infection in Aceh Province. Its populationAll people from the 4 districts / cities that are potential research locations live near the forest. The research sample was 74 people (16 cases: 58 control). Data analysis was performed using logistic regression analysis. The results of the bivariate analysis test showed that there was a relationship between the habit of going out at night (p=0.001) with the incidence of P. knowlesi infection. Furthermore, there is no relationship between the presence of macaca, forest environment, mosquito repellent, mosquito nets, and gauze with the incidence of P. knowlesi infection. The results of multivariate analysis for all research variables showed that nighttime out habits were the most dominant predictor of P. knowlesi infection ((OR = 11.25) (95% CI: 3.01 - 42.08) p value = 0.000). The results of the study generally concluded that the habit of going out at night had a significant effect on the incidence of P. knowlesi infection in Aceh province.

Simplified Iohexol-Based Method for Measurement of Glomerular Filtration Rate in Goats and Pigs

The preclinical evaluation of novel therapies for chronic kidney disease requires a simple method for the assessment of kidney function in a uremic large animal model. An intravenous bolus of iohexol was administered to goats (13 measurements in n = 3 goats) and pigs (23 measurements in n = 5 pigs) before and after induction of kidney failure, followed by frequent blood sampling up to 1440 min. Plasma clearance (CL) was estimated by a nonlinear mixed-effects model (CLNLME) and by a one-compartmental pharmacokinetic disposition model using iohexol plasma concentrations during the terminal elimination phase (CL1CMT). A simple method (CLSM) for the calculation of plasma clearance was developed based on the most appropriate relationship between CLNLME and CL1CMT. CLSM and CLNLME showed good agreement (CLNLME/CLSM ratio: 1.00 ± 0.07; bias: 0.03 ± 1.64 mL/min; precision CLSM and CLNLME: 80.9% and 80.7%, respectively; the percentage of CLSM estimates falling within ±30% (P30) or ±10% (P10) of CLNLME: 53% and 12%, respectively). For mGFRNLME vs. mGFRSM, bias was −0.25 ± 2.24 and precision was 49.2% and 53.6%, respectively, P30 and P10 for mGFR based on CLSM were 71% and 24%, respectively. A simple method for measurement of GFR in healthy and uremic goats and pigs was successfully developed, which eliminates the need for continuous infusion of an exogenous marker, urine collection and frequent blood sampling.

Dynamic changes in genetic diversity, drug resistance mutations, and treatment outcomes of falciparum malaria from the low-transmission to the pre-elimination phase on the isolated islands of São Tomé and Príncipe

Background With effective vector control and case management, substantial progress has been made in the elimination of malaria on the islands of São Tomé and Príncipe (STP). During the critical period from the low-transmission to the pre-elimination phase, this study tracked the dynamic changes in the genetic diversity in Plasmodium falciparum, the distribution of antimalarial drug-resistance genes, and the treatment outcomes in patients to provide insights for the prevention of rebounded malaria in STP. Methods Dried blood spots (DBSs) and case follow-up data were collected from malaria patients who had visited the Central Hospital between 2010 and 2016. Genomic DNA of P. falciparum was extracted from DBSs. The polymorphic regions on the genes for merozoite surface proteins 1 and 2 (msp1 and msp2) were amplified in 118 pre-treatment samples to identify the genetic diversity of the infected parasites. Anti-malarial drug resistance mutations in the multi-drug resistance (pfmdr1), chloroquine resistance transporter (pfcrt), and kelch 13 (pfK13) genes were genotyped using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and DNA sequencing in 111 samples. Treatment outcomes were categorized based on the parasitological results from microscopy during the 28-day follow-up after treatment. Factors related to malaria recurrence were characterized by logistic regression models using case follow-up data (total number = 7,482). Results The circulating parasite strains in STP showed significant changes at the recent peak incidence in 2012, during which the prevalent allelic type in MSP1 changed from K1 to MAD20, and that in MSP2 changed from 3D7/IC to FC27. Genotyping results for antimalarial drug-resistance markers showed that the dominant alleles of pfmdr1 86 + 184 + 1246-pfcrt 76 were YFD-T (51.4%). Logistic regression models showed that significant factors related to parasitological failure after treatment were age (protective factor, OR = 0.97–0.98), log10-transformed parasite density (OR = 1.07–1.44), and treatment (quinine vs. artemisinin-based combination therapy, OR = 1.91–1.96). Overall, younger patients, those with higher parasitemia levels at enrollment, and those treated with quinine had a higher risk of recurrence during follow-up. Conclusions Although malaria treatment efficacy remained acceptable in STP, this study showed temporal changes in the dominant strains and the development of drug resistance mutations in the local parasite population. Therapeutic efficacy should be carefully monitored to adequately adjust the policy in the future.

Spatial patterns of Plasmodium vivax transmission explored by multivariate auto-regressive state-space modelling - A case study in Baoshan Prefecture in southern China

The transition from the control phase to elimination of malaria in China through the national malaria elimination programme has focussed attention on the need for improvement of the surveillance- response systems. It is now understood that routine passive surveillance is inadequate in the parasite elimination phase that requires supplementation by active surveillance in foci where cluster cases have occurred. This study aims to explore the spatial clusters and temporal trends of malaria cases by the multivariate auto-regressive state-space model (MARSS) along the border to Myanmar in southern China. Data for indigenous cases spanning the period from 2007 to 2010 were extracted from the China’s Infectious Diseases Information Reporting Management System (IDIRMS). The best MARSS model indicated that malaria transmission in the study area during 36 months could be grouped into three clusters. The estimation of malaria transmission patterns showed a downward trend across all clusters. The proposed methodology used in this study offers a simple and rapid, yet effective way to categorize patterns of foci which provide assistance for active monitoring of malaria in the elimination phase.

Lymphatic filariasis, infection status in Culex quinquefasciatus and Anopheles species after six rounds of mass drug administration in Masasi District, Tanzania

Background Lymphatic filariasis (LF) elimination program in Tanzania started in 2000 in response to the Global program for the elimination of LF by 2020. Evidence shows a persistent LF transmission despite more than a decade of mass drug administration (MDA). It is advocated that, regular monitoring should be conducted in endemic areas to evaluate the progress towards elimination and detect resurgence of the disease timely. This study was therefore designed to assess the status of Wuchereria bancrofti infection in Culex quinqefasciatus and Anopheles species after six rounds of MDA in Masasi District, South Eastern Tanzania. Methods Mosquitoes were collected between June and July 2019 using Center for Diseases Control (CDC) light traps and gravid traps for indoor and outdoor respectively. The collected mosquitoes were morphologically identified into respective species. Dissections and PCR were carried out to detect W. bancrofti infection. Questionnaire survey and checklist were used to assess vector control interventions and household environment respectively. A Poisson regression model was run to determine the effects of household environment on filarial vector density. Results Overall, 12 452 mosquitoes were collected of which 10 545 (84.7%) were filarial vectors. Of these, Anopheles gambiae complex, An. funestus group and Cx. quinquefasciatus accounted for 0.1%, 0.7% and 99.2% respectively. A total of 365 pools of Cx. quinquefasciatus (each with 20 mosquitoes) and 46 individual samples of Anopheles species were analyzed by PCR. For Cx. quinquefasciatus pools, 33 were positive for W. bancrofti, giving an infection rate of 0.5%, while the 46 samples of Anopheles species were all negative. All 1859 dissected mosquitoes analyzed by microscopy were also negative. Households with modern latrines had less mosquitoes than those with pit latrines [odds ratio (OR) = 0.407, P < 0.05]. Houses with unscreened windows had more mosquitoes as compared to those with screened windows (OR = 2.125, P < 0.05). More than 80% of the participants own bednets while 16.5% had no protection. Conclusions LF low transmission is still ongoing in Masasi District after six rounds of MDA and vector control interventions. The findings also suggest that molecular tools may be essential for xenomonitoring LF transmission during elimination phase.