Aptamer-Based Technologies for Parasite Detection.

Centuries of scientific breakthroughs have brought us closer to understanding and managing the spread of parasitic diseases. Despite ongoing technological advancements in the detection, treatment, and control of parasitic illnesses, their effects on animal and human health remain a major concern worldwide. Aptamers are single-stranded oligonucleotides whose unique three-dimensional structures enable them to interact with high specificity and affinity to a wide range of targets. In recent decades, aptamers have emerged as attractive alternatives to antibodies as therapeutic and diagnostic agents. Due to their superior stability, reusability, and modifiability, aptamers have proven to be effective bioreceptors for the detection of toxins, contaminants, biomarkers, whole cells, pathogens, and others. As such, they have been integrated into a variety of electrochemical, fluorescence, and optical biosensors to effectively detect whole parasites and their proteins. This review offers a summary of the various types of parasite-specific aptamer-based biosensors, their general mechanisms and their performance.

[Bacterial zoonoses of public health importance in Germany-incidence, distribution, and modes of transmission]. / Bakterielle Zoonosen mit Bedeutung für den öffentlichen Gesundheitsschutz in Deutschland ­ Vorkommen, Verbreitung und Übertragungswege.

Bacterial zoonotic pathogens are often the cause of diseases, sometimes with severe outcomes. They are mutually transferable between animals (both wild and domestic) and humans. The transmission paths are very variable and include oral intake via food, respiratory infection via droplets and aerosols, or infections via vectors such as tick bites or rodent contact. Furthermore, the emergence and spread of antibiotic-resistant bacterial pathogens is of paramount public health concern.The likelihood of further spread is influenced by various factors. These include the increase in international trade, the endangerment of animal habitats, and the increasingly closer contact between humans and wild animals. Additionally, changes in livestock and climate change may also contribute. Therefore, research into zoonoses serves to protect human and animal health and is of particular social, political, and economic importance.The aim of this review article is to present the range of infectious diseases caused by bacterial zoonotic pathogens in order to provide a better understanding of the important work in public health services, animal health services, and food safety control. The different transmission routes, epidemic potentials, and epidemiological measures of the exemplary selected diseases show the challenges for the public health system to monitor and control the spread of these bacterial pathogens in order to protect the population from disease.

Risk factors for Leptospira seropositivity in rural Northern Germany, 2019.

We investigated seroprevalence and factors associated with Leptospira spp. infections in humans in rural Northern Germany. Sera of 450 participants were tested for leptospira-reactive IgG antibodies by two enzyme-linked immunosorbent assays (ELISA). A narrow (specific) and a broad (sensitive) case definition were applied and results compared in the analysis. Personal data were collected via questionnaire and associations with the serostatus were investigated by multivariable logistic regression. The seroprevalence estimates were 1.6% (95%-confidence interval (CI) = 0.63-3.2) under the narrow and 4.2% (95%-CI = 2.6-6.5%) under the broad case definition. Few (14%) participants knew about the pathogen. No seropositive participant recalled a prior leptospirosis diagnosis. Spending more than two hours a week in the forest was significantly associated with anti-leptospira IgG in both models (broad case definition: adjusted odds ratio (aOR) = 2.8, 95%-CI = 1.2-9.1; narrow case definition: aOR = 11.1, 95%-CI = 1.3-97.1). Regular cleaning of storage rooms was negatively associated in the broad (aOR = 0.17, 95%-CI = 0.03-0.98) and touching a dead rodent in the past 10 years in the narrow case definition model (aOR = 0.23, 95%-CI = 0.05-1.04). Our findings support risk factors identified in previous investigations. To counter the low awareness for the pathogen, we recommend that health authorities communicate risks and preventive measures to the public by using target-group specific channels.

Prevalence of Alaria alata mesocercariae in wild boars from Brandenburg, Germany.

Since 2002, Alaria (A.) alata mesocercariae (AM) have been found during routine Trichinella inspection of wild boars in many European countries. To date, human infection with AM through consumption of undercooked or raw AM infested wild boar meat cannot be excluded. In Germany, data on the parasite's prevalence in wild boars are scarce. To better understand temporal and spatial fluctuations of this parasite, this study investigated the prevalence of AM in wild boars in the German federal state of Brandenburg during three hunting seasons from 2017 to 2020. In total, 28.3% (100/354, 95% CI: 23.3-33.3%) of all wild boars sampled in eight counties of Brandenburg were tested positive for AM by Alaria alata mesocercariae migration technique (AMT). AM were detected in wild boars from seven different counties. Samples from one county (Havelland) tested completely negative for AM (0/16). Prevalences of the seven AM positive counties of Brandenburg ranged from 11.5 (3/26, 95% CI: 2.5-30.1%) in Märkisch-Oderland to 64.1% (25/39, 95% CI: 47.2-78.8%) in Uckermark. An association between sex and A. alata positivity could not be determined. A statistically significant increase in frequency of older AM positive wild boars was observed (p = 0.001). For a nationwide assessment of the prevalence of A. alata in wild boars and the risk for consumers of ingesting viable AM by consumption of raw or undercooked AM infested wild boar meat, further long-term studies in different regions of Germany are needed.

Species identification of Trichinella originated from various host and different geographical location by MALDI-TOF.

The foodborne zoonotic nematode Trichinella spp. can cause human trichinellosis when raw or undercooked contaminated meat is ingested. To date, twelve Trichinella species/genotypes have been described. According to EU regulation any Trichinella larvae detected during mandatory routine examinations need to be identified at a species level by a competent laboratory. Currently, Trichinella species identification is performed using molecular biology tools such as multiplex PCR, PCR-sequencing or PCR-RFLP. These techniques require high level of skills for good interpretation of the results. Due to its rapidness and ease of use a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) protocol was previously developed for the identification of Trichinella species. Using this method, spectra from different Trichinella species and strains were acquired allowing to generate new Main Spectra (MSP). Finally a new MSP database from Trichinella spp. Samples of different countries (France, Germany and Poland), including field samples, was generated. Comparing the different main spectra, Trichinella worms were identified at the species level and differences in the genetic diversities within the different species are discussed. In conclusion, using the previously described method on field samples is a reliable, rapid, easy-to-use and cheap tool for Trichinella species identification. The new Trichinella database could be incremented with new samples. It constitutes a tool, which could be used as an alternative method to replace the actual molecular methods for Trichinella species identification.

A two-step morphology-PCR strategy for the identification of nematode larvae recovered from muscles after artificial digestion at meat inspection.

To ensure that meat from livestock and game is safe for human consumption, European legislation lays down rules for mandatory testing. Helminth larvae are a category of zoonotic foodborne pathogens that can contaminate meat. Among helminths, the only zoonotic nematode regulated in Europe regarding meat inspection is Trichinella spp.. It is precisely during Trichinella testing that other potentially zoonotic larvae can be found. Due to current lack of tools, their identification is often very complicated. Nematode larvae other than Trichinella, recovered from artificial digestions of pig and wild boar muscles from France and Germany, were subjected to a newly developed two-step identification scheme, which includes both morphological examination and molecular assays. The first step is a general orientation towards a broad taxonomic group; the second step consists of targeted identification based on the results of first step. Different parasites were identified, some of which were not zoonotic such as Metastrongylus spp. and Angiostrongylus vasorum, but others are known to be zoonotic such as Toxocara cati, Ascaris suum, and Uncinaria stenocephala. The strategy is efficient for the identification of nematode larvae recovered from muscles but could also be applied for larvae from other sources.

[Leptospirosis in Germany: current knowledge on pathogen species, reservoir hosts, and disease in humans and animals]. / Leptospirose in Deutschland: Aktuelle Erkenntnisse zu Erregerspezies, Reservoirwirten und Erkrankungen bei Mensch und Tier.

Leptospirosis is a zoonotic disease with a wide spectrum of clinical symptoms in humans and animals, ranging from subclinical infections to severe signs of multiorgan dysfunction. In Germany, laboratory confirmation of acute human infection is notifiable based on the Protection Against Infection Act. Disease or occurrence of the pathogen in pigs and sheep must be reported according to the regulation on reportable animal diseases. Transmission occurs via direct and indirect contact with the urine of infected animals, with rodents acting as the main reservoir. With an average annual incidence of 0.1 notified cases per 100,000 inhabitants, leptospirosis is a rare disease in Germany.This review article presents the current knowledge on leptospirosis in Germany in the framework of the project "Improving public health through a better understanding of the epidemiology of rodent-transmitted diseases" (RoBoPub) funded by the Ministry of Education and Research. In a One-Health approach, information about clinical manifestation, available prevalence data in humans and animals, knowledge of pathogen distribution, host association, mode of transmission, and survival in the environment is summarized. Preliminary findings on the influence of fluctuations in rodent populations on the occurrence of leptospirosis are also discussed. The aim of the article is to increase the awareness of this currently neglected disease in Germany.In future, higher temperatures and more frequent heavy rainfalls, which could occur due to climate change, should be taken into account.

Outbreak of trichinellosis due to wild boar meat and evaluation of the effectiveness of post exposure prophylaxis, Germany, 2013.

BACKGROUND: Food safety authorities discovered that wild boar meat products contaminated with Trichinella spiralis had entered the food chain in Germany in March 2013. Public health authorities issued guidelines for health professionals including post-exposure prophylaxis (PEP) using mebendazole and advised the public to seek medical advice if exposed. Our objective was to identify factors associated with the development of trichinellosis and to evaluate post exposure prophylaxis. METHODS: Persons who reported to local public health departments as exposed were interviewed concerning exposure, symptoms, and medication. Serum samples were tested by an in-house Trichinella-specific enzyme-linked innunosorbent assay. Cases were defined as persons presenting with myalgia and/or periorbital edema and Trichinella-specific immunoglobulin M and immunoglobulin G antibodies after exposure to implicated products. RESULTS: Of 101 persons interviewed, 71 were exposed and serologically tested. Antibodies were detected in 21/71 (30%) and 14/71 (20%) met the case definition. Attack rates were positively correlated to the amount of implicated product consumed. Among n = 37 persons who received anthelmintics as PEP, 6 persons developed trichinellosis. These cases exclusively occurred among persons starting PEP 6 days or later post-exposure. Exposure to implicated products and delaying PEP were also significantly associated with developing trichinellosis (P < .01) in a multivariable analysis. CONCLUSIONS: Concerted efforts by food safety and public health authorities lead to timely outbreak control and facilitated the provision of early PEP. PEP appears to be effective in preventing trichinellosis when given early, preferably within 6 days. We therefore recommend initiating PEP without delay in similar settings and encourage public health professionals to fast-track this intervention.

Estimation of the sensitivity and specificity of a Leptospira spp. in-house ELISA through Bayesian modelling.

The microscopic agglutination test (MAT) is still considered the gold standard for the diagnosis of leptospirosis, although studies have shown that the test is an imperfect gold standard for clinical samples and unsuitable for epidemiological studies. Here, test characteristics of an in-house ELISA were identified for both subclinical and clinical populations by Bayesian latent class models. A conditional dependence model for two diagnostic tests and two populations was adapted to analyse a clinical and a subclinical scenario, respectively. These Bayesian models were used to estimate the sensitivity and specificity of the in-house ELISA and the MAT as well as the prevalences. The Bayesian estimates of the in-house ELISA were: clinical sensitivity=83.0%, clinical specificity=98.5%, subclinical sensitivity=85.7% and subclinical specificity=99.1%. In contrast, the estimates of the MAT were: clinical sensitivity=65.6%, clinical specificity=97.7%, subclinical sensitivity 54.9% and subclinical specificity=97.3%. The results show the suitability of the in-house ELISA for both clinical investigations and epidemiological studies in mildly endemic areas.

Severe leptospirosis complicated by Epstein-Barr Virus reactivation.

INTRODUCTION: Weil's disease is a severe, potentially fatal illness following Leptospira interrogans infection. The reported case of a patient suffering from acute renal failure, jaundice, thrombocytopenia, rhabdomyolysis and encephalitis syndrome highlights the clinical challenge in reference to Weil syndrome complicated by Epstein-Barr Virus (EBV) reactivation. MATERIALS AND METHODS: The diagnosis of leptospirosis was performed using four different diagnostic methods. Sera were analyzed with an in-house IgM and IgG enzyme-linked immunosorbent assay (ELISA) and indirect haemagglutination assay (IHA). Microscopic agglutination test (MAT) was done using 17 reference strains comprising 14 serogroups and 17 serovars. Polyvalent EBV-IgG analysis, EBV-IgG/IgM/IgA western blot analysis as well as quantitative EBV polymerase chain reaction (PCR) were performed. RESULTS: Leptospira IHA showed an initial titer of 1:640 (cut-off 1:320), leptospiral IgG was negative, but IgM was positive. MAT was negative at that time for all 17 strains analyzed. One week later, leptospirosis IHA titer increased to 1:20,480. Leptospiral IgG was now positive, -IgM remained positive and urine was tested negative for leptospiral DNA. The MAT showed positive results for L. interrogans serovar Bataviae, serovar Copenhageni, serovar Pyrogenes and L. borgpetersenii serovar Serjoe. During follow-up examinations, both the leptospiral IgM and IgG remained positive and MAT showed positive results for L. interrogans of different serovars. EBV IgA immunoblot taken at admission was positive for VCA-p18, quantitative EBV-PCR showed an EBV viral load of 2.8E3 copies/ml indicating acute EBV-reactivation. CONCLUSION: Leptospirosis represents a neglected and re-emerging disease which is difficult to diagnose since Leptospira-PCR from whole blood or urine is frequently negative in the case of early empiric antibiotic treatment. EBV-reactivation might represent a severe complication in Weil's disease which potentially aggravates clinical manifestations of leptospirosis including hepatitis, nephritis, and rhabdomyolysis. Thus, there might be a need for peripheral blood EBV-PCR and EBV blotting in patients suffering from complicated Weil syndrome, also in terms of the choice of antibiotic treatment.

Molecular and serological diagnosis of multiple bacterial zoonoses in febrile outpatients in Garissa County, north-eastern Kenya.

Bacterial zoonoses are diseases caused by bacterial pathogens that can be naturally transmitted between humans and vertebrate animals. They are important causes of non-malarial fevers in Kenya, yet their epidemiology remains unclear. We investigated brucellosis, Q-fever and leptospirosis in the venous blood of 216 malaria-negative febrile patients recruited in two health centres (98 from Ijara and 118 from Sangailu health centres) in Garissa County in north-eastern Kenya. We determined exposure to the three zoonoses using serological (Rose Bengal test for Brucella spp., ELISA for C. burnetti and microscopic agglutination test for Leptospira spp.) and real-time PCR testing and identified risk factors for exposure. We also used non-targeted metagenomic sequencing on nine selected patients to assess the presence of other possible bacterial causes of non-malarial fevers. Considerable PCR positivity was found for Brucella (19.4%, 95% confidence intervals [CI] 14.2-25.5) and Leptospira spp. (1.7%, 95% CI 0.4-4.9), and high endpoint titres were observed against leptospiral serovar Grippotyphosa from the serological testing. Patients aged 5-17 years old had 4.02 (95% CI 1.18-13.70, p-value = 0.03) and 2.42 (95% CI 1.09-5.34, p-value = 0.03) times higher odds of infection with Brucella spp. and Coxiella burnetii than those of ages 35-80. Additionally, patients who sourced water from dams/springs, and other sources (protected wells, boreholes, bottled water, and water pans) had 2.39 (95% CI 1.22-4.68, p-value = 0.01) and 2.24 (1.15-4.35, p-value = 0.02) times higher odds of exposure to C. burnetii than those who used unprotected wells. Streptococcus and Moraxella spp. were determined using metagenomic sequencing. Brucellosis, leptospirosis, Streptococcus and Moraxella infections are potentially important causes of non-malarial fevers in Garissa. This knowledge can guide routine diagnosis, thus helping lower the disease burden and ensure better health outcomes, especially in younger populations.

Anti-Leptospira Seroprevalence and Associated Risk Factors among Forestry Workers in Lower Saxony, North-West Germany.

As leptospirosis is re-emerging, a seroprevalence study was conducted, assessing the prevalence of anti-Leptospira IgG antibodies and infection-associated risk factors among forestry workers (FWs) in Lower Saxony, Germany, to develop targeted public health measures. Sera of 877 FWs, sampled in 2016, were tested for anti-Leptospira seropositivity by commercial IgG-ELISA. Data on demographics and Leptospira-specific exposures, knowledge, sources of information, and preventive measures were collected by standardized, self-administered questionnaire. A subset of 244 sera was retested via in-house IgG-ELISA. Risk factors were assessed from the subset using multivariable logistic regression analysis. The commercial IgG-ELISA revealed a seroprevalence of 4.8% (95% confidence interval CI95 = 3.5-6.4). Of the 601 FWs who completed the questionnaire, 67.9% had been informed about leptospirosis and Leptospira spp., mainly by employers (55.2%) and peers (38.9%). Positive associations with seropositivity were observed for canoeing (adjusted odds ratio (aOR) = 2.35, p = 0.044), touching rodents (aOR = 2.4, p = 0.021), and living close to beech trees (aOR = 2.18, p = 0.075). Frequently cleaning animal stables was negatively associated (aOR = 0.20, p = 0.002). The unexpected positive association with wearing gloves when handling plants and soil (aOR = 2.16, p = 0.011) needs further discussion. Overall, seroprevalence was in the range of other studies in Germany. The identified factors will be used to develop targeted information reaching out to at-risk groups tapping various communication channels.

Cross-sectional serosurvey of Leptospira species among slaughter pigs, goats, and sheep in Uganda.

INTRODUCTION: Leptospira are a group of bacteria, including pathogenic types that cause leptospirosis. In Uganda, Leptospira exposure has been reported in humans, with domesticated animals being speculated as the source. However, comparable evidence of Leptospira prevalence and circulating serovars/serogroups in animals is only documented for cattle, and dogs. Our study determined Leptospira seroprevalence, associated risk factors and serogroups circulating among slaughtered pigs, goats, and sheep in Uganda. METHODS: During an 11-month cross-sectional survey in selected slaughter facilities in three regions of Uganda, we collected blood from 926 pigs, 347 goats, and 116 sheep. The age, sex, breed, and origin of each sampled animal were noted. The samples were tested for anti-Leptospira antibodies using the microscopic agglutination test, based on a panel of 12 serovars belonging to 12 serogroups. RESULTS: Leptospira seroprevalence was 26.67% (247/926, 95%CI 23.92-29.61) among pigs, and 21.81% (101/463, 95%CI 18.29-25.80) in goats and sheep (small ruminants). L. interrogans Australis and L. kirschneri Grippotyphosa were the commonest serovars among pigs, as was L. borgpetersenii Tarassovi in small ruminants. Pigs sourced from the Eastern (Odds Ratio [OR] = 2.82, 95%CI 1.84-4.30) and Northern (OR = 3.56, 95%CI 2.52-5.02) regions were more likely to be seropositive, compared to those from the Central region. For small ruminants, being female (OR 2.74, 95% CI 1.69-4.57) and adult (OR 4.47, 95% CI 1.57-18.80) was significantly more associated with Leptospira seropositivity. Conclusion/significance: Detection of a moderate seroprevalence, and several Leptospira serogroups among pigs, sheep, and goats from all regions of Uganda, supports existing reports in cattle and dogs, and implies widespread Leptospira exposure in domestic animals in Uganda. These findings may inform future programs for the control of leptospirosis in livestock in Uganda.

Genetic identification of Trichinella species found in wild carnivores from the territory of Kazakhstan.

Trichinellosis, also called trichinosis, is a foodborne parasitic disease caused by eating raw or undercooked meat from animals infected with Trichinella spp. larvae and affects both animals and humans. Although on the territory of Kazakhstan, the species characteristics and prevalence of this helminth were studied back in the 90s, the data have not been updated since then. Given the above, our study was aimed at identifying Trichinella spp. using parasitological and molecular genetics methods. In our work, we studied 160 samples of muscle tissue of wild animals living in the natural zones of steppes and semi-deserts. Of the animals examined, 32 were positive for Trichinella spp., including 1 lynx (Lynx lynx), 17 wolves (Canis lupus), 11 foxes (Vulpes vulpes), 1 jackal (Canis aureus) and 2 corsac foxes (Vulpes corsac). Helminths were extracted using the digestion method. DNA was extracted using a Gene Jet commercial kit (Thermo Fisher Scientific, United Kingdom). For species identification a multiplex PCR, amplification of ESV, ITS1, and ITS2 genes regions was performed. After that, uniplex PCR was performed on the 5S rDNA and ITS1 genes region for sequencing analysis. The resulting sequences were subsequently used to construct a phylogenetic tree and the studied samples were identified as Trichinella nativa and Trichinella britovi. Thus, we can conclude that there is a circulation of two species of Trichinella in Kazakhstan, highlighting that constant control and monitoring of wild animals are necessary to prevent transmission and protect the health of people.

Investigation and response to a large outbreak of leptospirosis in field workers in Lower Saxony, Germany.

Between June and August 2014, 45 cases of leptospirosis were notified among workers on two strawberry farms in North-West Germany. We describe the characteristics of the outbreak and the actions taken to prevent further cases. The activities of the local, federal and national public health and veterinary authorities included collection of case data, laboratory testing of human specimens and of small mammals trapped on the fields, investigation of weather data, as well as information provided to farmers, field workers, physicians and to the authorities in Poland and Romania. Of the 45 identified cases (median age 22, 60% male), 47% were hospitalized. Characteristic symptoms were fever ≥38.5°C, generalized muscle pain and an increase in renal or liver enzymes. Thirteen cases were laboratory confirmed by serological and/or molecular methods. ELISA tests for Leptospira IgG and IgM-antibodies were positive in those samples taken >5 days after hospitalization. The probable causative agent was identified as Leptospira kirschneri serovar Grippotyphosa. Leptospira-specific DNA was found in kidneys of 67% of 64 trapped small mammals and was further identified as Leptospira kirschneri multi locus sequence type 110. During the estimated time period of human infections, the affected region faced warm weather with heavy rainfalls. The results of this investigation are in accordance with the theory of a chain of infection from mice to field workers during warm and humid weather. In 2015, a campaign was initiated to inform physicians, farmers and workers to enhance prevention measures, such as the use of personal protective equipment and early consultation of physicians in case of illness. Since then, no further outbreak occurred.

First detection of Trichinella spiralis in raccoon (Procyon lotor) in Germany.

Trichinella spp. are foodborne parasites that can cause severe and potentially fatal disease in humans. Infections occur through consumption of meat containing the infectious stage (L1). In Germany the domestic cycle has been eradicated. In wild animals sporadic occurrence is observed in species such as wild boar, red foxes and raccoon dogs. The omnivore raccoon which is an invasive species in Europe is known as a potential host but has not been studied intensely regarding this parasite in Germany until now, thus resulting in a lack of knowledge about its role in the sylvatic cycle. Raccoons from the urban area of Leipzig were investigated for several pathogens including Trichinella spp. in a cooperative project. Muscle samples of 88 individuals were examined using the artificial digestion method (ADM). One animal was found positive, which is the first detection of this parasite in a raccoon in Germany.

Longitudinal Study of Selected Bacterial Zoonoses in Small Ruminants in Tana River County, Kenya.

Brucellosis, Q fever, and leptospirosis are priority zoonoses worldwide, yet their epidemiology is understudied, and studies investigating multiple pathogens are scarce. Therefore, we selected 316 small ruminants in irrigated, pastoral, and riverine settings in Tana River County and conducted repeated sampling for animals that were initially seronegative between September 2014 and June 2015. We carried out serological and polymerase chain reaction tests and determined risk factors for exposure. The survey-weighted serological incidence rates were 1.8 (95% confidence intervals [CI]: 1.3-2.5) and 1.3 (95% CI: 0.7-2.3) cases per 100 animal-months at risk for Leptospira spp. and C. burnetii, respectively. We observed no seroconversions for Brucella spp. Animals from the irrigated setting had 6.83 (95% CI: 2.58-18.06, p-value = 0.01) higher odds of seropositivity to C. burnetii than those from riverine settings. Considerable co-exposure of animals to more than one zoonosis was also observed, with animals exposed to one zoonosis generally having 2.5 times higher odds of exposure to a second zoonosis. The higher incidence of C. burnetii and Leptospira spp. infections, which are understudied zoonoses in Kenya compared to Brucella spp., demonstrate the need for systematic prioritization of animal diseases to enable the appropriate allocation of resources.

Cocirculation of Leptospira spp. and multiple orthohantaviruses in rodents, Lithuania, Northern Europe.

In Europe, zoonotic Leptospira spp. and orthohantaviruses are mainly associated with specific rodent hosts. These pathogens cause febrile human diseases with similar symptoms and disease progression. In Lithuania, the presence of Dobrava-Belgrade orthohantavirus (DOBV), Tula orthohantavirus (TULV) and Leptospira spp. in rodent reservoirs is still unknown, and Puumala orthohantavirus (PUUV) was detected in bank voles (Clethrionomys glareolus) at only one site. Therefore, we collected and screened 1617 rodents and insectivores from Lithuania for zoonotic (re-)emerging Leptospira and orthohantaviruses. We detected Leptospira DNA in six rodent species, namely striped field mouse (Apodemus agrarius), yellow-necked mouse (Apodemus flavicollis), bank vole, common vole (Microtus arvalis), field vole (Microtus agrestis) and root vole (Microtus oeconomus). Leptospira DNA was detected with an overall mean prevalence of 4.4% (range 3.7%-7.9% per rodent species). We detected DOBV RNA in 5.6% of the striped field mice, PUUV RNA in 1% of bank voles and TULV RNA in 4.6% of common voles, but no Leptospira DNA in shrews and no hantavirus-Leptospira coinfections in rodents. Based on the complete coding sequences of the three genome segments, two distant DOBV phylogenetic lineages in striped field mice, one PUUV strain in bank voles and two TULV strains in common voles were identified. The Leptospira prevalence for striped field mice and yellow-necked mice indicated a significant negative effect of the distance to water points. The detection of (re-)emerging human pathogenic Leptospira and three orthohantaviruses in rodent reservoirs in Lithuania calls for increased awareness of public health institutions and allows the improvement of molecular diagnostics for pathogen identification.

A systematic review and meta-analysis of the aetiological agents of non-malarial febrile illnesses in Africa.

BACKGROUND: The awareness of non-malarial febrile illnesses (NMFIs) has been on the rise over the last decades. Therefore, we undertook a systematic literature review and meta-analysis of causative agents of non-malarial fevers on the African continent. METHODOLOGY: We searched for literature in African Journals Online, EMBASE, PubMed, Scopus, and Web of Science databases to identify aetiologic agents that had been reported and to determine summary estimates of the proportional morbidity rates (PMr) associated with these pathogens among fever patients. FINDINGS: A total of 133 studies comprising 391,835 patients from 25 of the 54 African countries were eligible. A wide array of aetiologic agents were described with considerable regional differences among the leading agents. Overall, bacterial pathogens tested from blood samples accounted for the largest proportion. The summary estimates from the meta-analysis were low for most of the agents. This may have resulted from a true low prevalence of the agents, the failure to test for many agents or the low sensitivity of the diagnostic methods applied. Our meta-regression analysis of study and population variables showed that diagnostic methods determined the PMr estimates of typhoidal Salmonella and Dengue virus. An increase in the PMr of Klebsiella spp. infections was observed over time. Furthermore, the status of patients as either inpatient or outpatient predicted the PMr of Haemophilus spp. infections. CONCLUSION: The small number of epidemiological studies and the variety of NMFI agents on the African continent emphasizes the need for harmonized studies with larger sample sizes. In particular, diagnostic procedures for NMFIs should be standardized to facilitate comparability of study results and to improve future meta-analyses. Reliable NMFI burden estimates will inform regional public health strategies.

Seroprevalence and Associated Risk Factors of Trichinellosis and T. Solium Cysticercosis in Indigenous Pigs in Hoa Binh Province, Vietnam.

Trichinellosis and cysticercosis remain challenges to human health and animal productivity worldwide, especially in developing countries. While information on the occurrence of both diseases is infrequent, they are endemic in parts of Vietnam and mainly related to indigenous pigs kept by ethnic minorities. This study aimed to determine the seroprevalence and risk factors of both diseases in indigenous pigs and explore the perception and awareness of both human and pig trichinellosis and cysticercosis of pig farmers. A total of 352 pig sera samples from 131 holdings were collected and analyzed using ELISA antibody tests in six communes in the Da Bac districts of Hoa Binh province, Vietnam. A survey was conducted with representatives from these households to understand the knowledge and perspective on food-borne parasitic diseases. Overall, the seroprevalence of trichinellosis and T. solium cysticercosis was 13.6% (95% CI 10.2-17.7) and 1.7% (95% CI 0.6-3.7), respectively. The seroprevalence of trichinellosis was significantly higher in female and older pigs. Risk perception and knowledge of interviewed people on both human and pig trichinellosis and cysticercosis of pig farmers was poor. Risky practices, including free roaming of pigs and eating undercooked or fermented pork, were observed. Educational and awareness campaigns aligned with further research on feasible practice changes are critical to addressing these issues.