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Wireless capsule endoscopy is a non-invasive medical imaging modality used for diagnosing and monitoring digestive tract diseases. However, the analysis of images obtained from wireless capsule endoscopy is a challenging task, as the images are of low resolution and often contain a large number of artifacts. In recent years, deep learning has shown great promise in the analysis of medical images, including wireless capsule endoscopy images. This paper provides a review of the current trends and future directions in deep learning for wireless capsule endoscopy. We focus on the recent advances in transfer learning, attention mechanisms, multi-modal learning, automated lesion detection, interpretability and explainability, data augmentation, and edge computing. We also highlight the challenges and limitations of current deep learning methods and discuss the potential future directions for the field. Our review provides insights into the ongoing research and development efforts in the field of deep learning for wireless capsule endoscopy, and can serve as a reference for researchers, clinicians, and engineers working in this area inspection process., Competing Interests: No competing interests were disclosed., (Copyright: © 2024 Habe TT et al.)

Background: Cervical cancer is the leading cause of disability and mortality among women in Africa. Despite a significant correlation between HIV/AIDS and cervical cancer, there is unacceptably low coverage of the uptake of cervical cancer screening among human immunodeficiency virus-positive women in Sub-Saharan Africa. Individual primary studies are limited in explaining the patterns of uptake of cervical cancer screening. This review therefore considers the uptake of cervical cancer screening and its barriers among human immunodeficiency virus-positive women in Sub-Saharan Africa., Methods: We systematically searched articles published until December 31 , 2019, from the PubMed, Cochrane Library, POP LINE, Google Scholar, African Journals Online and JURN databases. The quality of the included articles was assessed by using the Newcastle‒Ottawa Scale, and the coverage of uptake of cervical cancer screening was pooled after checking for heterogeneity and publication bias. The random effect model was used, and subgroup analysis estimates were performed by country., Results: Twenty-one studies comprising 20,672 human immunodeficiency virus-positive women were included. Applying a random effect model, the overall cervical cancer screening uptake among this group of women in Sub-Saharan Africa was estimated to be 30% (95% CI: 19, 41, I 2  = 100%). The main barriers to uptake of cervical screening include poor knowledge about cervical cancer and screening, low risk perception of cervical cancer, fear of test result and fear of screening as painful, lack of access to screening services, high cost of screening service, and poor partner attitude and acceptance of the service. The perception of an additional burden of having a cervical cancer diagnosis was found to be a unique barrier among this population of women., Conclusion: The unacceptably low coverage of uptake of cervical cancer screening would indicate that the need to scale up the opportunities to these groups of women as well. This review revealed that in addition to structural and health care system barriers, sociocultural and personal barriers are powerful barriers in HIV-positive women. For these cohorts of population, a particular obstacle was discovered to be perception of an additional burden of having cervical cancer., (© 2023. The Author(s).)

Background: Implanon® is a commonly used effective long-acting reversible contraceptive method. It provides contraception for up to three years. Its early discontinuation was associated with an unwanted pregnancy, abortion, and socioeconomic burden. Thus, the main aim of this systematic review and meta-analysis is to determine the rate of early discontinuation of Implanon® and associated factors in Ethiopia., Method: This Systematic review and meta-analysis was performed by using online databases including PubMed, Google Scholar, Cochrane, HINARI, Web of Science, and other gray and online repositories of Ethiopian Universities. The JOANNA Briggs Institute standard data extraction and appraising sheet format was used for the extraction of all included studies. To test the heterogeneity of the studies the Cochran Q test and I 2 statistics test were used. The Funnel plot and Egger's tests were used to detect possible publication biases of the included studies. The forest plots were used to present the finding of the overall prevalence of the early Implanon® discontinuation and the odds ratio (OR) along with a 95% CI., Result: In this systematic review and meta-analysis seven studies with a total population of 3161 women using Implanon® were included. The overall pooled early Implanon® discontinuation rate was 31.34% (95%CI: 19.20, 43.47). Early discontinuation of Implanon® was associated with lack of counseling during service delivery 2.55times (OR: 2.55, 95%CI: 1.99, 3.25), the experienced side effect 3.25 times (OR: 3.25, 95%CI: 2.48, 4.24), absence of appointment after insertion 6.06 times (OR: 6.06, 95%CI: 2.15, 17.05), others decision on the women's choice 3.30 times (OR = 3.30, 95%CI: 2.52, 4.32), and lack the satisfaction of provided service 2.68 times (OR: 2.68, 96%CI: 1.61, 4.45)., Conclusion: About one-third of the women in Ethiopia discontinue the use of Implanon® within one year of the insertion. This is high compared to findings from other countries. Lack of counseling about the service, women's experience of the side effect, absence of the appointment following the service provision, other decisions on the method chosen, and lack of satisfaction were factors associated with the discontinuation of Implanon®. Hence, efforts should be made to reduce the rate of early discontinuation of Implanon® through drafting national guidelines and strategies accompanied by appropriate implementation, follow-up to foster adequate counseling, arrangement of appointments, helping women to decide on the choice, and increase the quality of care provision to enhance the satisfaction of the service., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper, (© 2023 Published by Elsevier Ltd.)

The need to make clean water accessible and affordable for low-income countries is crucial. This study examines the suitability of various clays for Ceramic Pot Water Filters production and groundwater treatment for effective microbe and fluoride removal. For this study, three clays were collected from different geographical locations in Ethiopia,i.e., Hosaenna Clay, Babawuha Clay, and Leku Clay. Organic additives such as sawdust and eragrostis tef husks were used to increase the porosity of the Ceramic Pot Water Filters. The Atterberg limit and particle size distribution tests revealed that BC and HC have moderate to high plasticity and mouldability, making them suitable for CPWF production. The clay chemical composition, phase analysis, and thermal properties were determined using XRF, XRD, and TGA/DTA. The turbidity, fluoride level, total dissolved solids, and pH of the groundwater decreases, from 13 to 0.45 NTU, from 3.4 to 0.053 mg/100 mL, from 1245 to 360 mg/l, and from 8.4 to 7.3, respectively; all of which are within the acceptable range of WHO drinking water standards. Microbial removal tests show that the CPWFs removed 99.3%-100% of total coliform bacteria and 98.48%-100% of fecal coliform bacteria from groundwater. Therefore, this work paves the way to fabricate a clay-based ceramic water filter for low-income countries to provide affordable household groundwater treatment technology for microbial and excess fluoride removal., Competing Interests: The authors declare no conflict of interest., (© 2023 The Authors.)

Elastin and collagen were isolated from waste broil skin and modified with l-lactic acid oligomer for the efficacy of substituting petroleum based ointment base matrix. Paraffin wax, which is one of the most extensively used petroleum ointment derivative is well known for its incompatibility with the skin. Chronically it clogs the skin pores, which subsequently affects the release of moisture. To mitigate this problem, a novel approach has been followed to synthesize a fully green and biocompatible ointment base matrix. The extracted Elastin and Collagen (which exists naturally in our skin) and lactic acid monomer (commonly known as a part of a body fluid) mixed at different proportion and used to synthesize a lactic acid modified elastin/collagen (OLLA-m-ELN/COL) bioconjugate matrix with lubricating characteristics via in-situ polycondensation reaction. The macromolecular interaction between Amide I group of elastin/collagen and C=O of a lactic acid oligomer was confirmed by FTIR analysis. Organoleptic analysis, spreadability, pH, and viscosity were analyzed. The six-mass loss stages which was observed in the ELN/COL matrix was changed into a single mass loss for the synthesized bioconjugate with improved thermal stability. The thermal stability improvement can be correlated with the formation of secondary macromolecular interaction., Competing Interests: The authors declare no conflict of interest., (© 2022 The Author(s).)

COVID-19 pandemic caused havoc around the globe in both economic and non-economic sectors. This paper, unlike previous studies, evaluates the role of COVID-19 on the volatility in natural resources. The volatility of natural resources commodity prices has been the center of discussion, especially during the pandemic. Unlike previous studies, this study aims to evaluate the role of the pandemic, i.e., Covid-19 and its possible impact on volatility in natural resources commodity prices for China. China has been the center of this epidemic disease and is considered one of the major economies affected by the Covid-19; therefore, it is better to conduct this study for China. This study uses data from January 2020 till September 2021 to capture the peak time of Covid-19. Moreover, this study employs the novel wavelet power spectrum and wavelet coherence approach to better capture volatility within commodity prices volatility and Covid-19 and evaluate the association between both variables. The empirical results reveal that only natural resources commodity prices are volatile and only short. While Covid-19 positive cases and Covid-19 deaths are not vulnerable during the study period. Moreover, the wavelet coherence conforms that both Covid-19 positive cases and Covid-19 deaths significantly cause volatility in natural resources commodity prices. Although, volatility is found at different periods; still, volatility is observed only in the short-run. The study also provides relevant policy implications to ensure a relevant and timely solution for the existing issue. Moreover, future research guidelines and the study's limitations are also provided., (© 2022 Published by Elsevier Ltd.)

Ready-to-eat (RTE) salads and berries are increasingly consumed in industrialized countries. These products can be contaminated by pathogenic parasites that have been responsible for foodborne outbreaks worldwide. In Italy, there are few data on contamination of RTE salads and berries with parasite transmission stages and this requires more-in-depth investigations. To estimate the prevalence of contamination with Cryptosporidium spp. and Giardia duodenalis in these fresh products, a total of 324 packages of local RTE mixed salads - belonging to three different industrial brands - and 324 packages of berries - blueberries from Peru, blackberries from Mexico, raspberries from Italy - were bought from supermarkets located in the Provinces of Bari and Foggia, Apulia, Italy. A pool size of nine packages was chosen and a total of 72 pools were processed in the whole year. After washing, the pellets were examined by microscopy (FLOTAC) and tested using conventional simplex PCR, targeting Cryptosporidium spp., Giardia duodenalis, and Entamoeba spp., and sequencing. Several Cryptosporidium species and Giardia duodenalis assemblages, some of which are of potential zoonotic relevance, as well as Entamoeba spp., were identified in both matrices. By microscopy, Giardia-like cysts in local raspberries and Entamoeba-like cysts in imported blueberries were detected. Giardia duodenalis (Assemblages A, B and E) and Entamoeba histolytica were molecularly confirmed with overall prevalences of 4.6% (95% C.I. 3.0-6.8) and 1% (95% C.I. 0.3-2.1), respectively. Molecular methods identified Cryptosporidium ryanae, Cryptosporidium bovis, Cryptosporidium xiaoi, and Cryptosporidium ubiquitum in both matrices, with a prevalence of 5.1% (95% C.I. 3.3-7.3). A distinct seasonality in prevalence was observed for G. duodenalis, with most positives occurring in spring, whereas Cryptosporidium showed no significant seasonal variations. These results highlight that inadequate management of fresh produce, both locally produced and imported, along the food chain may have the potential for consequences on human health., (Copyright © 2022 Elsevier B.V. All rights reserved.)

The risk of foodborne parasite infection linked to the consumption of contaminated fresh produce has long been known. However, despite epidemiological links between the outbreaks and contaminated berries, few studies have assessed the magnitude of parasite contamination on fresh produce sold in Europe. The present study was aimed to address the knowledge gap on parasite contamination of berries sold in Norway. Samples of blueberries, strawberries, and raspberries were analysed by multiplex qPCR for detection of Echinococcus multilocularis, Toxoplasma gondii, and Cyclospora cayetanensis. In addition, a simplex qPCR method was employed for detecting contamination of the berries with Cryptosporidium spp. A total of 820 samples of berries, each of around 30 g (274 samples of blueberries, 276 samples of raspberries, and 270 samples of strawberries), were analysed. We found an overall occurrence of 2.9%, 6.6%, and 8.3% for T. gondii, C. cayetanensis, and Cryptosporidium spp., respectively, whereas E. multilocularis was not detected from any of the samples investigated. Strawberries and raspberries were most often contaminated with Cryptosporidium spp., whereas blueberries were contaminated mostly with C. cayetanensis. Detection of parasite contaminants on fresh berries indicates the need for a system to ensure the parasitological safety of fresh berries., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

COVID-19 affected the economies of both developed and developing countries through multiple channels. However, the impact of this pandemic was also not so different for the developed world too. This study aims to evaluate the effect of the natural resources' rents such as oil, natural gas and energy rents on the economic performance of G7 economies from 1990 to 2020. This study uses updated panel data methods to identify the effect of COVID-19 by using novel diagnostic and unit root methods. The study found that during the COVID-19 and post pandemic, economic performance has been declined. This study found that natural resources rent, including oil and gas, help to improve the economic performance. Further, structural break for the year 2019 caused by COVID-19 also slowed down the economic performance of G7 economies. This study suggests more stabilization policies towards natural resources commodity prices and encouragement of active role from the G7 countries. This study further provides relevant policy implications in the concluding section for the selected group of countries., (© 2021 Published by Elsevier Ltd.)

Cracks in typical mortar constructions enhance water permeability and degrade ions into the structure, resulting in decreased mortar durability and strength. In this study, mortar samples are created that self-healed their cracks by precipitating calcium carbonate into them. Bacillus subtilus bacterium (10 -7 , 10 -9 cells/mL), calcium lactate, fine aggregate, OPC-cement, water, and bagasse ash were used to make self-healing mortar samples. Calcium lactates were prepared from discarded eggshells and lactic acid to reduce the cost of self-healing mortars, and 5% control burnt bagasse ash was also employed as an OPC-cement alternative. In the presence of moisture, the bacterial spores in mortars become active and begin to feed the nutrient (calcium lactate). The calcium carbonate precipitates and plugs the fracture. Our experimental results demonstrated that cracks in self-healing mortars containing bagasse ash were largely healed after 3 days of curing, but this did not occur in conventional mortar samples. Cracks up to 0.6 mm in self-healing mortars were filled with calcite using 10 -7 and 10 -9 cell/mL bacteria concentrations. Images from an optical microscope, X-ray Diffraction (XRD), and a scanning electron microscope (SEM) were used to confirm the production of calcite in fractures. Furthermore, throughout the pre- and post-crack-development stages, self-healing mortars have higher compressive strength than conventional mortars. The precipitated calcium carbonates were primed to compact the samples by filling the void spaces in hardened mortar samples. When fissures developed in hardened mortars, bacteria became active in the presence of moisture, causing calcite to precipitate and fill the cracks. The compressive strength and flexural strength of self-healing mortar samples are higher than conventional mortars before cracks develop in the samples. After the healing process of the broken mortar parts (due to cracking), self-healing mortars containing 5% bagasse ash withstand a certain load and have greater flexural strength (100 kPa) than conventional mortars (zero kPa) at 28 days of cure. Self-healing mortars absorb less water than typical mortar samples. Mortar samples containing 10 -7 bacteria cells/mL exhibit greater compressive strength, flexural strength, and self-healing ability. XRD and SEM were used to analyze mortar samples with healed fractures. XRD, FTIR, and SEM images were also used to validate the produced calcium lactate. Furthermore, the durability of mortars was evaluated using DTA-TGA analysis and water absorption tests.

Porcelain insulators prepared from locally available ceramic materials, such as Hosaena clay, quartz, partially replaced feldspar by cullet (rich in alkaline materials). The physical and electrical properties, such as water absorption, apparent porosity, bulk density, linear shrinkage, and dielectric strength as a function of firing temperature, were evaluated. The XRD and AAS measurement results show Hosaenna clay mainly belongs to kaolinite minerals with a sufficient SiO2 (45.60wt. %), Al2O3 (35.52wt. %), and it possesses reasonable plasticity nature. The SEM and Electrical properties measurement shows the porcelain insulators fired at 1150 °C & 1200 °C showed homogenized primary and secondary mullite with optimum glassy phase and good dielectric strength for practical applications. The primary & secondary mullite and glassy phases were grown early at a firing temperature of 1150 and 1200 °C, due to the partial replacement of feldspar by cullet. The porcelain insulator prepared from the composition of 45%clay, 35%feldspar, 10%quartz and 10%cullet and fired at 1200 °C showed dielectric strength 8.9kv/mm, water absorption (0.459%), apparent porosity (1.530%), bulk density (4.29 g/cm 3 ), and linear shrinkage (4.18%). Therefore, incorporating economic alkaline materials such as cullet to improve the alkalinity of the locally available clay materials, Hosaenna clay, Arero feldspar, and Arero quartz reduces the firing temperature with excellent dielectric strength., Competing Interests: The authors declare no conflict of interest., (© 2022 The Authors. Published by Elsevier Ltd.)

An episiotomy is one of the most commonly performed obstetrics surgeries indicated in emergencies during the second stage of labor like fetal distress, dystocia, and tight perineum. As a result, this systematic review and meta-analysis aimed to assess the prevalence of episiotomy practice and associated factors in Ethiopia. Ten cross-sectional studies with a total population of 3718 were included in this study. The search was done using online databases like PubMed, HINARI, Web of Science, other gray, and online repositories of Universities. All the included papers were extracted and appraised using the standard extraction sheet format of JOANNA Briggs Institute. The Cochran Q-test and I 2 statistics test were used to test the heterogeneity of studies. To detect the publication bias of the included studies, a funnel plot and Egger's test were used. The pooled prevalence of episiotomy practice and the odds ratio with a 95% confidence interval were presented using forest plots. The overall pooled prevalence of episiotomy practice was 45.11% (95% CI; 37.04-53.18; I 2  = 96.3%). Prolonged second stage of labor (OR: 4.79, 95% CI: 3.03, 7.57), face presentation (OR: 4.26, 95% CI: 1.21, 15.07), birth weight > 4000 g (OR: 6.71, 95% CI: 3.14-14.33), instrumental delivery (OR: 4.26, 95% CI 2.95, 6.14), and primiparity (OR: 3.70, 95% CI: 1.90, 7.2) were factors associated with episiotomy practice. The overall prevalence of episiotomy practice was higher in Ethiopia compared to studies conducted in other countries. The prolonged second stage of labor, face presentation, birth weight > 4000 g, instrumental delivery, and primiparity of women were the factors associated with episiotomy practice. Therefore, efforts should be made to prevent routine episiotomy practice through creating awareness, adjusting national guidelines, affecting the World Health Organization episiotomy policy, and monitoring the activities of the health care facilities in executing the protocols.

To investigate the presence of Cyclospora cayetanensis, Toxoplasma gondii and Echinococcus spp. in fresh produce sold in Italy, 324 locally produced 'ready-to-eat' (RTE) mixed-salad packages belonging to three brands and 324 berries packages (blueberries and blackberries imported from Peru and Mexico, respectively, and raspberries grown in Italy) were purchased at retail. Nine individual packages from each of the six types of fresh produce were collected monthly for one year, and with the same produce pooled, this resulted in a total of 72 pools for the whole year. Using microscopy (FLOTAC), a Cyclospora-like oocyst was detected in a blueberry sample and a taeniid egg was detected in a RTE-salad sample. Molecular tools confirmed these to be C. cayetanensis and Echinococcus multilocularis, respectively. Toxoplasma gondii was not detected in any of the samples. This study shows for the first time in Europe that imported berries on the Italian market may be contaminated with C. cayetanensis and RTE salads grown in Italy with E. multilocularis. The results indicate a new epidemiological scenario and highlight that current management of fresh produce, locally produced or imported, does not ensure products are free from parasite contamination., (Copyright © 2021. Published by Elsevier Ltd.)

Cryptosporidium oocysts are known for being very robust, and their prolonged survival in the environment has resulted in outbreaks of cryptosporidiosis associated with the consumption of contaminated water or food. Although inactivation methods used for drinking water treatment, such as UV irradiation, can inactivate Cryptosporidium oocysts, they are not necessarily suitable for use with other environmental matrices, such as food. In order to identify alternative ways to inactivate Cryptosporidium oocysts, improved methods for viability assessment are needed. Here we describe a proof of concept for a novel approach for determining how effective inactivation treatments are at killing pathogens, such as the parasite Cryptosporidium. RNA sequencing was used to identify potential up-regulated target genes induced by oxidative stress, and a reverse transcription quantitative PCR (RT-qPCR) protocol was developed to assess their up-regulation following exposure to different induction treatments. Accordingly, RT-qPCR protocols targeting thioredoxin and Cryptosporidium oocyst wall protein 7 (COWP7) genes were evaluated on mixtures of viable and inactivated oocysts, and on oocysts subjected to various potential inactivation treatments such as freezing and chlorination. The results from the present proof-of-concept experiments indicate that this could be a useful tool in efforts towards assessing potential technologies for inactivating Cryptosporidium in different environmental matrices. Furthermore, this approach could also be used for similar investigations with other pathogens.

Due to the delicate nature of berries and the reduced shelf-life once washed, producers usually do not wash berries. Therefore, consumers are expected to wash the berries prior to consumption, and this might be a more effective way of infection prevention. However, the efficacy of consumer berry-washing procedures in removing the parasite contaminants from the berries surface has not been investigated. The aim of the present study was, therefore, to compare the efficacy of three different washing techniques in removing parasite contaminants. Three alternatives to washing berries before consumption were compared on berries artificially contaminated with oo/cysts of Cyclospora cayetanensis , Cryptosporidium parvum , and Giardia duodenalis . The results show that simple washing of berries under the cold tap for 1 min could remove on average at least 80% of the parasites, except for C. cayetanensis , which seems to be stickier than both G. duodenalis and C. parvum . The percent removal was slightly lower for raspberries as compared to blueberries. Although the differences are expected, a relevant result of the study is that washing contaminated berries prior to consumption by the consumer removes a considerable proportion of parasites and thereby lowers the risk of ingesting parasites' transmission stages.

Abstract: Berries are potential vehicles for the transmission of parasites and have been implicated in illness outbreaks in various countries around the world, particularly in the United States. Although data on contamination of fresh produce with foodborne parasites have been obtained from various global regions, data from Colombia are lacking even though South American countries are major producers of fresh produce, which is both consumed nationally and exported. In this study, we used a previously published multiplex quantitative PCR approach to investigate contamination of strawberries purchased in either supermarkets or local markets in 20 localities. Strawberries were washed in a detergent solution after purchase, and the eluate was concentrated and sent to Norway for molecular analysis. Of the 120 strawberry samples analyzed, wash eluate from 6 samples (5%) tested positive for Toxoplasma gondii DNA, and 1 sample (0.83%) was positive for Cyclospora cayetanensis DNA. These results indicate that strawberries for sale in Bogotá, Colombia, may be contaminated with T. gondii and C. cayetanensis and, therefore, could act as transmission vehicles for these parasites. These data also indicate that cat and human fecal contamination of the strawberries has occurred at some point in their production, transportation, or storage. These findings highlight the need for a hazard analysis critical control point investigation of the strawberry production chain and implementation of measures to reduce the risk of strawberry contamination, thereby minimizing the risk of transmission of parasitic infection via these fruits, which are usually consumed raw., (Copyright ©, International Association for Food Protection.)

The potential public health impact of foodborne parasites (FBP) transmitted via contaminated fresh produces indicates the necessity for robust and reliable laboratory methods for their detection and identification on this infection vehicle. Standardization of methods for detection of common FBP in fresh produce is to be expected and ensuring that the DNA extraction approach is most appropriate for the FBP of interest and for the matrix being analyzed is also important. Therefore, the aim of the present study was to compare the efficacy of two commercially available DNA extraction procedures, the UNEX-based method and DNeasy PowerSoil kit in the detection of three protozoan parasites, C. cayetanensis, C. parvum, and T. gondii, on contaminated berries. Oocysts of each parasite were spiked into the pellets of raspberry and blueberry washes. The spiked pellets were then randomly assigned to DNA extraction using either the PowerSoil or UNEX method, with DNA extraction with both methods performed by two independent analysts. The detection rate when berry washes were spiked with 20 oocysts of C. cayetanensis, T. gondii, and C. parvum was 95%, 85%, and 40%, respectively, when using the PowerSoil kit; whereas the equivalent results using the UNEX method were 55%, 60%, and 5%, respectively. In addition, significantly lower C q values were achieved for each parasite in the samples spiked with 500 oocysts when the PowerSoil kit was used. Possible reasons for these results are discussed, and include the composition of both the beads and the buffers in each method., (Copyright © 2020 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Foodborne parasites (FBP) are of major public health importance and warrant appropriate detection and control strategies. Most of the FBP considered for risk-ranking by a panel of experts are potentially transmitted via consumption of contaminated fresh produce, including berries. In this study we focused on the potential of three FBP, namely Echinococcus multilocularis, Toxoplamsa gondii, and Cyclospora cayetanensis, as contaminants of berries. Surveys to assess these parasites as contaminants of fresh produce in general, and berries in particular, are scanty or non-existent mainly due to the lack of optimized laboratory methods for detection. The aim of the present study was to develop and evaluate a novel multiplex qPCR for the simultaneous detection of E. multilocularis, T. gondii, and C. cayetanensis from berry fruits. The efficiency and linearity of each channel in the multiplex qPCR were within the acceptable limits for the range of concentrations tested. Furthermore, the method was shown to have good repeatability (standard deviation ≤0.2 C q ) and intermediate precision (pooled standard deviation of 0.3-0.6 C q ). The limit of detection was estimated to 10 oocysts for Toxoplasma and Cyclospora, and 5 eggs for Echinococcus per 30 g of raspberries or blueberries. In conclusion, evaluation of the present method showed that the newly developed multiplex qPCR is highly specific, precise, and robust method that has potential for application in food-testing laboratories., (Copyright © 2019 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Cyclospora cayetanensis is a coccidian parasite that is associated with foodborne outbreaks of gastrointestinal illnesses. Raspberries have been implicated as a vehicle of infection in some of these outbreaks. Most of the molecular techniques used for the detection of parasites commonly use the 18s rRNA as a target gene, which is highly conserved. The conserved nature of the 18s rRNA gene among coccidia means that there is potential for cross-reactivity from primers intended to target this gene in C. cayetanensis with the same gene in related coccidia. This provides an additional challenge in developing a specific detection method. The aim of this study is to develop a new, more specific assay to detect C. cayetanensis in berry fruits. This new assay, targeting the internal transcribed spacer 1 (ITS-1) region, was tested on three different berry matrices: raspberries, blueberries, and strawberries. The new assay showed good efficiency (102%), linearity ( r 2 = 0.999), repeatability (standard deviation of C q 0.2 (95% CI: 0.2, 0.3) and specificity for Cyclospora , with no cross-reactivity with related coccidia ( Toxoplasma gondii, Eimeria mitis, Cystoisospora canis , and Cryptosporidium parvum ) when tested in vitro . The method development was initially conducted using Cyclospora DNA only. After it was confirmed to have an acceptable performance, the method was evaluated using the oocysts of C. cayetanensis . The method was also improved by incorporating an internal control as a duplex in order to monitor PCR inhibition due to sample matrix components. The duplex assay also showed a good efficiency (100%) and linearity ( r 2 = 0.99). The results showed that the new assay has potential for standard use in food testing laboratories. Furthermore, results regarding important factors related to assay robustness are discussed.

In the autumn of 2018, an outbreak of cryptosporidiosis affected adult employees from the same company in Western Norway. The organism was Cryptosporidium parvum, GP60 subtype IIaA14G1R1. All those infected had drunk from the same container of self-pressed apple juice. Incubation period (1 week) and clinical signs were similar among those infected, although some experienced a more prolonged duration of symptoms (up to 2-3 weeks) than others. The infections resulted after consumption from only one of 40 containers of juice and not from any of the other containers. It seems that although Cryptosporidium oocysts were detected in a sample from another container, the contamination did not affect the whole batch. This is perhaps indicative of a restricted contamination event, either from contaminated ground in the orchard, or during collection of the fruit, or during processing. Although outbreaks of food-borne cryptosporidiosis have previously been associated with consumption of contaminated apple juice, most of the more recent outbreaks of food-borne cryptosporidiosis have been associated with salad vegetables or herbs. This outbreak, the first outside USA reported to be associated with apple juice, is a timely reminder that such juice is a suitable transmission vehicle for Cryptosporidium oocysts, and that appropriate hygienic measures are essential in the production of such juice, including artisanal (non-commercial) production.