Neurocysticercosis Diagnosis in a Non-Endemic Country: France.

Diagnosing neurocysticercosis (NCC) is difficult due to its variable clinical presentations and the different imaging techniques used to detect brain damage. This study aimed to evaluate the use of cerebrospinal fluid serology and PCR for diagnosing biological neurocysticercosis in a non-endemic country. We tested samples from patients living in France with suspected NCC and confirmed that 45 of the patients presented with the disease. A total of 89% of patients had previously traveled to countries where the disease was endemic. The sensitivity of Western blots compared to ELISA was not significantly different (80% vs. 60%) (p > 0.05), and neither was the sensitivity of Western blots vs. PCR (78% vs. 56%) (p > 0.05). The PCR sensitivity was 78% and 47% in definitive NCC and in probable NCC. PCR tests using cerebrospinal fluid should be considered as a diagnostic criterion for identifying NCC.

Unveiling the incidences and trends of the neglected zoonosis cystic echinococcosis in Europe: a systematic review from the MEmE project.

The neglected zoonosis cystic echinococcosis affects mainly pastoral and rural communities in both low-income and upper-middle-income countries. In Europe, it should be regarded as an orphan and rare disease. Although human cystic echinococcosis is a notifiable parasitic infectious disease in most European countries, in practice it is largely under-reported by national health systems. To fill this gap, we extracted data on the number, incidence, and trend of human cases in Europe through a systematic review approach, using both the scientific and grey literature and accounting for the period of publication from 1997 to 2021. The highest number of possible human cases at the national level was calculated from various data sources to generate a descriptive model of human cystic echinococcosis in Europe. We identified 64 745 human cystic echinococcosis cases from 40 European countries. The mean annual incidence from 1997 to 2020 throughout Europe was 0·64 cases per 100 000 people and in EU member states was 0·50 cases per 100 000 people. Based on incidence rates and trends detected in this study, the current epicentre of cystic echinococcosis in Europe is in the southeastern European countries, whereas historical endemic European Mediterranean countries have recorded a decrease in the number of cases over the time.

Severe and Rare Case of Human Dirofilaria repens Infection with Pleural and Subcutaneous Manifestations, Slovenia.

We report a case of human Dirofilaria repens infection in a woman in Slovenia who had concomitant pleural and subcutaneous manifestations of the infection. This case report illustrates the clinical course of a severe symptomatic parasitic infection that had multisystemic manifestations.

Alveolar echinococcosis in nutria (Myocastor coypus), invasive species in Slovenia.

The present report describes a case of Echinococcus multilocularis infection in nutria (Myocastor coypus) culled in the central area of Slovenia. Post-mortem exam showed multiple cystic lesions in the liver. Gross examination, as well as parasitological and histopathological examinations, revealed numerous cysts of various sizes, filled with yellow clear fluid and displacing most of the liver parenchyma. The cyst lumina contained numerous protoscolices approximately 100 µm in diameter and calcareous corpuscles. The protoscolices had two visible suckers and a rostellum with birefringent hooks. The lesions were consistent with an E. multilocularis cyst. Molecular analysis confirmed that the nutria was infected with E. multilocularis. To our knowledge, this is the first report of echinococcosis in nutria in Slovenia that presents gross, parasitological, and histological lesions and the result of molecular analysis. Nutrias in Slovenia are dangerous invaders but can also be a relevant bioindicator of the presence of the parasite in the environment.

Diagnostic challenge of Strongyloides stercoralis hyperinfection syndrome: a case report.

Strongyloides stercoralis causes chronic, mostly asymptomatic infections but hyperinfection syndrome may occur in immunosuppressed patients, especially in those receiving corticosteroids. We report a case of S. stercoralis hyperinfection syndrome in a solid organ transplant recipient that occurred approximately 2.5 months after heart transplantation. The patient presented to the intensive care unit with acute respiratory distress, bacteremia, and petechial rash on abdomen and toe. Microbiology testing of respiratory samples excluded infection with Pneumocystis jirovecii, respiratory viruses, pathogenic bacteria and fungi. No eosinophilia was found. Histopathological examination of the skin biopsy of the petechial rash provided the first indication of the diagnosis, revealing the presence of isolated filariform S. stercoralis larvae in the dermis. Subsequent microbiology testing confirmed the diagnosis. This case highlights the role of histopathological examination of a skin rash in diagnosing patients with atypical clinical presentation of Strongyloides hyperinfection syndrome.

Dirofilaria Repens in Dogs and Humans in Slovenia.

Introduction: The prevalence of Dirofilaria repens in dogs in countries bordering Slovenia ranges from 1.5% to 47.3%. The aim of this study was to estimate its prevalence in Slovenian dogs and to present the cases of dirofilariasis diagnosed in humans from 2010 to 2020. Material and Methods: Epidemiological data were collected and blood samples were taken from 465 dogs older than one year and born in Slovenia. A real-time PCR was performed on all samples to detect filarioid DNA, and a D. repens-and D. immitis-specific real-time PCR was performed on positive samples. Blood samples from 446 dogs were tested for Dirofilaria spp. using a modified Knott's test. Human cases were diagnosed from histological sections of excised subcutaneous nodules. Descriptive statistics were used to characterise the samples. The one-sample nonparametric chi-squared test was used to assess whether categories of a variable were equally distributed. Results: Three dogs' samples tested positive for D. repens using the species-specific real-time PCR, while D. immitis DNA was not detected. The modified Knott's test was positive in two of the three PCR-positive dogs, two of which had never travelled outside Slovenia's borders. Four human patients with D. repens dirofilariasis were diagnosed. Since their travel history was unknown, autochthonous transmission could not be confirmed. Conclusion: Our study demonstrated a 0.64% prevalence of D. repens infection in dogs in Slovenia. Two cases could be autochthonous.

Canine Leishmaniasis Prevalence in the Slovenian Dog Population.

INTRODUCTION: Leishmaniasis is a life-threatening zoonosis of which dogs are the major reservoir and sandflies are the vectors. Until now, the prevalence of canine leishmaniasis (CanL) in the Slovenian dog population was unknown. MATERIAL AND METHODS: Epidemiological data, eye swabs and blood samples were taken from 465 dogs born in Slovenia and older than one year. Commercial ELISA kits and real-time PCR were used. For ELISA-positive samples, an immunofluorescence antibody test (IFAT) was performed. Descriptive statistics were used to characterise the samples. The one-sample nonparametric chi-square test was used to test whether the categories of a variable were equally distributed. RESULTS: A 59.9% proportion of the recruited dogs had travelled to endemic regions and 62.1% of them had not been protected by insect repellents. Skin symptoms that might be CanL-related were described in 109 of the dogs' histories (23.4%), inappetence and/or weight loss in 25 (5.4%), and anaemia, intermittent fever, and/or lymphadenopathy in 19 (4.1%). At the time of recruitment, all dogs were asymptomatic. All samples were PCR negative, nine (1.9%) were ELISA positive, but none were IFAT positive. Five of the nine ELISA-positive dogs were non-travellers. CONCLUSION: We conclude that the seroprevalence of canine leishmaniasis of 1.9 % in the autochthonous Slovenian dog population may pose a risk of endemic spread of the disease.

Sarcocystis species in bovine carcasses from a Belgian abattoir: a cross-sectional study.

BACKGROUND: Sarcocystis species are obligatorily heteroxenous parasites, of which some are zoonotic, representing a public health and economic impact. This study investigated the occurrence of Sarcocystis spp. in cattle sampled from a Belgian slaughterhouse. METHODS: A total of 200 carcasses were included in the study, sampled during 10 sampling days. The sedimentation method was applied to isolate the sarcocysts from both heart and diaphragm muscles collected from each carcass. Multiplex PCR, PCR-RFLP as well as cox1 gene sequencing techniques were applied serially on collected sarcocysts for species identification. RESULTS: Sarcocystis spp. were detected in 64% (128/200; 95% CI 57-71%) of the sampled carcasses. Female dairy cattle presented the highest Sarcocystis occurrence rate (91%) as well as the highest Sarcocystis species diversity compared to female beef and male beef. Sarcocystis spp. were detected more often in the heart muscles than in the diaphragm among female beef (p < 0.001) and dairy carcasses (p = 0.001), while in male carcasses no significant difference was observed (p = 0.763). The effect of age was not significant in male carcasses (p = 0.872), while the odds of finding sarcocysts significantly increased with age (p = 0.003) within both types of female carcasses. S. cruzi was the most prevalent species and was found in 56.5% (113/200) of the carcasses, followed by S. hominis (21.0%, 42/200), S. bovifelis (12.5%, 25/200), S. bovini (2.0%, 4/200), S. hirsuta (1.5%, 3/200) and S. heydorni (0.5%, 1/200). Six different species were detected in the diaphragm, while only two species were recovered from the heart. S. cruzi was the most prevalent species in heart, while in the diaphragm, this was S. hominis. CONCLUSIONS: The detection of S. hominis in 21% of the sampled carcasses presents a potential food safety issue, and further research is warranted into controlling this infection.

Ignatzschineria larvae Bacteremia Following Lucilia sp. Myiasis in an Irregular Migrant: A Case Report.

Bacteremia induced by wound myiasis is uncommon and therefore rarely suspected by clinicians when treating patients with neglected wounds. We present a case of Ignatzschineria larvae bacteremia as a complication of Lucilia sp. maggot wound myiasis in a young male migrant. This is the first reported human case of Ignatzschineria bacteremia in Slovenia and one of the 2 described in the literature where the fly larvae infesting the wounds of the patient with Ignatzschineria bacteremia were not only suspected to be Lucilia sp. but also entomologically identified.

Molecular Characterization of Echinococcus granulosus sensu lato from Humans in Slovenia.

The larval form of tapeworms of the Echinococcus granulosus sensu lato species cluster cause an important zoonotic infection, cystic echinococcosis (CE). Molecular characterization of the cluster's isolates from different hosts greatly contributes to a better understanding of its transmission dynamics. To date, no genetic information is available on CE in Slovenia. In this work, we characterized isolates from human CE cases. Parasite samples from 18 patients were collected, together with the patients' demographic and clinical data. Genomic DNA was analyzed by conventional PCR and sequencing at four mitochondrial loci (cytochrome c oxidase subunit 1, cox1; NADH dehydrogenase subunit 1, nad1; NADH dehydrogenase subunit 5, nad5; and small ribosomal RNA, rrnS). Thirteen isolates were successfully amplified and sequenced. Seven (58.8%) patients were infected with E. granulosus sensu stricto (s.s.) G1, five (38.5%) with E. canadensis G7 and one (7.7%) with E. granulosus s.s. G3. Echinococcus canadensis G7, the pig genotype, was identified exclusively in autochthonous Slovenes, while the patients originating from the Western Balkans were all infected with E. granulosus s.s. Our findings suggest that pigs are important intermediate hosts for human CE in Slovenia.

Human taeniasis/cysticercosis: a potentially emerging parasitic disease in Europe.

Taenia saginata (T. saginata)/Taenia solium (T. solium) taeniasis/cysticercosis disease complexes remain a significant challenge for food safety and public health. Human taeniasis is an infectious disease caused by the ingestion of the metacestode larval stage, the cysticerci of T. saginata in beef or T. solium in pork. Humans can also become infected via the ingestion of T. solium eggs. In this case, the cysticerci can establish in the central nervous system, causing the infection called neurocysticercosis. T. solium is of higher importance than T. saginata because the former species can cause neurocysticercosis in humans, a major cause of neurological morbidity in the world. The taeniasis/cysticercosis complex is included in the list of neglected zoonotic diseases by the World Health Organization and Food and Agriculture Organization, with T. solium being the number one foodborne parasite; it occurs mostly in developing countries, such as regions of Asia, Africa and Latin America, where the disease remains endemic. Long absent in Western Europe and other developed countries, cysticercosis has been recently re-emerged as a result of immigration, travel and commerce. In this review, cysticercosis is presented with special emphasis on some aspects of this neglected disease: the main clinical manifestations, risk factors and epidemiology. In addition, any recent advances in diagnostic approaches and treatment are discussed. Finally, the complexities involved in the control of the disease and the need to revise current management strategies are highlighted.

Review of Cryptosporidium and Giardia in the eastern part of Europe, 2016.

IntroductionThis paper reviews the current knowledge and understanding of Cryptosporidium spp. and Giardia spp. in humans, animals and the environment in 10 countries in the eastern part of Europe: Bosnia and Herzegovina, Croatia, Czech Republic, Estonia, Hungary, Latvia, Poland, Romania, Serbia and Slovenia. Methods: Published scientific papers and conference proceedings from the international and local literature, official national health service reports, national databases and doctoral theses in local languages were reviewed to provide an extensive overview on the epidemiology, diagnostics and research on these pathogens, as well as analyse knowledge gaps and areas for further research. Results:Cryptosporidium spp. and Giardia spp. were found to be common in eastern Europe, but the results from different countries are difficult to compare because of variations in reporting practices and detection methodologies used. Conclusion: Upgrading and making the diagnosis/detection procedures more uniform is recommended throughout the region. Public health authorities should actively work towards increasing reporting and standardising reporting practices as these prerequisites for the reported data to be valid and therefore necessary for appropriate control plans.

A European network for food-borne parasites (Euro-FBP): meeting report on 'Analytical methods for food-borne parasites in human and veterinary diagnostics and in food matrices'.

Food-borne parasites (FBPs) are a neglected topic in food safety, partly due to a lack of awareness of their importance for public health, especially as symptoms tend not to develop immediately after exposure. In addition, methodological difficulties with both diagnosis in infected patients and detection in food matrices result in under-detection and therefore the potential for underestimation of their burden on our societies. This, in consequence, leads to lower prioritization for basic research, e.g. for development new and more advanced detection methods for different food matrices and diagnostic samples, and thus a vicious circle of neglect and lack of progress is propagated. The COST Action FA1408, A European Network for Foodborne Parasites (Euro-FBP) aims to combat the impact of FBP on public health by facilitating the multidisciplinary cooperation and partnership between groups of researchers and between researchers and stakeholders. The COST Action TD1302, the European Network for cysticercosis/taeniosis, CYSTINET, has a specific focus on Taenia solium and T. saginata, two neglected FBPs, and aims to advance knowledge and understanding of these zoonotic disease complexes via collaborations in a multidisciplinary scientific network. This report summarizes the results of a meeting within the Euro-FBP consortium entitled 'Analytical methods for food-borne parasites in human and veterinary diagnostics and in food matrices' and of the joined Euro-FBP and CYSTINET meeting.

Epidemiology of taeniosis/cysticercosis in Europe, a systematic review: Western Europe.

BACKGROUND: Taenia solium and Taenia saginata are zoonotic parasites of public health importance. Data on their occurrence in humans and animals in western Europe are incomplete and fragmented. In this study, we aimed to update the current knowledge on the epidemiology of these parasites in this region. METHODS: We conducted a systematic review of scientific and grey literature published from 1990 to 2015 on the epidemiology of T. saginata and T. solium in humans and animals. Additionally, data about disease occurrence were actively sought by contacting local experts in the different countries. RESULTS: Taeniosis cases were found in twelve out of eighteen countries in western Europe. No cases were identified in Iceland, Ireland, Luxembourg, Norway, Sweden and Switzerland. For Denmark, Netherlands, Portugal, Slovenia, Spain and the UK, annual taeniosis cases were reported and the number of detected cases per year ranged between 1 and 114. Detected prevalences ranged from 0.05 to 0.27%, whereas estimated prevalences ranged from 0.02 to 0.67%. Most taeniosis cases were reported as Taenia spp. or T. saginata, although T. solium was reported in Denmark, France, Italy, Spain, Slovenia, Portugal and the UK. Human cysticercosis cases were reported in all western European countries except for Iceland, with the highest number originating from Portugal and Spain. Most human cysticercosis cases were suspected to have acquired the infection outside western Europe. Cases of T. solium in pigs were found in Austria and Portugal, but only the two cases from Portugal were confirmed with molecular methods. Germany, Spain and Slovenia reported porcine cysticercosis, but made no Taenia species distinction. Bovine cysticercosis was detected in all countries except for Iceland, with a prevalence based on meat inspection of 0.0002-7.82%. CONCLUSIONS: Detection and reporting of taeniosis in western Europe should be improved. The existence of T. solium tapeworm carriers, of suspected autochthonous cases of human cysticercosis and the lack of confirmation of porcine cysticercosis cases deserve further attention. Suspected cases of T. solium in pigs should be confirmed by molecular methods. Both taeniosis and human cysticercosis should be notifiable and surveillance in animals should be improved.

Narrowing of the Diagnostic Gap of Acute Gastroenteritis in Children 0-6 Years of Age Using a Combination of Classical and Molecular Techniques, Delivers Challenges in Syndromic Approach Diagnostics.

BACKGROUND: Twenty-five percent to 50% of acute gastroenteritis (AGE) cases remain etiologically undiagnosed. Our main aim was to determine the most appropriate list of enteric pathogens to be included in the daily diagnostics scheme of AGE, ensuring the lowest possible diagnostic gap. METHODS: Two hundred ninety seven children ≤6 years of age, admitted to hospital in Slovenia, October 2011 to October 2012, with AGE, and 88 ≤6 years old healthy children were included in the study. A broad spectrum of enteric pathogens was targeted with molecular methods, including 8 viruses, 6 bacteria and 2 parasites. RESULTS: At least one enteric pathogen was detected in 91.2% of cases with AGE and 27.3% of controls. Viruses were the most prevalent (82.5% and 15.9%), followed by bacteria (27.3% and 10.2%) and parasites (3.0% and 1.1%) in cases and controls, respectively. A high proportion (41.8%) of mixed infections was observed in the cases. For cases with undetermined etiology (8.8%), stool samples were analyzed with next generation sequencing, and a potential viral pathogen was detected in 17 additional samples (5.8%). CONCLUSIONS: Our study suggests that tests for rotaviruses, noroviruses genogroup II, adenoviruses 40/41, astroviruses, Campylobacter spp. and Salmonella sp. should be included in the initial diagnostic algorithm, which revealed the etiology in 83.5% of children tested. The use of molecular methods in diagnostics of gastroenteritis is preferable because of their high sensitivity, specificity, fast performance and the possibility of establishing the concentration of the target. The latter may be valuable for assessing the clinical significance of the detected enteric, particularly viral pathogens.

Multilocus genotyping of Giardia duodenalis (Lambl, 1859) from symptomatic human infections in Slovenia.

Giardiasis is a common gastrointestinal infection of humans and animals with a worldwide distribution. Eight genetic groups (known as assemblages A to H) are currently recognised within the species complex of Giardia duodenalis (Lambl, 1859), of which assemblages A and B are responsible for infection of humans and other mammalian hosts. Genotyping data on giardiasis are not available from Slovenia. In this work, we have characterised isolates of G. duodenalis from 85 human symptomatic cases collected during 2002-2013. Genomic DNAs were first tested by a real-time (rt) PCR assay and then by conventional PCR at three loci (beta-giardin, bg; triose phosphate isomerase, tpi; and glutamate dehydrogenase, gdh). We found that the threshold cycle (Ct) values in rt-PCR testing were higher for samples collected during 2002-2005 and that this was paralleled by a low amplification rate in conventional PCR (6 of 32, i.e. 19%). In contrast, lower Ct values and higher amplification rate (45 of 53; 85%) were observed for samples collected during 2006-2013, suggesting an adverse effect of prolonged freezing of stools. Assemblages A and B were found with an almost identical frequency in the 51 genotyped samples. In agreement with previous studies, sequences from assemblage B isolates were characterised by larger genetic variability and by the presence of heterogeneous positions, which made assignment to specific genotypes difficult. Less variability was observed in sequences from assemblage A isolates, which belonged to the human-specific subassemblage AII. These data showed that the genotypes of G. duodenalis that circulate in humans in Slovenia are similar to those previously identified in Europe.

A real-time assemblage-specific PCR assay for the detection of Giardia duodenalis assemblages A, B and E in fecal samples.

Giardiosis is a common gastrointestinal infection caused by the flagellate Giardia duodenalis, and affects both humans and animals, worldwide. Animals are infected with both zoonotic and host-specific G. duodenalis assemblages, and their role in the transmission of the infection to humans has been a subject of intense research and debate. Conventional PCR assays are appropriate to determine G. duodenalis assemblages, but lack sensitivity for the detection of mixed infections. Previous surveys demonstrated the occurrence of mixed infections with G. duodenalis assemblage A and B in humans, and with assemblages A and E in cattle, but are likely to be underestimated. In this study, we designed a set of assemblage-specific primers by exploiting sequence variability in homologous genes from assemblages A, B and E. Primers were designed to amplify fragments of different size that generated different melting curves from each assemblage in real-time PCR (rt-PCR) experiments. The assay has been tested on a large panel of human and farm animal isolates, and shown to possess high specificity (no cross reactions observed) and sensitivity (detection limit close to 20 copies). Therefore, this assay can be useful to detect zoonotic and host-specific G. duodenalis assemblages in fecal samples from farm animals, particularly when a large number of samples is to be tested.

Trichomoniasis: a brief review of diagnostic methods and our experience with real-time PCR for detecting infection.

Trichomoniasis is the most common non-viral sexually transmitted infection, and it is caused by the protozoan flagellate Trichomonas vaginalis. Although highly prevalent in sexually active women, it has long been overlooked in other groups of potentially infected people. Recently, studies have shown that trichomoniasis increases the risk of infection with human immunodeficiency virus and can cause adverse outcomes of pregnancy, which has increased interest in T. vaginalis and increased the need for highly sensitive diagnostic tests. This article summarizes the diagnostic methods most commonly used in the diagnosis of trichomoniasis, including the most sensitive and specific nucleic acid amplification tests. It also presents the results of our study comparing the performance of wet mount microscopy and culture to real-time PCR for detecting the parasite.

Evidence of human neurocysticercosis in Slovenia.

To assess the prevalence of Taenia solium cysticercosis in patients with neurological disorders in Slovenia, serum/cerebrospinal fluid (CSF) samples from 348 suspected patients were collected between the beginning of January 2001 and the end of December 2012 and analysed serologically for the presence of anti-T. solium IgG antibodies. Of 20 patients whose samples tested positive or equivocal by enzyme-linked immunosorbent assay (ELISA), samples of 7 patients were confirmed positive by Western blot (WB). The overall seroprevalence rate of T. solium infection in patients with neurological disorders included in the study was 2.0%. Serological results of positive patients corresponded to clinical and/or imaging findings concerning their brain cysts. Based on their personal data, it was ascertained that neurocysticercosis (NCC) positive patients had immigrated or came to Slovenia from the former Yugoslav republics. Since the disease is believed not to be endemic in Slovenia we assume that all of the NCC-positive patients had acquired the infection before immigration to Slovenia or visiting or being visited by their relatives infected with an adult T. solium parasite. The present results represent the first insight into the prevalence of NCC in patients with neurological disorders in Slovenia.

Serological evidence for human cystic echinococcosis in Slovenia.

BACKGROUND: Cystic echinococcosis (CE) is caused by the larva of tapeworm Echinococcus granulosus. Dogs and other canids are the primary definitive hosts for this parasite. CE may develop after accidental ingestion of tapeworm eggs, excreted with the feces of these animals. In the intestine, the larvae released from the eggs are nested in the liver, lungs or other organs of livestock as intermediate hosts and humans as aberrant hosts. The aim of this study was to examine serologically whether some of the patients in Slovenia, suspected of CE by imaging findings in the liver or lungs had been infected with the larva of Echinococcus granulosus. METHODS: Between January 1, 2002 and the end of December 2006, 1323 patients suspected of having echinococcosis were screened serologically by indirect haemagglutination assay (IHA). For confirmation and differentiation of Echinococcus spp. infection, the sera of IHA-positive patients were then retested by western blot (WB). RESULTS: Out of 127 IHA-positive sera, 34 sera were confirmed by WB and considered specific for CE. Of 34 sera of CE-positive patients sera, 32 corresponded to the characteristic imaging findings of a liver cysts and 2 to those of lung cysts. The mean age of CE-positive patients was 58.3 years. No significant differences were found between the CE-positive patients in regard to their sex. CONCLUSION: In the study, it was found out that CE was mostly spread in the same area of Slovenia as in the past, but its prevalence decreased from 4.8 per 105 inhabitants in the period 1956-1968 to 1.7 per 105 inhabitants in the period 2002-2006. In spite of the decreased prevalence of CE in the last years, it is suggested that clinicians and public health authorities, especially in the eastern parts of Slovenia where the most CE patients come from, should pay greater attention to this disease in the future.