Chronic intestinal schistosomiasis caused by co-infection with Schistosoma intercalatum and Schistosoma mansoni.

The Grand Round concerns a 24-year-old man from Zimbabwe who was studying and living in Poland. The patient had been complaining of abdominal pain, fatigue, alternating diarrhoea and constipation, and presence of blood in his stool for 3 years. The patient had the following diagnostic tests: colonoscopy, CT scan, histopathology, and parasitological and molecular tests. Results of the examinations showed that the cause of the patient's complaints was chronic intestinal schistosomiasis due to the co-infection with Schistosoma intercalatum and Schistosoma mansoni. The patient had two cycles of praziquantel therapy (Biltricide) and responded well to the treatment. In the Grand Round, we describe full diagnostics as well as clinical and therapeutic management in the patient with S intercalatum and S mansoni co-infection. This case allows us to draw attention to cases of forgotten chronic tropical diseases (including rare ones) in patients from regions with a high endemic index staying in non-endemic regions of the world for a long time. Co-infection with S intercalatum and S mansoni should be considered as a very rare clinical case.

Successive Acanthamoeba Corneal Isolates Identified in Poland Monitored in Terms of In Vitro Dynamics.

BACKGROUND: Amoebae of the genus Acanthamoeba cause a sight-threatening infection called Acanthamoeba keratitis. It is considered a rare disease in humans but poses an increasing threat to public health worldwide, including in Poland. We present successive isolates from serious keratitis preliminary examined in terms of the identification and monitoring of, among others, the in vitro dynamics of the detected strains. METHODS: Clinical and combined laboratory methods were applied; causative agents of the keratitis were identified at the cellular and molecular levels; isolates were cultivated in an axenic liquid medium and regularly monitored. RESULTS: In a phase-contrast microscope, Acanthamoeba sp. cysts and live trophozoites from corneal samples and in vitro cultures were assessed on the cellular level. Some isolates that were tested at the molecular level were found to correspond to A. mauritanensis, A. culbertsoni, A. castellanii, genotype T4. There was variability in the amoebic strain dynamics; high viability was expressed as trofozoites' long duration ability to intense multiply. CONCLUSIONS: Some strains from keratitis under diagnosis verification and dynamics assessment showed enough adaptive capability to grow in an axenic medium, allowing them to exhibit significant thermal tolerance. In vitro monitoring that was suitable for verifying in vivo examinations, in particular, was useful to detect the strong viability and pathogenic potential of successive Acanthamoeba strains with a long duration of high dynamics.

Misdiagnosis and inappropriate treatment of cutaneous leishmaniasis: a case report.

BACKGROUND: Leishmaniasis is a widespread disease in tropical and subtropical countries, except for Australia and Oceania. In Poland, tourists, migrants and travellers from leishmaniasis-endemic countries may carry Leishmania. CASE PRESENTATION: We present a case of undiagnosed cutaneous leishmaniasis in a patient who received many weeks of inadequate antibiotic treatment. Ulceration in the right submandibular region was thought to be a purulent complication after laser surgery. Six weeks before the ulcer developed, the patient had visited the jungle (Guatemala). Cutaneous leishmaniasis was finally diagnosed after nine months based on a proper history and a polymerase chain reaction (PCR) assay. Treatment with antimony derivatives was administered. After three months, the ulcer healed but left a scar. CONCLUSION: A lack of knowledge about tropical diseases among doctors and an incomplete medical history were the reasons for many weeks of erroneous treatment of cutaneous leishmaniasis with antibiotics. This is the first reported case of cutaneous leishmaniasis misdiagnosed as a complication after an aesthetic medical procedure.

Rare Occurrence of Blastocystis in Pet Animals and Their Owners in the Pomeranian Voivodeship in Poland in the Light of Literature Data.

Blastocystis is an intestinal microeukaryote with ambiguous pathogenicity, commonly detected in human feces worldwide. It comprises at least 28 genetically diverse subtypes (STs), 12 of which also occur in a wide range of animal species, giving rise to suspicion of zoonotic transmission. To investigate this, we conducted a molecular study of 145 stool samples of pet animals, and 67 of their owners, living in an urban area in Poland. Blastocystis was detected in only three (2.1%) animal samples (of two bearded agamas and a leopard gecko), while all dogs, cats, and pet rodents were Blastocystis-negative. Blastocystis was also present in three (4.5%) owners of animals, but they were cat owners, not reptile owners, and the subtypes identified in them differed significantly from those of reptiles. Additionally, the frequency of Blastocystis in different groups of dogs (depending on how they were kept) was analyzed. This work is the first to find Blastocystis in pet reptiles, and we encourage further investigation of Blastocystis in this poorly examined group of animals, as well as continued study on the transmission of this microorganism between humans and animals.

Concurrent subcutaneous and ocular infections with Dirofilaria repens in a Polish patient: a case report in the light of epidemiological data.

Dirofilariasis is an emerging zoonosis caused by nematodes of the genus Dirofilaria, most often D. repens and D. immitis. The main final hosts and reservoirs of pathogens are dogs. The intermediate hosts and vectors of infection are female mosquitoes (Culicidae). Human is an accidental host in which the parasite does not usually mature. Over the past 20 years, the range of Dirofilaria spp. in Europe has expanded. We present an unusual case of multifocal dirofilariasis of mixed subcutaneous-ocular course caused by D. repens in a 52-year-old Polish patient who was probably infected in Spain or Croatia, where she stayed one year before the onset of symptoms. Surgical removal of the nematodes followed by treatment with Ivermectin in a single dose of 1200 µg and Doxycycline 200 mg daily for 7 days resulted in complete recovery. We believe that all cases of human dirofilariasis, especially in countries where the disease is not frequent at present, should be registered for epidemiological purposes. Moreover, due to the widening of the range of D. repens and D. immitis occurrence and the possibility of atypical courses of infection with both nematodes, diagnostics should include the species identification of the parasite.

Molecular Characterization of Blastocystis from Animals and Their Caregivers at the Gdansk Zoo (Poland) and the Assessment of Zoonotic Transmission.

Blastocystis is a highly genetically diverse gut protist commonly found in humans and various animals. The role of animals in human infection is only partly understood. The aim of this study was to determine the host specificity and possibility of zoonotic transmission of this microorganism. Subtypes of Blastocystis isolated from 201 zoo animals and their 35 caregivers were identified by sequencing of the SSU rRNA gene. Blastocystis was found in 26.86% of animal and 17.14% of human samples. Both mammalian (ST1-ST3, ST5, ST8, ST10, ST13, ST14) and non-mammalian subtypes were detected. Of the subtypes found in non-human primates (ST1, ST2, ST3, and ST13), two subtypes (ST1 and ST3) were also detected in humans. The presence of identical ST1 sequences in three monkeys and their caregiver indicates the possibility of direct transmission of Blastocystis between these animals and humans. Detection of ST5 only in wild boars and peccaries, ST8 only in Marsupial, ST10 and ST14 only in Bovidae, and non-mammalian subtypes in reptiles suggests higher host specificity for these subtypes, and indicates that their transmission between animals and humans is unlikely. Additionally, this was probably the first time that ST5 was found in peccaries, ST2 in patas monkeys, and ST8 in red kangaroos.

First Report on the Occurrence and Subtypes of Blastocystis in Pigs in Poland Using Sequence-Tagged-Site PCR and Barcode Region Sequencing.

Blastocystis is an enteric microorganism commonly found in humans and animals worldwide. Its pathogenic role in humans and transmission patterns has not been fully explained. However, nine subtypes (ST1-8, ST12) are considered as potentially zoonotic. Studies from various regions of the world show that pigs are mainly infected with ST5. Although pigs are important farmed animals in Poland, the question of Blastocystis infection in these animals has not yet been investigated. Herein, 149 pig stool samples from 10 Polish pig farms were analyzed using sequence-tagged-site PCR and barcode region sequencing. The percentage of samples in which Blastocystis was identified using each method separately was similar: 38.25% and 37.58%, respectively. However, the percentage of positive results obtained by combining both methods was 46.97%, which means that, depending on the method used, the number of undetected samples varied between 8.72% and 9.39%. This shows the methodological limitations of up-to-date molecular approaches commonly used in Blastocystis research. A moderate infection rate (44.4-50%) observed in different pig age groups with a vital predominance of ST5 (94.28%) in every age group shows that pigs are a likely natural host of ST5. A small percentage of mixed infections, namely ST5/ST1 (5.26%), ST5/ST3 (1.75%), and ST3/ST1 (1.75%), was observed only in animals of older age, suggesting that ST3 and ST1 can be acquired by pigs during contact with humans. This study provides the first data on the prevalence and Blastocystis subtypes (STs) distribution in pigs in Poland. The results also highlight the need for the development of new methods capable of detecting highly genetically diverse Blastocystis isolates and mixed infections.

First case of human anisakiasis in Poland.

Consumption of raw or inadequately processed marine fish may result in anisakidosis - a zoonotic disease caused by larvae of the parasitic nematodes of the family Anisakidae (anisakiasis when caused by members of the genus Anisakis (Nematoda: Anisakidae)), commonly found in a variety of marine fish species all over the world. Most cases of anisakidosis have been detected in the residents of Japan and South Korea, which results from the tradition of eating raw and semi-raw fish dishes. However, the disease is now increasingly often diagnosed in other parts of the world, including Europe (mainly in Spain and Italy). In Poland, no cases of human infection with anisakid nematodes have been detected so far. In this study, we report the first case of gastric anisakiasis in Poland, in a 59-year-old female patient, after eating raw Atlantic salmon (Salmo salar). The parasite was identified as the third-stage larva of Anisakis simplex sensu stricto on the basis of morphology and molecular analysis. The larva was still alive and causing pain until it was removed, which occurred more than 5 weeks after infection. The described case prove that anisakiasis should be considered as a potential cause of gastrointestinal tract ailments following the consumption of seafood in countries where no cases of this zoonosis have been reported to date.

First evidence of the presence of Anisakis simplex in Crangon crangon and Contracaecum osculatum in Gammarus sp. by in situ examination of the stomach contents of cod (Gadus morhua) from the southern Baltic Sea.

Cod (Gadus morhua), an important fish species in the Baltic Sea, is the paratenic host for many parasite species, including the zoonotic nematodes, Anisakis sp. and Contracaecum osculatum. We aimed to identify which invertebrate species (found in situ in the fish stomach) are responsible for infection of cod with zoonotic nematodes. We found that Crangon crangon and Gammarus sp., both invertebrate prey species of cod, were infected with Anisakis simplex and C. osculatum, respectively. These host-parasite systems are reported here for the first time, implicating C. crangon and Gammarus sp. as sources of infection of Baltic cod with zoonotic nematodes.

Blastocystis subtypes isolated from travelers and non-travelers from the north of Poland - A single center study.

Blastocystis is a common, enteric protist of humans and animals with a worldwide distribution and unclear clinical relevance. Nine out of 17 genetically diverse subtypes occur in humans. We analysed the distribution of Blastocystis subtypes and the intensity of invasion in relation to the gastrointestinal tract disorders and travels to different continents. 122 Blastocystis stool cultures were subtyped via polymerase chain reaction (PCR) with seven pairs of subtype-specific, sequence-tagged-site (STS) primers. Five subtypes of Blastocystis were detected: ST3 (59%), ST2 (19.7%), ST1 (13.1%), ST6 (3.3%), ST7 (3.3%), and two mixed infections with ST1/ST3 (1.6%). ST1 was detected exclusively in travelers to hot climate zones and ST2 was found more frequently in people visiting other continents compared to those who never left Poland. We found no correlation between gastrointestinal tract disorders, Blastocystis STs, and parasite load. There was no age predisposition to the Blastocystis infection. We established the distribution of Blastocystis STs among Poles traveling to different continents and never leaving Poland. Our study sheds more light on the problem of importing Blastocystis infection. It shows that certain subtypes detected in Europe can be imported due to travel or migration. Collecting data on the travel history of the surveyed persons is necessary to clarify this matter.

First detection of Echinococcus multilocularis in environmental water sources in endemic areas using capsule filtration and molecular detection methods.

Water is one of the possible transmission routes for water- and foodborne parasites to humans. Echinococcus multilocularis is a parasite, which causes alveolar echinococcosis (AE). Nevertheless, no environmental studies have been performed as yet to confirm the occurrence of E. multilocularis in water supplies. Accordingly, 105 water samples of 50 L volume were collected from surface waters (lakes, rivers, canals) and wells in the Warmia-Masuria Province (Echinococcus endemic area) and Pomerania Province (Echinococcus non-endemic area), Poland. The water was filtered and subsequently analysed with nested PCR and real-time PCR. E. multilocularis DNA was found in two (1.9%) samples, which originated from two lakes localised in the Warmia-Masuria Province. Sequencing of the positive samples confirmed that the PCR products were fragments of the E. multilocularis mitochondrial 12S rRNA gene. This is the first investigation describing E. multilocularis detection in environmental water samples, using molecular diagnostic tools. The results indicate that water could be considered as a potential source of E. multilocularis infections in humans and animals, in endemic areas.

How effective is freezing at killing Anisakis simplex, Pseudoterranova krabbei, and P. decipiens larvae? An experimental evaluation of time-temperature conditions.

The consumption of raw or inadequately cooked marine fish can lead to several disorders caused by the ingestion of viable anisakid nematodes. Although anisakid larvae can be killed by subzero temperatures, making freezing an important control measure for this potential health hazard, these parasites can survive freezing under some conditions. Therefore, the aim of the present study was to experimentally evaluate the time-temperature conditions needed to kill Anisakis simplex and Pseudoterranova spp. The effectiveness of freezing was tested on two species of fish: cod, Gadus morhua from the North Atlantic, and herring, Clupea harengus membras from the southern Baltic Sea. Samples, which comprised skinless fillets of cod (n = 40) with visible parasites and whole herring (n = 240), were separately frozen at - 15, - 18, or - 20 °C for 24 h, or at - 20 °C for 48 h in the single-compressor freezer and at - 20, - 25, or - 35 °C for 24 h in the double-compressor freezer. After thawing, parasites were stained with malachite green and examined under the microscope for viability. All A. simplex and Pseudoterranova spp. larvae in cod fillets died at a temperature of - 15 °C or lower. However, freezing did not kill all the A. simplex larvae in whole herring: spontaneous movement of these parasites was observed in samples stored in the single-compressor freezer at - 15, - 18, and - 20 °C over 24 h. Our results demonstrate that the freezing procedure must consider both the capability of the freezing device and the nature of the fish product to ensure consumer safety.

The First Case of Elbow Bursitis Caused by Dirofilaria repens in Humans.

Our case report involves a 28-year-old man who was diagnosed with left elbow bursitis. After thorough macroscopic and microscopic examinations and serological and molecular tests, it was found that the inflammation had been caused by a Dirofilaria repens infection. This case report is the world's first documented description.

Ultrasound images in hepatic alveolar echinococcosis and clinical stage of the disease.

PURPOSE: Hepatic alveolar echinococcosis (AE) is a parasitic disease caused by the larval stage of the tapeworm Echinococcus multilocularis. Ultrasonography is the method of choice in the initial diagnosis of AE. The aim of the study is to present the most frequent sonomorphological patterns of lesions in hepatic AE based on the analysis of ultrasound findings in patients treated for AE at the University Centre of Maritime and Tropical Medicine (UCMMiT; Gdynia, Poland), and to establish whether there is a relationship between the clinical stage of AE and the occurrence of a specific sonomorphological pattern of hepatic lesions. PATIENTS AND METHODS: We analysed the results of ultrasound examinations of 58 patients hospitalized in the UCMMiT with probable or certain diagnosis of AE. Liver lesions were assessed according to the classification developed by researchers from the University Hospital in Ulm (Germany). Statistical analysis was based on the relationship between the occurrence of a specific sonomorphological pattern of hepatic lesions and the clinical stage of AE. RESULTS: The most frequently observed patterns of AE lesions in the liver were the hailstorm and the pseudocystic patterns. There was no correlation between the clinical stage of the disease and the ultrasonographic appearance of lesions. There was no statistically significant relationship between the more frequent occurrences of specific ultrasonographic patterns of lesions in the liver and radical or non-radical surgery. CONCLUSIONS: The ultrasonographic appearance of the lesion in liver AE cannot determine the therapeutic management. Treatment plan should be established based on the PMN classification.

The usefulness of commercially available serological tests in the diagnosis and monitoring of treatment in patients with alveolar echinococcosis.

AIM OF THE STUDY: To assess the clinical usefulness of serological tests in the diagnosis and monitoring of treatment of patients with alveolar echinococcosis (AE). MATERIAL AND METHODS: The results of serological tests, i.e. Echinococcus multilocularis ELISA (Bordier Affinity Products) and Echinococcus Western Blot IgG (LDBIO Diagnostic), of 66 patients were analysed. Duration of follow-up was two years after diagnosis. In the second phase of the study 11 sera obtained from the patients undergoing surgical treatment, in whom the results of Echinococcus Western Blot IgG assay were still positive, were additionally tested with Anti-Echinococcus EUROLINE-WB (IgG) assay. RESULTS: Statistically significant negativization of the Echinococcus multilocularis ELISA test was observed in the group of patients who underwent radical surgery or liver transplantation. Negativization of Echinococcus Western Blot IgG assay results was observed in some patients, among both those who received conservative treatment and those who underwent surgery, but no statistically significant differences were found between treatment groups. In 54.5% of cases the Anti-Echinococcus EUROLINE-WB (IgG) test result was negative when the results of the Echinococcus Western Blot IgG assay were still positive. CONCLUSIONS: Echinococcus multilocularis ELISA proved to be useful in assessing the activity of AE in a group of patients who underwent radical surgery or liver transplantation. The results of our study suggest that Anti-Echinococcus EUROLINE-WB (IgG) is a more dynamic test, which at the time of disappearance of AE activity becomes negative earlier.

Detection of Giardia intestinalis in water samples collected from natural water reservoirs and wells in northern and north-eastern Poland using LAMP, real-time PCR and nested PCR.

Giardia intestinalis is a protozoan parasite, transmitted to humans and animals by the faecal-oral route, mainly through contaminated water and food. Knowledge about the distribution of this parasite in surface water in Poland is fragmentary and incomplete. Accordingly, 36 environmental water samples taken from surface water reservoirs and wells were collected in Pomerania and Warmia-Masuria provinces, Poland. The 50 L samples were filtered and subsequently analysed with three molecular detection methods: loop-mediated isothermal amplification (LAMP), real-time polymerase chain reaction (real-time PCR) and nested PCR. Of the samples examined, Giardia DNA was found in 15 (42%) samples with the use of LAMP; in 12 (33%) of these samples, Giardia DNA from this parasite was also detected using real-time PCR; and in 9 (25%) using nested PCR. Sequencing of selected positive samples confirmed that the PCR products were fragments of the Giardia intestinalis small subunit rRNA gene. Genotyping using multiplex real-time PCR indicated the presence of assemblages A and B, with the latter predominating. The results indicate that surface water in Poland, as well as water taken from surface wells, may be a source of Giardia strains which are potentially pathogenic for humans. It was also demonstrated that LAMP assay is more sensitive than the other two molecular assays.

The first detection of Toxoplasma gondii DNA in environmental air samples using gelatine filters, real-time PCR and loop-mediated isothermal (LAMP) assays: qualitative and quantitative analysis.

Toxoplasma gondii infections are acquired through the ingestion of oocysts present in the environment. However, there is no data about their occurrence in the air or about airborne transmission of these infections. In the present paper, we report on the identification of T. gondii using rapid molecular detection methods, supported by microscopic analysis, in environmental air samples. A total of 71 samples were collected, using gelatine filters, from kitchen gardens, recreational areas and sandpits located in northern and north-eastern Poland. Material recovered from the filters was analysed using real-time PCR and loop-mediated isothermal assays targeting the T. gondii B1 gene. Toxoplasma gondii DNA was found in two samples, as confirmed by both molecular assays. Genotyping at the SAG2 locus showed Toxoplasma SAG2 type I. Moreover, the presence of T. gondii oocysts was confirmed in one of the positive samples with the use of microscopy. The results showed that T. gondii may be present in environmental air samples and that respiratory tract infections may play a role in the high prevalence of toxoplasmosis in humans and animals. To the best of our knowledge, this is the first epidemiological evidence that oro-fecal and foodborne toxoplasmosis may be traceable to an airborne respiratory origin and that this may represent a new, previously unknown transmission route for this disease.

Detection of Echinococcus multilocularis DNA in fruit, vegetable, and mushroom samples collected in the non-endemic territory of the Pomerania province and comparison of the results with data from rural areas of the neighbouring highly endemic Warmia-Masuria province, Poland.

Echinococcus multilocularis is a tapeworm that may cause alveolar echinococcosis (AE), one of the most dangerous parasitic zoonoses. As in the case of some foodborne diseases, unwashed fruits and vegetables contaminated with eggs of E. multilocularis may serve as an important transmission route for this parasite. The aim of this study was to investigate the presence of E. multilocularis DNA in fruit, vegetables, and mushrooms in rural areas of the Pomerania province, Poland (non-endemic territory). In total, 104 environmental fruit, vegetable, and mushroom samples collected in forests, plantations, and kitchen gardens were analysed using nested PCR based on the mitochondrial 12S rRNA gene. E. multilocularis DNA was detected in 6.7 % of the samples tested, which indicated that the environment of the Pomerania province is contaminated with this parasite, creating a potential risk for humans. Therefore, fresh fruit, vegetables, and mushrooms should be washed before consumption. Additionally, the results showed that the level of contamination is significantly lower than in the highly endemic Warmia-Masuria province. The differences in the occurrence of E. multilocularis in the environment of these neighbouring provinces appears to be connected with the general epidemiological situation of these two regions, but further study is required for an exact explanation.

Cryptosporidium meleagridis infection: the first report in Poland of its occurrence in an HIV-positive woman

Cryptosporidium is an opportunistic protozoan parasite that can cause severe diarrhoea in immunocompromised patients. The transmission of this pathogen in humans and animals is not fully understood. C. meleagridis, originally described in birds, is the only Cryptosporidium species known to naturally infect mammalian and avian species. This study documents the first detection of C. meleagridis in an HIV-infected woman in Poland.

Fresh fruits, vegetables and mushrooms as transmission vehicles for Echinococcus multilocularis in highly endemic areas of Poland: reply to concerns.

Echinococcus multilocularis is a tapeworm that may cause alveolar echinococcosis (AE), one of the most dangerous parasitic zoonoses. As in the case of other foodborne diseases, unwashed fruits and vegetables, contaminated with dispersed forms of E. multilocularis, may serve as an important transmission route for this parasite. In this article, we reply to the incorrect interpretation of results of our study concerning the detection of E. multilocularis DNA in fresh fruit, vegetable and mushroom samples collected from the highly endemic areas of the Warmia-Masuria Province, Poland, to dispel any doubts. The accusations formulated by the commentators concerning our paper are unfounded; moreover, these commentators demand information which was beyond the purview of our study. Making generalisations and drawing far-reaching conclusions from our work is also unjustified. The majority of positive samples were found in only a few hyperendemic communities; this information corresponds with the highest number of both infected foxes and AE cases in humans recorded in this area. Our findings indicate that E. multilocularis is present in the environment and may create a potential risk for the inhabitants. These people should simply be informed to wash fruits and vegetables before eating. No additional far-reaching conclusions should be drawn from our data. We believe these commentators needlessly misinterpreted our results and disseminated misleading information. Nevertheless, we would like to encourage any readers simply to contact us if any aspects of our study are unclear.