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Rodents are known to be reservoirs of Toxoplasma gondii and keep the parasite circulation in the environment. We conducted biomonitoring to assess the role of sylvatic rodents in maintaining T. gondii and to analyse the prevalence and seroprevalence of the parasite in seven wild rodent species. Rodents were collected in our study sites (woodland and open grasslands) located in northeastern Poland and dissected. We collected brain, spleen, blood and serum samples. We applied both molecular (PCR assay, nested-PCR assay) and serological (ELISA and agglutination tests) methods to indicate the best approach for application in the biomonitoring of T. gondii in small mammals. We screened samples from 95 individuals sing PCR assays and found no T. gondii DNA. The agglutination test showed no signal. We found antibodies against T. gondii in 6 sera samples out of 74 analysed (seroprevalence = 8.11% [4.0-17.1]). Our results confirm that rodents participate in the life cycle of T. gondii as reservoirs of this parasite in the sylvatic environment. However, biomonitoring should be performed with the ELISA tests to search for T. gondii antigens, rather than a molecular approach only.

INTRODUCTION Malaria remains a diagnostic and therapeutic challenge in many endemic regions of sub-Saharan Africa. It is one of the most important causes of morbidity and mortality, especially in children

Toxoplasma gondii infection is becoming increasing problem in China but there is no data concerning contamination of vegetables intended for consumption with this parasite. The aim of the present study was to investigate fresh vegetables originated from open markets located in the Xining City, the Qinghai-Tibet Plateau (QTP), P.R. China for their contamination with T. gondii. A total of 279 fresh vegetable samples were collected and analysed using real-time PCR assay targeting B1 gene and multilocus genotyping. T. gondii DNA was found in 10 (3.6%) samples tested; eight of them represented T. gondii type I and remaining two T. gondii type II. The approximate level of contamination of positive vegetables samples, estimated based on quantitative real-time PCR (qPCR), ranged between less than one and 27000 T. gondii oocysts per sample, with majority not exceeding several oocysts per sample. The results of the study confirmed that T. gondii is present in vegetables offered in open markets in the Qinghai province, P.R. China; eating them unwashed and raw may therefore pose a threat to consumers. This is the first investigation describing T. gondii detection in fresh vegetables intended for consumption collected from the territory of P.R. China using sensitive molecular tools.

Malaria remains a diagnostic and therapeutic challenge in many endemic regions of sub-Saharan Africa. It is one of the most important causes of morbidity and mortality, especially in children5 years.The objective of the study was to assess the prevalence ofThe study was conducted during the dry and rainy seasons in 2018 and involved a group of 540 symptomatic BaAka Pygmies, patients of both genders, aged 1-75-years-old. Two diagnostic methods for detecting Plasmodium in the bloodstream were used: RDTs targeting HRP2-protein specific forOnly 40.5% of symptomatic patients tested with RDTs forThe study suggests the need for introducing accurate diagnostic methods for the diagnosis of malaria and the revision of malaria treatment protocols. Assessment of the Pfhrp2/Pfhrp3 deletions is necessary for evaluating malaria epidemiology in Central Africa.

According to the World Health Organization 94% of global malaria cases and 94% of global malaria deaths have been reported from Africa. Unfortunately, it is difficult to determine the exact prevalence of disease in some African countries due to a large number of asymptomatic cases. The aim of this study was to assess the prevalence of malaria infections in seemingly healthy children living in the Central African Republic (CAR). CareStartTM Malaria HRP2 rapid diagnostic test (RDT) targeting Plasmodium falciparum was used to test a group of 500 asymptomatic children aged 1-15 years old (330 settled Bantu and 170 semi-nomadic BaAka Pygmies) inhabiting the villages in the Dzanga Sangha region (south-west CAR) in March 2020. In total, 32.4% of asymptomatic Bantu and 40.6% of asymptomatic Pygmy children had a positive result of malaria RDT. Our findings allowed us to demonstrate the high prevalence of asymptomatic malaria infections in south-west CAR. RDTs seem to be a useful tool for the detection of Plasmodium falciparum in areas with limited possibilities of using other diagnostic methods, such as light microscopy and molecular biology.

Background: Malaria remains a diagnostic and therapeutic challenge in many endemic regions of sub-Saharan Africa. The objective of the study was to assess the prevalence of Plasmodium spp. in BaAka Pygmies with clinical symptoms of malaria, inhabitants of the rural Dzanga Sangha region in Central African Republic. Additionally, the research aimed to define the percentage distribution of infections caused by other than P. falciparum species in order to assess the need for diversification of malaria treatment protocols in the region of Central Africa. Material and methods: The study was conducted during dry and rainy season in 2018 and involved a group of 540 symptomatic BaAka Pygmies, patients of both sexes, aged 1–75 years old. Two diagnostic methods for detecting Plasmodium in the bloodstream were used: RDTs targeting HRP2–protein specific for P. falciparum and PCR assays aimed at detecting P. falciparum, P. vivax, P. ovale, P. malariae species. Results: Only 40.5% of symptomatic patients tested with RDTs for P. falciparum infections were positive. While molecular tests (PCR) confirmed P. falciparum in 94.8% of the samples and also revealed the genetic material of P. malariae (11.1%), P. ovale (9.8%), and P. vivax (0.7%). BaAka Pygmies aged Conclusions: The presented study suggest the need for introducing more accurate diagnostic methods for the diagnosis of malaria and the revision of malaria treatment protocols. The assessment of the Pfhrp2/Pfhrp3 deletions is necessary for evaluating malaria epidemiology in Central Africa.

According to the World Health Organization 94% of global malaria cases and 94% of global malaria deaths have been reported from Africa. Unfortunately, it is difficult to determine the exact prevalence of disease in some African countries due to a large number of asymptomatic cases. The aim of this study was to assess the prevalence of malaria infections in seemingly healthy children living in the Central African Republic (CAR). CareStartTM Malaria HRP2 Pf Ag RDT targeting Plasmodium falciparum was used to test a group of 500 asymptomatic children aged 1-15 years old (330 settled Bantu and 170 semi-nomadic BaAka Pygmies) inhabiting the villages in the Dzanga Sangha region (south-west CAR) in March 2020. 32.4% of asymptomatic Bantu and 40.6% of asymptomatic Pygmy children had a positive result of malaria RDT. The mean age of the study participants with RDT (+) was 8.7 in Bantu and 7.0 years in Pygmies; the mean body temperature was 36.8oC in both groups; the mean haemoglobin level was 10.6 g/dL and 10.1 g/dL, respectively. Our findings allowed us to demonstrate the high prevalence of asymptomatic malaria infections in south-west CAR. RDTs seem to be a useful tool for the detection of Plasmodium falciparum in areas with limited possibilities of using other diagnostic methods, such as light microscopy and molecular biology.

SUMMARYToxoplasma gondiiinfections 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 ofT. gondiiusing 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 theT. gondiiB1 gene.Toxoplasma gondiiDNA was found in two samples, as confirmed by both molecular assays. Genotyping at the SAG2 locus showedToxoplasmaSAG2 type I. Moreover, the presence ofT. gondiioocysts was confirmed in one of the positive samples with the use of microscopy. The results showed thatT. gondiimay 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.

It is estimated that faecal-orally transmitted diseases are common in Afghanistan, as a consequence of poor hygienic standards of life and widespread contamination of water and food with both human and animal faeces. However, there is little information in the literature concerning infections caused by intestinal parasites in the Afghan population. In this study, we report the occurrence of Giardia intestinalis assemblages (A and B) in formalin-fixed stool samples collected from 245 Afghan schoolchildren living in Ghazni Province in eastern Afghanistan. Detection of the parasite’s DNA and genotyping was performed using real-time PCR, specific to the β-giardin gene of G. intestinalis. Positive results were recorded in 52 (21.2%) samples. Genotyping was successful in 39 faecal samples and showed the predominance of assemblage B of G. intestinalis in this population (15 assemblage A and 24 assemblage B). Co-infection with both genotypes A and B was detected in four samples. Additionally, we evaluated the effect of 10% buffered formalin fixative on the detection of G. intestinalis DNA using real-time PCR and nested PCR characterised by different lengths of PCR products (74 and 479 bp, respectively). The human faeces containing the Giardia cysts were tested for 16 weeks. Amplification of G. intestinalis DNA with real-time PCR was possible up to 6 weeks of preservation of stool sample in formalin, compared to only 2 weeks with nested PCR. This suggests that real-time PCR is a more suitable tool in cases where stool samples have to be kept in formalin for longer periods of time.

Various species of amoebas belonging to the genus Acanthamoeba are widely distributed in many parts of the world. Some strains of these protozoans may exist as parasites and pose risks to human health as causative agents of serious human diseases. Currently in China there is a lack of information about the distribution of Acanthamoeba strains in the environment. Accordingly, 261 environmental water samples taken from rivers, sewage, and pharmaceutical factory drains were collected in Qinghai Province, China. The material was filtered and then analysed with both LAMP and PCR assays. Of the samples examined, Acanthamoeba DNA was found in 32 (14.68%) samples with the use of LAMP; in 13 of these samples, DNA from this amoeba was also detected using PCR. Sequencing of selected positive samples confirmed that the PCR products were fragments of the Acanthamoeba 18S rRNA gene and that isolates represent the T4 genotype, known as the most common strain related to AK cases. The results indicate that surface water, as well as water taken from sewage and pharmaceutical drains, may be a source of acanthamoebic strains potentially pathogenic for humans in China. It has been also demonstrated that LAMP assays is more sensitive than PCR and can be regarded as useful tool for screening the environment for Acanthamoeba spp.

Background. The prevalence rates of food- and waterborne parasitic infections in Afghanistan are unknown. Cases of invasive diseases found in Afghans are rarely laboratory-confirmed. Objectives . The aim of the study was to present the current status of intestinal parasitic infections in Afghan inhabitants on the example of patients hospitalized in two healthcare facilities in eastern Afghanistan. Material and methods . Fecal samples were collected from 548 patients (children aged 1–17 years and adults) with internal complaints, treated in Ghazni Provincial Hospital (Afghan civilian medical center, Ghazni province, 180 south-west of Kabul) and in Bagram Korean Hospital (Korean military medical center for Afghan patients, Parwan province, 60 km north of Kabul) between 2013 and 2014. One to three stool specimens from Afghan patients were fixed in 10% formalin, transported to the Military Institute of Medicine in Poland and tested by light microscopy using three diagnostic methods (direct smear in Lugol’s solution, decantation in distilled water and Fülleborn’s flotation). Results . Intestinal parasites were found in 144/386 of tested patients from the Ghazni province (37.3% infected, mainly with Ascaris lumbricoides , Giardia intestinalis , Hymenolepis nana ) and in 49/162 patients from the Parwan province (30.2% infected, mainly with G. intestinalis , A. lumbricoides , H. nana ). Conclusions . The rates of intestinal parasitic infections among Afghans are high. The wide range of the detected parasites (protozoa, nematodes, cestodes) should result in the introduction of general screening to be conducted regularly among inhabitants of Afghanistan and the application of targeted antiparasitic chemotherapy aiming to eliminate intestinal helminths and protozoa from the local community.

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.

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.

Introduction Alveolar echinococcosis is a parasitic disease caused by the larval stage of tapeworm Echinococcus multilocularis . It usually involves the liver, but can spread to other organs. The treatment of choice is a surgical resection supported by antiparasitic drugs. In the advanced stages of the disease a liver transplantation is the only option. Aim This article presents the problems related to care of patients after liver transplantation for advanced alveolar echinococcosis. Material Sixty-seven patients with alveolar echinococcosis were hospitalized in our clinic in the years 2000–2015. Liver transplantation has been a therapeutic option for 9 patients. We retrospectively analyzed data of qualification for the liver transplantation and the postoperative treatment. Results Follow-up time after liver transplantation ranged from 7 months to 155 months (average, 6.4 years). One patient, with a history of advanced disease (P4N1M0), died due to liver failure. One patient was lost to follow-up. After liver transplantation all patients were receiving albendazole treatment. Two patients did not follow the medical recommendations. In 1 patient, who decided to stop therapy after 1 year, the relapse of alveolar echinococcosis in the left lobe of the transplanted liver passing through the diaphragm to the pericardium was detected. In another case we suspected a relapse of alveolar echinococcosis in transplanted liver due to positive serological tests. Conclusion The prognosis of patient after liver transplantation for alveolar echinococcosis is good. The main problem caused by immunosuppressive therapy is a recurrence of disease in the transplanted liver.

Echinococcus multilocularis is a parasite that causes a dangerous zoonosis, alveolar echinococcosis (AE). Its presence in water sources, however, has scarcely been studied heretofore. Accordingly, 222 samples of different origin including wastewater from wastewater treatment plants (WWTPs) (n = 137), slaughterhouse (n = 49) as well as water from rivers (n = 26) and a cattle farm (n = 10) were collected from Xining City and a rural area in Qinghai-Tibet Plateau (QTP), an endemic area. Material obtained after processing of 10 L volume samples was subsequently analysed using three molecular detection methods: nested PCR, real-time PCR and LAMP. E. multilocularis DNA was found in 13 (5.85%) water samples; including 8 (5.8%), 3 (6%), 2 (20%) and 0 positive samples found in WWTPs, a slaughterhouse, a cattle farm and rivers, respectively. All three (LAMP, PCR, RT-PCR) molecular tools displayed high agreement and effectiveness in their ability of detecting the parasite’s DNA in environmental material. This is the first investigation describing E. multilocularis detection in wastewater samples, using three sensitive molecular diagnostic tools. Results indicate the role of wastewater in dissemination of E. multilocularis and the risk of contamination of water sources.

The CRISPR/Cas9 system, a natural defence system of bacterial organisms, has recently been used to modify genomes of the most important protozoa parasites. Successful genome manipulations with the CRISPR/Cas9 system are changing the present view of genetics in parasitology. The application of this system offers a major chance to overcome the current restriction in culturing, maintaining and analysing protozoan parasites, and allows dynamic analysis of parasite genes functions, leading to a better understanding of pathogenesis. CRISPR/Cas9 system will have a significant influence on the process of developing novel drugs and treatment strategies against protozoa parasites.

Malaria remains an important public health issue all over the world. Among 5 Plasmodium species invasive to humans, Plasmodium knowlesi has been identified most recently. It is sometimes difficult to differentiate this species from P. malariae with the use of microscopic examination. However, P. knowlesi infection may be associated with rapidly increasing parasitaemia and severe clinical course with the risk of death. Samples from Polish travellers returning from areas where simian malaria is endemic were examined with the use of polymerase chain reaction (PCR). The small subunit of ribosomal RNA (SSU rRNA) genes was subjected to analysis using nested PCR reaction. No positive results of P. knowlesi were obtained. Due to morphological similarities to P. malariae, potentially severe clinical course of infection and P. knowlesi endemic regions being a common tourist destination, diagnostic and clinical vigilance is necessary, including molecular methods use for precise parasite identification.

The aim of this study was to estimate the presence of Echinococcus multilocularis DNA in fruits, vegetables, and mushrooms in rural areas of Varmia-Masuria Province, Poland, which is the region with the highest number of human alveolar echinococcosis (AE) cases in this country. Recovery tests showed that E. multilocularis DNA is detectable in samples contaminated with at least 100 eggs of this tapeworm. In total, 103 environmental fruit, vegetable, and mushroom samples collected in forests, plantations, and kitchen gardens were analyzed using nested PCR assay based on the mitochondrial 12S ribosomal RNA (rRNA) gene. The parasite DNA was detected in 23.3 % of the samples. Sequencing confirmed that the obtained PCR products represented E. multilocularis. This study is the first environmental survey of the presence of E. multilocularis DNA in fruits, vegetables, and mushrooms intended for consumption. The results clearly demonstrate that it may be a direct source of human infections and shows the need to educate the public about the threat, especially people living in at-risk areas.

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.

This article presents the results of a preliminary study concerning cases of Chlamydia trachomatis infections among professional soldiers in the Polish Armed Forces. Soldiers who declared casual sexual contact with women were investigated in this study regarding the transmission of chlamydial infections by sexual activity. In total, 66 healthy, sexually active professional Polish soldiers, aged between 27 and 44, who didn't report any symptoms of urogenital infection were investigated. Urine samples taken from these patients were investigated using molecular methods (Cobas TaqMan, real-time PCR) in March-April 2012 in the Military Institute of Medicine, Warsaw, Poland. In the investigated group of 66 Polish soldiers, two 33-34 year old men were asymptomatic carriers of Chlamydia trachomatis (3.0%). They confirmed having casual sexual activity without prevention with many women. In the examined group of Polish soldiers the relatively low level of chlamydial infections found may result from the use of preventive measures during sexual activity. According to the examined men, the common use of condoms is mainly connected with the fear of HIV infection. Screening tests for Chlamydia trachomatis in the Polish Armed Forces are not performed, therefore incidence rates of chlamydial infections remain unknown. The authors plan further investigations with a larger group of professional soldiers.

The aim of this study was to estimate the presence of Echinococcus multilocularis DNA in the soil in rural areas of Varmia-Masuria Province, Poland, which is the region with the highest recorded number of human alveolar echinococcosis (AE) cases in this country. In total, 62 soil samples were analyzed. Samples were collected in forests near the lairs of foxes, as well as from arable fields, kitchen gardens and farmyards. The parasite DNA was detected in 11.3% of the samples. This study is the first environmental survey of the presence of E. multilocularis DNA in soil. The results demonstrate that the soil can be a direct source of human infections. These results should motivate the appropriate institutions to introduce informative and educational campaigns about the risk of infection, as well as developing methods for protecting the population, especially those living in at-risk areas.

Background: Members of the Polish Military Contingent (PMC) have been stationed in Afghanistan since 2002. They typically serve in areas characterised by low standards of sanitation which often leads to the development of food- and waterborne diseases. The aim of the study was to evaluate the prevalence of Giardia intestinalis infections among Polish soldiers deployed to Afghanistan. The research study was conducted as part of a programme for prevention of parasitic diseases of the gastrointestinal tract run by the Polish Armed Forces. Materials and methods: The study was carried out in August 2011; it involved 630 asymptomatic Polish soldiers serving in the Forward Operational Base (FOB) Ghazni in eastern Afghanistan. Stool specimens obtained from members of the PMC were first tested in FOB Ghazni (detection of Giardia intestinalis by Rida Quick Giardia immunochromatographic tests and Ridascreen Giardia immunoenzymatic tests — single samples). Next, the same biological material and two other faecal specimens fixed in 10% formalin were transported to the Military Institute of Medicine in Poland, where they were tested for Giardia intestinalis under light microscopy (direct smear, decantation in distilled water). Results: Parasitological tests performed under light microscopy showed that 2.7% (17/630) of the study group were infected with Giardia intestinalis. Some of these results were confirmed by immunochromatographic tests (6/630). In contrast, immunoenzymatic tests (ELISA) demonstrated a significantly higher detection rate reaching 18.1% (114/630). Immunoenzymatic tests confirmed all the positive results given by light microscopy and by immunochromatographic tests. Conclusions: The prevalence rate of Giardia intestinalis infections in Polish soldiers deployed to Afghanistan was found to be high. Microscopic methods exhibit low sensitivity and therefore may result in the underestimation of the true parasite prevalence. Immunoenzymatic tests (ELISA) showing a much higher sensitivity in comparison to light microscopy and immunochromatographic tests ought to be applied in screening for intestinal protozoan infections in areas characterised by harsh environmental conditions.

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.

Objective: To estimate the prevalence of intestinal parasites and their species in Afghan school children and to establish appropriate treatment methods for detected pathogens. Methods: Parasitological examination of stool samples collected from 1369 children aged 8-18, students of the Jahan Malika High School in Ghazni Province in eastern Afghanistan, was conducted in the period November 2013-April 2014. Three stool samples were collected from each patient every second day; the samples were fixed in 10% formalin and tested by light microscopy using the methods of direct smear in Lugol’s solution, decantation in distilled water, and Fülleborn’s flotation. Results: Of 535 examined children (39.1% of the study group) were infected with nematodes (n=324), cestodes (n=118), trematodes (n=12), and protozoa (n=228), 132 were diagnosed with co-infections (mainly ascariasis+giardiasis, ascariasis+hymenolepiasis) and received single or combined therapy. Conclusions: The Afghan community is an example of population characterized by a high rate of parasitic infections. Owing to high prevalence of multiple infections among inhabitants of Afghanistan, it seems that a mass deworming campaign with a single-dose chemotherapy may prove ineffective in eradicating intestinal parasites in the local population.

Toxoplasma gondii infections are prevalent in humans and animals all over the world. The aim of the study was to estimate the occurrence of T. gondii oocysts in fruits and vegetables and determine the genotype of the parasites. A total number of 216 fruits and vegetables samples were taken from shops and home gardens located in the area of northern Poland. Oocysts were recovered with the flocculation method. Then, real-time polymerase chain reaction (PCR) targeting the B1 gene was used for specific T. gondii detection and quantification. Toxoplasma DNA was found in 21 samples. Genotyping at the SAG2 locus showed SAG2 type I and SAG2 type II. This is the first investigation describing T. gondii DNA identification in a large number of fruits and vegetables samples with rapid molecular detection methods. The results showed that fruits and vegetables contaminated with T. gondii may play a role in the prevalence of toxoplasmosis in Poland.

Infections caused by Toxoplasma gondii are prevalent in humans and animals throughout the world. So far, there is no sufficient information concerning T. gondii oocysts prevalence in the environment, especially in soil. Therefore, the aim of this study was to estimate occurrence of T. gondii oocysts in soil and determine the genotype of detected parasites. A total of 101 soil samples were taken from different sites (sand-pits, "farming ground", areas around rubbish dumps) located in the Tri-City (Poland). Oocysts were recovered using the flotation method. Then, PCR reactions targeting the B1 gene were performed for specific T. gondii detection. The positive samples were further confirmed by PCR amplification of a repetitive element (REP) sequence [GenBank accession number AF146527]. Toxoplasma DNA was found in 18 samples. Among them, seven samples were successfully genotyped at the SAG2 locus. They were classified as SAG2 type I (5 samples) and SAG2 type II (2 samples). This is one of the first investigations describing T. gondii oocyst detection in environmental soil samples with rapid molecular detection methods and genotyping. The results of our findings showed that soil contaminated with T. gondii oocysts may play a role in the epidemiology of human toxoplasmosis in Poland.

Background The epidemiological situation concerning the prevalence of the majority of intestinal parasites in Poland has not been investigated in recent years and therefore is not known. Information on the prevalence of nematodes and most of cestodes is acquired from the studies carried out in Polish research centres. The aim of this study was to evaluate the occurrence of intestinal parasites among Polish soldiers who served in harsh environmental conditions in Afghanistan and their families living in Poland. The relation between the rates of infections in military personnel and in their families in the context of the risk of importing intestinal parasites from Afghanistan to Poland was also analysed in this article. Materials and methods 264 families consisting of 803 persons took part in the study which was conducted in the period May 2011 - December 2012. Stool samples were subjected to parasitological examination in the Military Institute of Medicine, Poland. The tests were performed by means of three diagnostic methods (direct smear, decantation, flotation). The study population comprised 264 married soldiers and 539 civilians (soldiers' wives and children) living permanently in urban areas in Poland. Results Intestinal parasites were diagnosed in 4/264 studied soldiers (1.5% infected with Ascaris lumbricoides, Giardia intestinalis), in 9/253 studied women (3.6% infected with A. lumbricoides, Hymenolepis nana, Taenia spp.) and in 5/286 children (1.7% infected with A. lumbricoides, G. intestinalis). There was no more than one infection per one household. Conclusions The occurrence of intestinal parasites among Polish soldiers shows the possibility of importing pathogens from the Third World countries. It also indicates, in the context of infected women and children, that the source of infection of parasites of the digestive tract can be present in Poland.

Respiratory tract infections (RTIs), beside diarrheas, skin lesions, and fevers of unknown origin, are one of the most common health problems acquired by travelers going to tropical and subtropical countries. Visitors to African, Asian, or South American destinations, typically characterized by harsh environmental conditions and poor sanitation standards, are at risk of exposure to a large number of pathogens causing infectious diseases. The infections are transmitted from contaminated food and water, through the air, direct contact, or by insects. The main modes of RTIs transmission include droplet infection and direct contact. The clinical spectrum of RTIs in travelers is broad, from upper respiratory tract infections, pharyngitis, bronchitis, pneumonia, to influenza-like illness. The spectrum of microbial agents causing respiratory infections include numerous viruses and bacteria, rarely fungi, and parasites. Most travelers complain of mild infections, only a small minority seek medical assistance and report to health care facilities. Because of the risk of importing pathogens into Europe or North America and transferring them onto the local population, it is important to present the scale of the problem in relation to rapid development of tourism industry and an increasing number of intercontinental journeys. The aim of the study was to discuss the occurrence of travel-related respiratory infections among representatives of temperate climate traveling to and returning from the tropics.

Highlights • Soldiers in combat zones are at risk of contracting respiratory infections. • Troops deployed to the harsh environment have high rates of respiratory symptoms. • Over 40% of soldiers’ respiratory illnesses have been due to unknown causative agents. • Limited diagnostic capabilities make it difficult to estimate the number of cases., Military personnel fighting in contemporary battlefields as well as those participating in combat training are at risk of contracting respiratory infections. Epidemiological studies have demonstrated that soldiers deployed to the harsh environment have higher rates of newly reported respiratory symptoms than non-deployers. Acute respiratory diseases are the principle reason for outpatient treatment and hospitalization among military personnel, with an incidence exceeding that of the adult civilian population by up to three-fold. Adenoviruses, influenza A and B viruses, Streptococcus pneumoniae, Streptococcus pyogenes, coronaviruses and rhinoviruses have been identified as the main causes of acute respiratory infections among the military population. Although infective pathogens have been extensively studied, a significant proportion of illnesses (over 40%) have been due to unknown causative agents. Other health hazards, which can lead to respiratory illnesses among troops, are extreme air temperatures, desert dust, emissions from burn pits, industrial pollutants, and airborne contaminants originating from degraded soil. Limited diagnostic capabilities, especially inside the area of operations, make it difficult to accurately estimate the exact number of respiratory diseases in the military environment. The aim of the study was to discuss the occurrence of respiratory tract infections in army personnel, existing risk factors and preventive measures.

Background This study concerns parasitological investigations estimating the prevalence of intestinal parasites in the Afghan community based on the example of patients, both children and adults, treated in the Ghazni Provincial Hospital, in the eastern part of the country. Materials and methods In total, 201 admitted patients with internal health problems were examined in the Afghan hospital in March 2012, including 164 children (1-17 years old) and 37 adults aged 18-80. Stool samples were tested in the Department of Epidemiology and Tropical Medicine at the Military Institute of Medicine in Gdynia, Poland using 3 parasitological methods: direct smear, decantation, flotation (light microscopy). Results Intestinal parasitic infections caused mainly by Ascaris lumbricoides, Giardia intestinalis and Hymenolepis nana were confirmed in 81/164 (49.4%) children examined. Among adults, 9/37 (24.3%) patients were infected with intestinal parasites including Ascaris lumbricoides and Giardia intestinalis. Conclusions The Afghan community, living in poor sanitary conditions with limited access to health services, is one of the most infected populations in the world. The parasitological investigation focused on intestinal parasites performed by the Polish health service among the eastern Afghan inhabitants is still one of the few screening studies in this country.

Minimisation of blood transmitted diseases is a basic element of all blood transfusion strategies. Civilian health service standards used in peacetime may be difficult to implement in a battlefield. The risk of blood -borne diseases depends on the applied donor qualification procedures and the epidemiological situation in the areas of military operations. The authors discuss various epidemiological aspects considered when selecting potential donors of fresh whole blood for a Walking Blood Bank at the Polish Field Hospital in Afghanistan.

Respiratory tract infections (RTIs), beside diarrheas, skin lesions, and fevers of unknown origin, are one of the most common health problems acquired by travelers going to tropical and subtropical countries. Visitors to African, Asian, or South American destinations, typically characterized by harsh environmental conditions and poor sanitation standards, are at risk of exposure to a large number of pathogens causing infectious diseases. The infections are transmitted from contaminated food and water, through the air, direct contact, or by insects. The main modes of RTIs transmission include droplet infection and direct contact. The clinical spectrum of RTIs in travelers is broad, from upper respiratory tract infections, pharyngitis, bronchitis, pneumonia, to influenza-like illness. The spectrum of microbial agents causing respiratory infections include numerous viruses and bacteria, rarely fungi, and parasites. Most travelers complain of mild infections, only a small minority seek medical assistance and report to health care facilities. Because of the risk of importing pathogens into Europe or North America and transferring them onto the local population, it is important to present the scale of the problem in relation to rapid development of tourism industry and an increasing number of intercontinental journeys. The aim of the study was to discuss the occurrence of travel-related respiratory infections among representatives of temperate climate traveling to and returning from the tropics.

Leishmania parasites are the etiological agents of leishmaniosis, with severe course and often fatal prognosis, and the global number of cases has increased in recent decades. The gold standards for the diagnosis of leishmaniosis are microscopic examinations and culture in vitro of the different clinical specimens. The sensitivity of these methods is insufficient. Recent development in specific and sensitive molecular methods (PCR) allows for detection as well as identification of the parasite species (subspecies). The aim of the study was to estimate the usefulness of molecular methods (PCR) for detection of Leishmania species and consequently for the implementation of such methods in routine diagnostics of leishmaniosis in Polish patients returning from endemic areas of the disease. In our investigations we used 54 known Leishmania positive DNA templates (from culture and clinical specimens) received from the CDC (Atlanta, GA, USA). Moreover, 25 samples of bone marrow, blood or other tissues obtained from 18 Polish individuals suspected of leishmaniosis were also examined. In PCR we used two pairs of primers specific to the conserved region of Leishmania kinetoplast DNA (kDNA) minicircle (13A/13B and F/R). Using these primers we obtained amplicons in all DNA templates from the CDC and in three Polish patients suspected for Leishmania infection. In one sample from among these cases we also obtained positive results with DNA isolated from a blood specimen which was previously negative in microscopic examinations.

Purpose: To estimate the prevalence and species of intestinal parasites in an Afghan community as well as to establish appropriate treatment method for each of the detected pathogens. Methods: Parasitological examination of stool samples collected from 777 children aged 1-17 years, treated in Ghazni Provincial Hospital (eastern Afghanistan) conducted in the period 2012 - 2013 made it possible to implement dedicated antiparasitic treatment. Results: Of 312 children (40.2 % of the examined group; p < 0.05) infected with nematodes (n = 154, 19.7 %), cestodes (n = 90, 11.6 %), trematodes (n = 16, 2.1 %), and protozoa (n = 135, 17.4 %), 67 were diagnosed with co-infections (mainly ascariasis + giardiasis, ascariasis + hymenolepiasis, giardiasis + hymenolepiasis, ascariasis + giardiasis + hymenolepiasis) and received complex therapy (albendazole + metronidazole, albendazole + praziquantel, metronidazole + praziquantel, and albendazole + metronidazole + praziquantel). Conclusion: Owing to high prevalence of multiple co-infections among inhabitants of the Afghan community, it seems that a mass deworming campaign with a single-dose chemotherapy (albendazole 400 mg or mebendazole 500 mg) may prove ineffective in eradicating intestinal parasites in the local population. Keywords: Afghanistan, Intestinal parasites, Hymenolepiasis, Ascariasis, Giardiasis, Metronidazole, Albendazole, Praziquantel, Mebendazole, Deworming

Anaplasma phagocytophilum and Babesia spp. are emerging tick-borne pathogens which can threaten human health. A duplex real-time PCR and qPCRs with primers and probes targeting 97 and 116 bp fragments of 16S rRNA and 18S rRNA genes, respectively, were used for qualitative and quantitative detection of both pathogens in Ixodes ricinus ticks. Altogether 1875 ticks (1084 adults and 791 nymphs) were collected from rural and urban habitats in northern Poland. Of them, at least 0.9% were found to be infected with A. phagocytophilum while 2.5% with Babesia spp. A comparison of the infection rates by the tick stage, the type of area, the collection site, habitats of different tick density and by the month of collection was done. The prevalence of pathogens was significantly lower in nymphs than in adult ticks (p = 0.02) and in rural areas than in urban areas (p = 0.007). Four different 16S rRNA gene variants of A. phagocytophilum were determine, however none of them showed 100% identity with compared sequences isolated from human patients. The dominant Babesia species was B. venatorum. Results of qPCRs with circular and linearized forms of plasmids used as the standards showed significant difference in the pathogen loads (p = 0.001). The copy numbers of A. phagocytophilum and Babesia spp. estimated from the linear plasmids were 28.7 and 5.1 times lower, respectively, when compared with their circular forms, and were accepted as more reliable. The average number of copies of 16S rRNA gene of A. phagocytophilum in the positive I. ricinus samples were 3.39 × 10(5) ± 6.09 × 10(5). The mean copy number of 18S rRNA gene of Babesia spp. was ~2.55 × 10(5) ± 1.04 × 10(6). We confirmed the presence of A. phagocytophilum and Babesia spp. in I. ricinus in both rural and urban environments. The determined low infection rates suggests, however, that the risk for local population and tourists to acquire infection is also low. Moreover, we confirmed recent findings that serious overestimation by circular plasmid DNA makes it less suitable as a standard and that the linear standards should be recommended for qPCR.

Different species of amoebae belonging to the genus Acanthamoeba are widely distributed in many parts of the world and known as free-living organisms. Some strains of the protozoans may exist as parasites and cause risk to human health as causative agents of serious human diseases. Currently, in Poland, there is no sufficient information about the distribution of Acanthamoeba strains and their genotypes in the environment. Therefore, 20 environmental surface water samples were collected from different sites located at five water reservoirs in Gdynia, Sopot, and Gdansk (northern Poland). The material was cultured to obtain Acanthamoeba isolates that were then specifically analyzed with both PCR and real-time PCR assays. Of the 20 samples examined, Acanthamoeba DNA was found in 13 samples tested with the use of real-time PCR; in 10 of them, DNA of the amoeba was also detected using PCR technique. The comparison with sequences available in the GenBank confirmed that the PCR products are fragments of Acanthamoeba 18S rRNA gene and that isolates represent T4 genotype, known as the most common strains related to AK cases. This is the first investigation in Poland describing Acanthamoeba detection in environmental water samples with molecular techniques and genotyping. The results indicate that surface water in Poland may be a source of acanthamoebic strains potentially pathogenic for humans.