Protozoa Of Genus Research Articles

Assessment of the effect of silver nanoparticles on the relevant soil protozoan genus Acanthamoeba

AbstractSilver nanoparticles (AgNP) are used in a broad range of consumer products and industrial applications. During the regular product life cycle and disposal, AgNP are continuously released into the environment. Hence, the aim of this study was to investigate the potential ecotoxicological effects of AgNP exposure on amoebae. The Acanthamoeba castellanii ATCC 30234 strain and environmental isolate Acanthamoeba strain C5/2, which are both affiliated with genotype T4, were chosen as representatives of ecologically important soil protozoan organisms. The amoebae were exposed to citrate‐stabilized AgNP (30 and 70 nm in size) for 24 h and 96 h at concentrations ranging from 600 µg L−1 to 20 mg L−1. A newly adopted cell culture based microscopic assay was applied to assess the adherence ability of the amoeba trophozoites. The general metabolic activity of Acanthamoeba was determined to be a second independent endpoint by means of intracellular reduction of the redox dye AlamarBlue®. The fate of AgNP within the amoebae and test solutions was visualized by light‐ and transmission electron microscopy (TEM). Both Acanthamoeba strains showed a significant dose‐dependent decrease of adherence ability (p < 0.04) and metabolic activity (p < 0.01) after 96 h of AgNP exposure. The environmental Acanthamoeba strain C5/2 lost both its adherence ability and metabolic activity at lower AgNP concentrations than the type strain, indicating a higher sensitivity to ionic silver. This was confirmed by the application of AgNO3, provoking a higher effect level in strain C5/2. AgNP was visualized intracellularly by transmission electron microscopy within the cytoplasm of Acanthamoeba. This is the first report to show the ecotoxicological effects of short‐term AgNP exposure on the soil protist Acanthamoeba, causing both changes in the adherence ability and metabolic activity of this amoeba. This combined approach may be a powerful tool in the future for predicting potential harmful ecotoxicological effects of AgNP exposure using soil protozoans.

Індикація та видова диференціація найпростіших роду Babesia за методом ПЛР у кліщах, знятих з тварин

The article presents the study results on the indication and species differentiation of the Babesia genus protozoa in the organism of ticks taken off dogs and cattle. Diagnostic tests were performed using a multiplex PCR test system, being a self-engineering product, which allows to determine the DNA presence of 6 Babesia genus species in any biological samples, namely: B. canis, B. divergens, B. caballi, B. major, B Bigemina, B. bovis, three of them being species-specific, by the bands’ size at the amplification products’ electrophoregrams. The above test system contains 2 direct and 3 reverse primers flanking the DNA fragments of the gene encoding the 18S rRNA of the Babesia genus protozoa.The subject of the study were ticks obtained from 17 dogs of different breeds and sex-age status and 12 units of livestock (cattle). As a result, the babesial DNA was detected in 11 samples, eight of which were differentiated as Babesia canis, two as Babesia bovis and one as Babesia divergens. Based on the results obtained, the animals, whose ticks, taken off them, had babesia detected, one or another strategy was developed for treatment and prophylaxis measures.The approach suggested in the present article permits identifying the possible risks of babesia invasion at the earliest stages of the disease. Presence or absence of the Babesia genus protozoa in the body of sanguivorous ticks permits avoiding treatments rather toxic for the organism, or to carry out preventive therapy long before the babesiosis clinical symptoms’ manifestation. It is vitally important for pregnant females and especially sensitive to babesiosis dog breeds.The prospect of further research is development of the PCR test systems for indicating and differentiating other tick-borne infections and invasions.

An investigation of parasitic infections and review of molecular characterization of the intestinal protozoa in nonhuman primates in China from 2009 to 2015.

Parasites are a well-known threat to nonhuman primate (NHP) populations, and potentially cause zoonotic diseases in humans. In this study, the basic data was provided of the parasites in NHPs and the molecular characterization of the Enterocytozoon bieneusi, Giardia duodenalis, Cryptosporidium spp., and Entamoeba spp. were reviewed, which were found in these samples. A total of 3349 fecal samples were collected from 34 species reared at 17 districts in zoos, farms, free-range, or research laboratories, and examined microscopically. Eleven genera of intestinal parasites were detected: five genera of protozoans (Isospora spp., Entamoeba spp., Giardia sp., Cryptosporidium spp., and Cyclospora spp.) and six genera of helminths (Trichuris spp., Strongyloides spp., Ascaris spp., Physaloptera spp., Ancylostoma spp., and Enterobius spp.). The overall sample prevalence of parasitic infection was 54.1% (1811/3349). Entamoeba spp. was the most prevalent (36.4%, 1218/3349). The infection rate was the highest in free-range animals (73.0%, 670/918) (P < 0.01) and Guangxi Zhuang autonomous region (64.8%, 566/873). Mixed infections were mostly detected for Entamoeba spp., Trichuris spp., and Strongyloides spp.. Molecular characterization was reviewed of Enterocytozoon bieneusi, Giardia duodenalis, Cryptosporidium spp., and Entamoeba spp., as these are zoonotic species or genotypes. This parasitological data for NHPs in China, provides important information for veterinarians and public health authorities for the elimination of such parasites and monitor the potential transmission of zoonotic infections from NHPs.

Differences in immune responses against Leishmania induced by infection and by immunization with killed parasite antigen: implications for vaccine discovery.

The leishmaniases are a group of diseases caused by different species of the protozoan genus Leishmania and transmitted by sand fly vectors. They are a major public health problem in almost all continents. There is no effective control of leishmaniasis and its geographical distribution is expanding in many countries. Great effort has been made by many scientists to develop a vaccine against leishmaniasis, but, so far, there is still no effective vaccine against the disease. The only way to generate protective immunity against leishmaniasis in humans is leishmanization, consisting of the inoculation of live virulent Leishmania as a means to acquire long-lasting immunity against subsequent infections. At present, all that we know about human immune responses to Leishmania induced by immunization with killed parasite antigens came from studies with first generation candidate vaccines (killed promastigote extracts). In the few occasions that the T cell-mediated immune responses to Leishmania induced by infection and immunization with killed parasite antigens were compared, important differences were found both in humans and in animals. This review discusses these differences and their relevance to the development of a vaccine against leishmaniasis, the major problems involved in this task, the recent prospects for the selection of candidate antigens and the use of attenuated Leishmania as live vaccines.

Prevalence of intestinal parasites in breeding cattery cats in Japan.

To address the lack of up-to-date published data, the present study assessed the prevalence of intestinal parasites in breeding catteries in Japan. Fresh faecal samples were randomly collected from 342 cats (aged 1 month to 12 years) in seven breeding catteries in Japan, located in prefectures of Nagano (n = 2), Saitama (n = 1), Aichi (n = 2), Gifu (n = 1) and Miyagi (n = 1), on a single occasion. The samples were tested for the presence of Giardia species copro-antigen using a commercially available enzyme-linked immunosorbent assay kit. Other intestinal parasites were identified microscopically using the formalin-ethyl acetate sedimentation technique. The total prevalence of intestinal parasites was 20.8%; only two genera of protozoa (Giardia species: 18.7% and Cystoisospora species: 5.0%) were detected. Coinfections of both protozoans were recorded in 2.9% of cats. In contrast, no helminths were detected. The presence of total infection, Giardia species, Cystoisospora species and multiple infections in cats <1 year old were significantly more prevalent than in cats ⩾1 year old. There were no significant differences among faecal conditions with or without intestinal parasites. Giardia species infection was present in samples from all breeding catteries, except for one facility. Cystoisospora species and coinfections were shown in four and two breeding catteries, respectively. The prevalence of intestinal parasites was markedly variable among the breeding catteries. The present study demonstrates the significance of Giardia species and Cystoisospora species infections in breeding cattery cats. Additionally, it is suggested that environmental contamination is the most important factor influencing the prevalence of protozoal infections in breeding catteries.

A review of canine babesiosis: the European perspective.

Canine babesiosis is a significant tick-borne disease caused by various species of the protozoan genus Babesia. Although it occurs worldwide, data relating to European infections have now been collected for many years. These data have boosted the publication record and increased our working knowledge of these protozoan parasites. Both the large and small forms of Babesia species (B. canis, B. vogeli, B. gibsoni, and B. microti-like isolates also referred to as "B. vulpes" and "Theileria annae") infect dogs in Europe, and their geographical distribution, transmission, clinical signs, treatment, and prognosis vary widely for each species. The goal of this review is to provide veterinary practitioners with practical guidelines for the diagnosis, treatment and prevention of babesiosis in European dogs. Our hope is that these guidelines will answer the most frequently asked questions posed by veterinary practitioners.

Dispensability of the [4Fe‐4S] cluster in novel homologues of adenine glycosylase MutY

7,8-Dihydro-8-deoxyguanine (8oG) is one of the most common oxidative lesions in DNA. DNA polymerases misincorporate an adenine across from this lesion. Thus, 8oG is a highly mutagenic lesion responsible for G:C→T:A transversions. MutY is an adenine glycosylase, part of the base excision repair pathway that removes adenines, when mispaired with 8oG or guanine. Its catalytic domain includes a [4Fe-4S] cluster motif coordinated by cysteinyl ligands. When this cluster is absent, MutY activity is depleted and several studies concluded that the [4Fe-4S] cluster motif is an indispensable component for DNA binding, substrate recognition and enzymatic activity. In the present study, we identified 46 MutY homologues that lack the canonical cysteinyl ligands, suggesting an absence of the [4Fe-4S] cluster. A phylogenetic analysis groups these novel MutYs into two different clades. One clade is exclusive of the order Lactobacillales and another clade has a mixed composition of anaerobic and microaerophilic bacteria and species from the protozoan genus Entamoeba. Structural modeling and sequence analysis suggests that the loss of the [4Fe-4S] cluster is compensated by a convergent solution in which bulky amino acids substitute the [4Fe-4S] cluster. We functionally characterized MutYs from Lactobacillus brevis and Entamoeba histolytica as representative members from each clade and found that both enzymes are active adenine glycosylases. Furthermore, chimeric glycosylases, in which the [4Fe-4S] cluster of Escherichia coli MutY is replaced by the corresponding amino acids of LbY and EhY, are also active. Our data indicates that the [4Fe-4S] cluster plays a structural role in MutYs and evidences the existence of alternative functional solutions in nature.

The diversity and expansion of the trans-sialidase gene family is a common feature in Trypanosoma cruzi clade members

Trans-sialidase (TS) is a polymorphic protein superfamily described in members of the protozoan genus Trypanosoma. Of the eight TS groups recently described, TS group I proteins (some of which have catalytic activity) are present in the distantly related Trypanosoma brucei and Trypanosoma cruzi phylogenetic clades, whereas other TS groups have only been described in some species belonging to the T. cruzi clade. In the present study we analyzed the repertoire, distribution and phylogenetic relationships of TS genes among species of the T. cruzi clade based on sequence similarity, multiple sequence alignment and tree-reconstruction approaches using TS sequences obtained with the aid of PCR-based strategies or retrieved from genome databases. We included the following representative isolates of the T. cruzi clade from South America: T. cruzi, T. cruzi Tcbat, Trypanosoma cruzi marinkellei, Trypanosoma dionisii, Trypanosoma rangeli and Trypanosoma conorhini. The cloned sequences encoded conserved TS protein motifs Asp-box and VTVxNVxLYNR but lacked the FRIP motif (conserved in TS group I). The T. conorhini sequences were the most divergent. The hybridization patterns of TS probes with chromosomal bands confirmed the abundance of these sequences in species in the T. cruzi clade. Divergence and relationship analysis placed most of the TS sequences in the groups defined in T. cruzi. Further examination of members of TS group II, which includes T. cruzi surface glycoproteins implicated in host cell attachment and invasion, showed that sequences of T. cruzi Tcbat grouped with those of T. cruzi genotype TcI. Our analysis indicates that different members of the T. cruzi clade, with different vertebrate hosts, vectors and pathogenicity, share the extensive expansion and sequence diversification of the TS gene family. Altogether, our results are congruent with the evolutionary history of the T. cruzi clade and represent a contribution to the understanding of the molecular evolution and role of TS proteins in trypanosomes.

Molecular genotyping and sub-genotyping of Cryptosporidium spp. isolates from symptomatic individuals attending two major public hospitals in Madrid, Spain

Infections by members of the protozoan genus Cryptosporidium are among the most common causes of human gastrointestinal illness worldwide. In Spain cryptosporidiosis is not a compulsory notifiable disease, so the actual burden of the infection in both clinical and general populations remains largely unknown. We present here data on the diversity and frequency of the Cryptosporidium species and sub-genotypes identified in symptomatic individuals seeking medical care in two major hospitals in Madrid, Spain, between December 2013 and January 2015. Initial detection of the parasite was conducted on a total of 122 stool samples collected from 120 patients by microscopy with modified Ziehl-Neelsen and/or immunochromatographic tests. We used immunofluorescence, PCR-based methods and sequence analyses of the 60-kDa (GP60) glycoprotein and the small subunit ribosomal RNA (SSU rRNA) genes for confirmatory purposes and to characterize Cryptosporidium isolates. A total of 110 patients were confirmed with cryptosporidiosis. Overall, 101 isolates were successfully sub-genotyped at the GP60 locus, and an additional seven at the SSU rRNA locus. The analyses of all amplicons defined 10 distinct sequence types representing the GP60 family sub-genotypes IbA10G2 (78.7%), IeA11G3T3 (3.7%) of C. hominis, and the GP60 family sub-types IIaA15G2R1 (5.6%), IIaA18G6R1 (0.9%), IIcA5G3a (0.9%), IIdA18G1 (0.9%), IIdA19G1 (0.9%), IIdA21G1 (0.9%), and IIdA22G1 (0.9%) of C. parvum. A single isolate was assigned to C. felis (0.9%), two C. parvum isolates (1.9%) could not be characterized at the sub-genotype level and an additional four isolates (3.7%) were not typable. These results strongly suggest that transmission of cryptosporidiosis is mostly anthroponotic in origin in the clinical sample under study. We expect that our molecular epidemiological data will make a significant contribution to unravel the actual epidemiological situation of cryptosporidiosis in Spain, providing health care and policy makers with solid baseline information to unavoidably improve the national surveillance system and allocate additional resources to research, diagnosis, and treatment of cryptosporidiosis.

The African buffalo parasite Theileria. sp. (buffalo) can infect and immortalize cattle leukocytes and encodes divergent orthologues ofTheileria parva antigen genes.

African Cape buffalo (Syncerus caffer) is the wildlife reservoir of multiple species within the apicomplexan protozoan genus Theileria, including Theileria parva which causes East coast fever in cattle. A parasite, which has not yet been formally named, known as Theileria sp. (buffalo) has been recognized as a potentially distinct species based on rDNA sequence, since 1993. We demonstrate using reverse line blot (RLB) and sequencing of 18S rDNA genes, that in an area where buffalo and cattle co-graze and there is a heavy tick challenge, T. sp. (buffalo) can frequently be isolated in culture from cattle leukocytes. We also show that T. sp. (buffalo), which is genetically very closely related to T. parva, according to 18s rDNA sequence, has a conserved orthologue of the polymorphic immunodominant molecule (PIM) that forms the basis of the diagnostic ELISA used for T. parva serological detection. Closely related orthologues of several CD8 T cell target antigen genes are also shared with T. parva. By contrast, orthologues of the T. parva p104 and the p67 sporozoite surface antigens could not be amplified by PCR from T. sp. (buffalo), using conserved primers designed from the corresponding T. parva sequences. Collectively the data re-emphasise doubts regarding the value of rDNA sequence data alone for defining apicomplexan species in the absence of additional data. ‘Deep 454 pyrosequencing’ of DNA from two Theileria sporozoite stabilates prepared from Rhipicephalus appendiculatus ticks fed on buffalo failed to detect T. sp. (buffalo). This strongly suggests that R. appendiculatus may not be a vector for T. sp. (buffalo). Collectively, the data provides further evidence that T. sp. (buffalo). is a distinct species from T. parva.

Molecular and chemical dialogues in bacteria-protozoa interactions.

Protozoan predation of bacteria can significantly affect soil microbial community composition and ecosystem functioning. Bacteria possess diverse defense strategies to resist or evade protozoan predation. For soil-dwelling Pseudomonas species, several secondary metabolites were proposed to provide protection against different protozoan genera. By combining whole-genome transcriptome analyses with (live) imaging mass spectrometry (IMS), we observed multiple changes in the molecular and chemical dialogues between Pseudomonas fluorescens and the protist Naegleria americana. Lipopeptide (LP) biosynthesis was induced in Pseudomonas upon protozoan grazing and LP accumulation transitioned from homogeneous distributions across bacterial colonies to site-specific accumulation at the bacteria-protist interface. Also putrescine biosynthesis was upregulated in P. fluorescens upon predation. We demonstrated that putrescine induces protozoan trophozoite encystment and adversely affects cyst viability. This multifaceted study provides new insights in common and strain-specific responses in bacteria-protozoa interactions, including responses that contribute to bacterial survival in highly competitive soil and rhizosphere environments.

A phylogeny of sand flies (Diptera: Psychodidae: Phlebotominae), using recent Ethiopian collections and a broad selection of publicly available DNA sequence data

AbstractSand flies in the psychodid subfamily Phlebotominae carry important human pathogens in the trypanosomatid protozoan genus Leishmania (Cupolillo). Despite the fact that hundreds of sequences for this group are now publicly available, they constitute different sets of taxa and genetic markers. Integrating these data to construct a molecular phylogeny of the family is a significant bioinformatics challenge. We used sequences of eight markers obtained from freshly collected sand flies from Ethiopia and combined them with over 1300 publicly available sequences, performing a combined analysis after generating single terminal sequences from ancestral reconstructions for some individual markers. The resulting phylogeny had 113 terminals and recovered Phlebotominae and certain species groups as monopheletic. Although the 20 outgroups in Psychodinae were recovered as a well‐resolved clade with bootstrap support for many internal clades, Phlebotominae was recovered as several lineages with unclear relationships among them. However, phlebotomines clustered by geographic region, the most notable being all the New World species except Brumptomyia (Galati), which were recovered as monophyletic. Our phylogeny suggests a Sub‐Saharan African or South Asian origin for the subfamily, which subsequently expanded to the north and west, and eventually to the New World. Supported species groups are often composed of widespread species with overlapping ranges. This result highlights the need for a large increase in the amount and diversity of molecular sequence data, and a broad selection of terminals, to test taxonomic hypotheses and examine speciation processes in this important group of flies.

¿Cumplir con la legislación nos garantiza consumir agua segura?

Current legislation on microbiological control of water for human consumption in our country, and in many others, does not include the search of human-pathogenic parasites or viruses. Even though water purification methods used for the removal of bacteria from water do also remove the great majority of parasitic forms, some of their resistance elements are not affected by chlorination treatments. Groundwater obtained from shallow wells in semi-rural areas may also constitute a risk since they are very susceptible to contamination.The aim of this work was to quantify parasitic elements and viruses present in water for human consumption in the Province of Salta, in Argentina. For that, 60 L water samples were collected in six points located in the Province, including two sources of water: the distribution network and shallow wells. Samples were collected during dry and wet seasons. Ultrafiltration techniques were used to concentrate the water in pathogens. Optical microscopy using a Neubauer chamber for the quantification of parasite elements and real-time PCR for the quantitative detection of viruses were used.We found a great variety of protozoa genera in both groundwater and in the distribution network. Only in groundwater samples we found helminths and in one of them also adenovirus. Results showed that there is little relation between the presence of bacterial indicators (established by legislation) and pathogens as parasites and viruses. Numerous water samples that are within the limits established by legislation (for which they would be considered acceptable as water for human consumption) had pathogenic parasites or viruses. This shows the urgent need to update the current legislation to include parasitic indicators that allow certifying that the water is safe for human consumption, therefore decreasing the risk of waterborne diseases.

Abundance of zooplankton and physico-chemical parameters of a polyculture fish pond of Manikganj, Bangladesh

Composition and abundance of zooplankton in a polyculture pond was studied during June 2003 to May 2004. A total of 33 major species of zooplankton including 11genera of Rotifera, 4 of Copepoda, 4 genera of Cladocera and 2 genera of Protozoa were identified. The monthly abundance of zooplankton varied form 4228 indiv/l in October 2003 to169 indiv/l in January 2004. Among the total zooplankton, Rotifera comprised of 71.07%, Copepoda comprised of 12.49% Nauplii comprised of 11.96%, Cladocera comprised of 1.49% and Protozoa comprised of 2.99%. Water temperature (r = 0.18), free CO2 (r = 0.40), TDS (r = 0.14) and water depth (r = 0.55) showed positive correlation with zooplankton but air temperature, pH, dissolved O2, total hardness and ammonia showed negative correlation (the coefficients being -0.18, -0.05, -0.25, -0.38 and -0.11 respectively) with the occurrence of zooplankton. The average growth of fishes and zooplankton showed positive correlation in all cases but the relations were insignificant when tested statistically.Bangladesh J. Zool. 42(1): 67-76, 2014

Diversity of Cryptosporidium in brush-tailed rock-wallabies (Petrogale penicillata) managed within a species recovery programme.

Host-parasite relationships are likely to be impacted by conservation management practices, potentially increasing the susceptibility of wildlife to emerging disease. Cryptosporidium, a parasitic protozoan genus comprising host-adapted and host-specific species, was used as an indicator of parasite movement between populations of a threatened marsupial, the brush-tailed rock-wallaby (Petrogale penicillata). PCR screening of faecal samples (n = 324) from seven wallaby populations across New South Wales, identified Cryptosporidium in 7.1% of samples. The sampled populations were characterised as captive, supplemented and wild populations. No significant difference was found in Cryptosporidium detection between each of the three population categories. The positive samples, detected using 18S rRNA screening, were amplified using the actin and gp60 loci. Multi-locus sequence analysis revealed the presence of Cryptosporidium fayeri, a marsupial-specific species, and C. meleagridis, which has a broad host range, in samples from the three population categories. Cryptosporidium meleagridis has not been previously reported in marsupials and hence the pathogenicity of this species to brush-tailed rock-wallabies is unknown. Based on these findings, we recommend further study into Cryptosporidium in animals undergoing conservation management, as well as surveying wild animals in release areas, to further understand the diversity and epidemiology of this parasite in threatened wildlife.

Preliminary report on the intestinal parasites and their diversity in captive Chinese alligators.

Although intestinal parasites are commonly detected in either wild or farmed crocodiles in other parts of the world, yet few reports have ever been on the captive bred Chinese alligators (Alligator sinensis) since they are only distributed in the southern areas of the Yangtze River. The current work was undertaken to investigate the intestinal parasites and their diversity in the Chinese crocodilian species. In May and October of 2013, we randomly collected a total of 328 fecal samples of the captive Chinese alligators in a breeding centre in south Anhui province. Three genera of protozoa (Entaoeba, Eimeria and Isospora), five genera of nematodes (Ascaris, Dujardinascaris, Capillaria, Toxocara and Strongylus), two genera of trematodes (Echinostoma and Clonorchis) and two families of trematodes (Schistosomatidae and Cryptogonimidae) were identified, in which Dujardinascaris was the most prevalent and led to the highest infection rate (14.33%), and Entaoeba ranked the second (13.11%). The number of species, richness index, diversity index and evenness index were higher in July and August, and those indexes were relatively most in juvenile and subadult alligators. Our findings suggest that the intestinal parasites infection was prevalent in captive Chinese alligators, and this condition requires our attention, whatever it is for controlling or preventing the intestinal parasitic disease from spreading to humans or conservation of this endangered species.

Population analyses of microfauna at a Water treatment Plant at Shebeen Alkoom, Minufeya, Egypt.

Safe Drinking water is a priority for human health all over the world. Accordingly, the microfauna of raw, filtered, treated, and tap water were investigated in this study. The results proved the efficiency of the Water Plant, but slight occurrence of non-pathogenic bacteria in tap water, indicated the necessity to renovate the distribution system. Most of the physico-chemical characters are within the standards for drinking water apart from phosphate, nitrate, iron and manganese. Raw water samples only were positive for E. coli, T. & F. coliform, Salmonella and Shigella. This could be referred to the illegal discharge of sewage and other organic pollutants into the water source. Sixty one protozoan genera were identified only in raw water which can be attributed to the influence of chlorination during water treatment. Ciliated protozoan densities proved seasonal variation with highest values during Autumn. Protozoan monthly densities showed significant correlations with certain physical and chemical parameters. Therefore ciliates are considered as bio-indicators for pollution in various aquatic habitats. Simultaneously, rotifers can be used to indicate the performance level of certain stages of water treatment process as they showed significant correlations with different ecological factors.

Next generation sequencing as a novel tool for diagnostics of apicomplexan pathogen in ticks and mammalian hosts

Among apicomplexan parasites, ticks are known vector of several species belongs to three protozoan genera (Babesia, Theileria and Hepatozoon). During their life cycle, tick-transmitted apicomplexan parasites alternate between asexual (in vertebrate host) and sexual (in ticks) developmental stages. The major constraint for the proper diagnostics of those pathogens is the high possibility of mix infection, both in ticks and vertebrate hosts, with several species or genotypes. The aim of this study was to apply the Next Generation Sequencing (NGS) as a method of choice for simultaneous determination of the full spectrum of apicomplexan pathogens in ticks and the mammalian hosts. Therefore, A pair of universal primers were designed to flank a 167 bp barcode region of the 18s rRNA gene of all Babesia, Theileria and Hepatozoon species. The new protocol was evaluated on DNA samples isolated from 195 dogs and 144 ticks (Rhipicephalus armatus and R. pulchellus) collected from Northern Kenya. In total 301 sample (89%) were positive for apicomplexan infections; ranging from single to multiple infection with one species or several species and/or genotypes in a single sample. The most abundant apicomplexan pathogens were Hepatozoon followed by Babesia and Theileria, respectively. Further, the result shows that the barcode region entails enough variability that allows identifying the pathogens up to the subspecies and genotypes level. The exact methodological and results detailed will be presented later. This work was supported by the project OP VK CZ.1.07/2.3.00/30.0014.

Prevalence of intestinal parasites in pet shop kittens in Japan.

The present study examined the prevalence of intestinal parasites in kittens from five pet shops in East Japan. Fresh faecal samples were collected from 555 kittens (aged 1-3 months) on a single occasion. The samples were tested for the presence of Giardia species coproantigen using a commercially available enzyme-linked immunosorbent assay kit. Other intestinal parasites were identified microscopically using the formalin-ethyl acetate sedimentation technique. The overall prevalence of intestinal parasites was 27.2%; two genera of protozoa (Giardia species and Cystoisospora species) and one nematode (Toxocara cati) were detected. Faecal condition was not related to intestinal parasite infections. Significant differences among the pet shops were observed in the overall prevalence of intestinal parasites and the presence of Cystoisospora species infections.

Caracterización microbiológica y fisicoquímica de aguas subterráneas de los municipios de La Paz y San Diego, Cesar, Colombia

Dentro de las funciones de evaluación, control y seguimiento ambiental de los usos de las fuentes de agua existentes en el Cesar por parte de la Corporación Autónoma Regional del Cesar- CORPOCESAR, se realizó un estudio para evaluar la calidad de aguas subterráneas ubicadas en 93 aljibes en los municipios de La Paz y San Diego (Cesar) en 2009 de acuerdo con lo estipulado en el decreto 1575/07 y la resolución 2115/07 del Ministerio de Protección Social y el Ministerio de Ambiente, Vivienda y Desarrollo Territorial. Se caracterizaron propiedades microbiológicas (determinación de Pseudomona aeruginosa y protozoos patógenos) y fisicoquímicas (conductividad, pH, temperatura, sólidos disueltos totales, salinidad, acidez, alcalinidad, turbidez, cloruros, amonio, nitritos, nitratos, hierro, magnesio, sodio y calcio, estos tres últimos análisis necesarios para calcular la aptitud para riego con el fin de evidenciar la calidad del agua de la que se abastecen los habitantes de estos predios). La detección de P. aeruginosa se realizó mediante filtración por membrana y protozoos mediante la técnica de flotación por centrifugación con sulfato de zinc. De las muestras analizadas en un 84,94% se encontró P. aeruginosa. Se identificaron 5 géneros de protozoos siendo Giardia sp el patógeno con mayor prevalencia, representando un 46,1%, seguido de Criptosporidium sp con un 22,18%; en relación con los resultados fisicoquímicos, estos indicaron que de los aljibes muestreados solo un 4,3% contienen agua apta para riego sin que su uso acarree riesgos para la salud.