Human cryptosporidiosis in Iran: a systematic review and meta-analysis

Cryptosporidiosis caused by Cryptosporidium spp. is an important parasitic disease that can be life-threatening for children and immunocompromised patients. This systematic review and meta-analysis was designed to determine the prevalence rate of Cryptosporidium infection and related risk factors among the Iranian general population. We searched electronic databases including Google Scholar, PubMed, Science Direct, Scopus and Proquest for articles in English and SID, Magiran, IranMedex, and IranDoc for articles in Persian. Out of 4816 studies identified in the electronic search, 94 articles were eligible for inclusion in the systematic review and meta-analysis. The prevalence rate of cryptosporidiosis by using the random effect model among children, healthy people, and gastroenteritis and immunocompromised patients in Iran was estimated as 3.65, 2.94, 1.29, and 4.54%, respectively. Findings of a phylogenetic analysis inferred by gp60 and 18S ribosomal RNA markers indicated that most of the infection rate belonged to C. parvum (particularly subtype IIaA15G2R1) and C. hominis among understudied groups. The present study is the first systematic review and meta-analysis providing a comprehensive view of the prevalence of human cryptosporidiosis and its related risk factors in Iran. It seems that the awareness of Cryptosporidium prevalence, risk factors, and disease complications may be required for developing effective strategies to prevent infection.     

Diversity of <Emphasis Type="Italic">Cryptosporidium</Emphasis> species occurring in sheep and goat breeds reared in Poland

The aim of this study was molecular identification of Cryptosporidium species and assessment of their prevalence in different breeds of sheep and goat reared in Poland. In addition, the relationship between animal age, breed type, and the frequency of Cryptosporidium infections was determined. Fecal samples from 234 lambs and 105 goat kids aged up to 9 weeks, representing 24 breeds and their cross-breeds were collected from 71 small ruminant farms across Poland. The identification of Cryptosporidium species was performed at the 18 SSU ribosomal RNA (rRNA) and COWP loci followed by subtyping of C. parvum and C. hominis strains at GP60 gene locus. The presence of Cryptosporidium DNA at the 18 SSU rRNA locus was detected in 45/234 (19.2%) lamb feces samples and in 39/105 (37.1%) taken from goats. The following Cryptosporidium species: C. xiaoi, C. bovis, C. ubiquitum, C. parvum, and C. hominis were detected in small ruminants. Infections caused by C. xiaoi were predominant without favoring any tested animal species. Subsequent GP60 subtyping revealed the presence of C. parvum IIaA17G1R1 subtype in sheep and IIdA23G1 subtype in goats. IIdA23G1 subtype was detected in a goat host for the first time. There were no significant differences found in frequency of infections between the age groups (<3 and 3–9 weeks) of lambs (P = 0.14, α > 0.05) or goat kids (P = 0.06, α > 0.05). In addition, there was no correlation observed between the frequency in occurrence of particular parasite species and breed type in relation to native sheep breeds (F = 0.11; P = 0.990 > 0.05). In the case of goats, more breed-related differences in parasite occurrence were found. The results of this study improve our knowledge on the breed-related occurrence of Cryptosporidium infections in the population of small ruminants reared in Poland.     

Prevalence,genotypes, and risk factors of <Emphasis Type="Italic">Enterocytozoon bieneusi</Emphasis> in Asiatic black bear (<Emphasis Type="Italic">Ursus thibetanus</Emphasis>) in Yunnan Province,Southwestern China

Microsporidiosis is an important zoonotic disease, even leading to severe diarrhea. However, no information about prevalence and genotypes of Enterocytozoon bieneusi infection in Asiatic black bears in southwestern China is available. The objectives of the present study were to investigate the prevalence of E. bieneusi and to characterize their genotypes using the nested PCR amplification of the internal transcribed spacer region of the ribosomal RNA gene cluster and multilocus sequence typing (MLST). The overall prevalence of E. bieneusi was 19.75% (80/405) and the rate of E. bieneusi in Xishuangbanna (33.33%) was significantly higher than that in any other regions (Honghe, 17.65%; Dehong, 13.04%; Kunming, 0; P?=?0.01). Sequence analysis revealed that 4 known genotypes (D, n?=?2; SC02, n?=?10; SC01, n?=?5; and CHB1, n?=?4) and 13 novel genotypes (designed MJ1–MJ13) were identified. When 17, 5, 14, and 34 sequences at loci MS1, MS3, MS4, and MS7 via MLST analyses, representing 4, 4, 5, and 10 genotypes, respectively, were completed, one multilocus genotype (MLG novel-ABB1) was identified. This is the first report of E. bieneusi in Asiatic black bear in Yunnan province, Southwestern China. The results indicated the potential zoonotic risk of this parasite through the Asiatic black bear in this region and provided foundation data for preventing and controlling E. bieneusi infection of many other animals and humans in these regions.     

Prevalence and antimicrobial resistance of <Emphasis Type="Italic">Ureaplasma</Emphasis> spp. and <Emphasis Type="Italic">Mycoplasma hominis</Emphasis> isolated from semen samples of infertile men in Shanghai,China from 2011 to 2016

This study was conducted to estimate the prevalence and antimicrobial resistance rate of Ureaplasma spp. and Mycoplasma hominis that were isolated from the semen samples of infertile males in Shanghai, China from 2011 to 2016. A total of 5016 infertile males and 412 healthy male controls were examined. The cultivation, identification, and antimicrobial susceptibilities of Ureaplasma spp. and M. hominis were assessed by using a Mycoplasma IST kit that was performed in parallel to selective solid agar cultivation. The positive rate of genital Mycoplasma infections in infertile men from 2011 to 2016 was 30–55%, which initially decreased during the first four years and then increased in the last two. Two distinct high-risk age ranges of Mycoplasma infections were observed: 26–30 years (37.8%) and 31–35 years (30.7%). Semisynthetic tetracyclines and macrolide antibiotics were the most effective agents against Ureaplasma spp. Among the fluoroquinolones, sparfloxacin and levofloxacin were also effective. Antibiotic resistance of Ureaplasma spp. against tetracyclines and macrolide antibiotics in the last six years did not vary significantly. However, the rate of resistance to fluoroquinolones (except norfloxacin) and spectinomycin decreased in the last two years. The rate of genital Mycoplasma presence in infertile patients between the ages of 26 and 35 years in Shanghai was high. The prevalence of genital Mycoplasma decreased during the first four years and then increased, with a peak in 2016. Doxycycline, minocycline, josamycin, and sparfloxacin can be recommended for first-line empirical treatment of Mycoplasma infections in infertile men in Shanghai, China.     

The roundworm <Emphasis Type="Italic">Strongyloides stercoralis</Emphasis> in children,dogs, and soil inside and outside a segregated settlement in Eastern Slovakia: frequent but hardly detectable parasite

A comparative study was carried out to evaluate the Strongyloides stercoralis infections in children and dogs inside and outside the segregated settlement in Medzev, Eastern Slovakia, and a survey of the soil within the settlement was included. Applying the Koga agar plate (KAP) culture method and microscopy examination of stool samples collected from 60 Roma and 21 nonRoma children, no larvae of S. stercoralis were detected but eggs of three nematodes (Ascaris lumbricoides, Trichuris trichiura, and Enterobius vermicularis) and cysts of two protozoan endoparasites (Giardia duodenalis and Cryptosporidium spp.) were often found. However, immunoenzymatic assay (ELISA) for the evidence of IgG antibodies against S. stercoralis showed 33.3% seroprevalence in Roma children and 23.8% prevalence in children from the majority population, attending the same school. Eosinophilia was regularly present in children with exclusive infection of S. stercoralis (eight cases) as well as in individuals suffering from mixed infections of S. stercoralis and some of the above listed parasites (16 cases); high eosinophil counts sometimes, but not always, occurred in parasitized children lacking S. stercoralis antibodies. A comparison of S. stercoralis in dogs from the settlement (40 dogs) and from a distant dog shelter (20 dogs) did not reveal remarkable differences: the direct microscopy of faecal samples revealed rhabditiform larvae in 13.3% of the dogs from the settlement (4/30) and in 10.0% of the dogs from the shelter (2/20). Out of blood samples collected from the second dog group, 55% of the dogs contained antibodies against S. stercoralis. In the soil collected from 14 various locations within the settlement, S. stercoralis larvae were observed in two samples (14.3%); however, 13 samples (92.9%) were positive for human or dog endoparasites of the genera Ancylostoma, Ascaris, Toxocara, Toxascaris, Trichuris, and Hymenolepis.     

Multilocus phylogenetic analysis of <Emphasis Type="Italic">Cryptosporidium andersoni</Emphasis> (Apicomplexa) isolated from a bactrian camel (<Emphasis Type="Italic">Camelus bactrianus</Emphasis>) in China

This is the first report of cryptosporidiosis in a bactrian camel (Camelus bactrianus) in China. Two Cryptosporidium isolates derived from the same bactrian camel (3-year-old) in November 2005 and April 2006 were characterized using sequence and phylogenetic analysis of the small-subunit rRNA (18S rRNA), 70-kDa heat shock protein (HSP70), actin and Cryptosporidium oocyst wall protein (COWP) genes. The sequences of the 18S rRNA and COWP were identical to all other Cryptosporidium andersoni isolates although minor differences were noticed between the isolates and the USA isolate at the actin locus (99.2% of similarity). The sequence of the HSP70 was identical to the Japanese C. andersoni isolate, with a minor difference from the Australian C. andersoni isolate (99.7% of similarity). Cross-transmission studies demonstrated that the C. andersoni isolates did not infect immunosuppressed or immunocompetent Kun-ming mice, severe combined immunodeficiency mice, and immunosuppressed or immunocompetent calves. Among the C. andersoni isolates reported so far, only isolates from Japan could infect SCID mice. Thus, the C. andersoni isolates from the bactrian camel were biologically similar to most bovine C. andersoni isolates characterized so far, but are different from bovine isolates from Japan.     

Clinical and microbiological characteristics of peritoneal dialysis-related peritonitis caused by <Emphasis Type="Italic">Escherichia coli</Emphasis> in southern Taiwan

Peritonitis is a serious complication and major cause of treatment failure in patients undergoing peritoneal dialysis (PD). Escherichia coli is the major pathogen in extraintestinal Gram-negative infections, including PD-related peritonitis. The outcomes of E. coli peritonitis in PD varied from relatively favorable outcomes to a higher incidence of treatment failure. The aim of this study was to investigate the impact of bacterial virulence and host characteristics on the outcomes of PD-related peritonitis caused by E. coli. From January 2000 to June 2016, a total of 47 episodes of monomicrobial and 10 episodes of polymicrobial E. coli PD-related peritonitis, as well as 89 episodes of monomicrobial Gram-positive (56 Staphylococcus spp. and 33 Streptococcus spp.) PD-related peritonitis cases, were retrospectively enrolled. Clinical features, E. coli bacterial virulence, and outcomes were analyzed. Compared to Streptococcus spp. peritonitis, E. coli peritonitis had a higher peritoneal catheter removal rate (38 versus 12%; P =?0.0115). Compared to the monomicrobial group, patients in polymicrobial group were older and had higher peritoneal catheter removal rate (80 versus 38%; P =?0.0324). Treatment failure of E. coli peritonitis was associated with more polymicrobial peritonitis and immunocompromised comorbidity, longer duration of PD therapy, and more antimicrobial resistance. E. coli isolates with more iron-related genes had higher prevalence of phylogenetic group B2 and papG II, iha, ompT, and usp genes. This study demonstrates the important roles of clinical and bacterial characteristics in the outcomes of monomicrobial and polymicrobial E. coli PD-related peritonitis.     

Molecular characterization of <Emphasis Type="Italic">Hepatozoon canis</Emphasis> from farm dogs in Pakistan

Hepatozoon canis is a tick-borne pathogen of canids, which is distributed worldwide. However, very little is known about this protozoan parasite in Pakistan. This study provides the first molecular evidence of H. canis from farm dogs from three agro-ecological zones of Punjab, Pakistan. A conventional PCR targeting the 18S rRNA gene was used to characterize H. canis from farm dogs from three districts, namely Kasur, Rawalpindi, and Muzaffargarh, in Punjab. Of 341 blood samples tested, 155 (45.5%) were positive for H. canis, 73 (61.3%) from Kasur, 46 (42.5%) from Rawalpindi, and 36 (31.5%) from Muzaffargarh. Phylogenetic analyses revealed that 18S rRNA sequences of H. canis from this study clustered in three clades with those of H. canis from previously published studies to the exclusion of all other Hepatozoon spp. included in the analysis. This study provides the first insight into H. canis from farm dogs in Pakistan. Furthermore, it lays a foundation for future studies of the parasite to assess the impact of canine hepatozoonosis in dogs from various agro-ecological zones in Pakistan where pet ownership of dogs is increasing.     

Identification of genus <Emphasis Type="Italic">Campylobacter</Emphasis> up to species level using internal features of 16S rRNA gene sequences

Introduction: 16S rRNA sequencing of novel isolates is one of the preliminary steps in characterization of bacteria, especially when the isolates are of medical relevance. The genus Campylobacter belongs to Class ε-proteobacteria under the Phylum Proteobacteria. It represents economically important species which are gastrointestinal pathogens in humans and livestock animals. Currently, more than 400 16S rDNA sequences belonging to 28 species of this genus are present in the RDP database. But heterogeneity has led to the misplacement of many of these sequences within wrong species. Also, various sequences belonging to Campylobacter have been deposited as orphans. The current study aimed to explore the internal features of 16S rRNA gene sequences in order to develop methods for identification of Campylobacter up to species level. Methods: 428 16S rRNA sequences of 28 species of Campylobacter were analyzed. 392 sequences (>1200 nucleotides, nts) of 16 species were considered for (i) phylogenetic framework analysis and (ii) in silico restriction digestion. 28 uncharacterized sequences present in the database were also investigated in the present study. Results: Phylogenetic framework analysis allowed the identification of genetic variability within Campylobacter species and helped to segregate certain uncharacterized sequences up to species level. 12 out of the 16 species under study showed homogenous behavior, but heterogeneity was observed between C. jejuni and C. coli and C. helveticus and C. upsaliensis respectively. Unique restriction enzymes were identified for six species. Conclusions: The present approach clearly showed that internal features of 16S rRNA is a useful tool for characterization of novel isolates up to the species level. Studies have revealed that niche overlap and consequent increase in the horizontal gene transfer between C. coli and C. jejuni, due to anthropogenic factors, maybe the reason for their heterogeneous nature. This explains the difficulties faced in segregation of the members of these species. 16S rRNA gene proved to be a viable and excellent marker for characterizing the uncharacterized Campylobacter strains leading to a significant diminution in database redundancy. Further, the approaches used in the study might assist in easier identification of the various Campylobacter sequences present in the database.     

<Emphasis Type="Italic">Rickettsia</Emphasis> species in fleas collected from small mammals in Slovakia

Epidemiological and epizootiological studies of Rickettsia felis and other Rickettsia spp. are very important, because their natural cycle has not yet been established completely. In total, 315 fleas (Siphonaptera) of 11 species of Ceratophyllidae, Hystrichopsyllidae and Leptopsyllidae families were tested for the presence of Rickettsia species and Coxiella burnetii with conventional and specific quantitative real-time PCR assays. Fleas were collected from five rodent hosts (Myodes glareolus, Apodemus flavicollis, Apodemus agrarius, Microtus subterraneus, Microtus arvalis) and three shrew species (Sorex araneus, Neomys fodiens, Crocidura suaveolens) captured in Eastern and Southern Slovakia. Overall, Rickettsia spp. was found in 10.8 % (34/315) of the tested fleas of Ctenophthalmus agyrtes, Ctenophthalmus solutus, Ctenophthalmus uncinatus and Nosopsyllus fasciatus species. Infected fleas were coming from A. flavicollis, A. agrarius, and M. glareolus captured in Eastern Slovakia. C. burnetii was not found in any fleas. R. felis, Rickettsia helvetica, unidentified Rickettsia, and rickettsial endosymbionts were identified in fleas infesting small mammals in the Ko?ice region, Eastern Slovakia. This study is the first report of R. felis infection in C. solutus male flea collected from A. agrarius in Slovakia.     

Clonal diversity of <Emphasis Type="Italic">Ureaplasma</Emphasis> species and its relationship with oligozoospermia and semen quality in Chinese infertile males

Whether Ureaplasma spp. are a causative agent of male infertility remains controversial. Previous studies concerning Ureaplasma spp. and male infertility have been confined to the species level of Ureaplasma. Currently, an expanded multilocus sequence typing (eMLST) scheme has been established with high discriminatory power. The aim of this study was to use eMLST to explore the distribution of Ureaplasma spp. and to analyze its role in oligozoospermia and semen quality. A total of 480 semen samples were obtained from Chinese infertile males. The associations between Ureaplasma spp. with oligozoospermia and semen characteristics were further evaluated. Phylogenetic analysis revealed that 102 Ureaplasma spp. could be separated into two clusters and seven sub-groups. Within cluster I (U. parvum), eST16 and eST41 were the most frequent clones. For cluster II (U. urealyticum), eST82 and eST147 were the most prevalent clones. Sub-groups A and C belonging to cluster I and sub-group 1 belonging to cluster II showed an association with oligozoospermia, in contrast with the Ureaplasma spp. negative group (P <?0.05). Compared with the negative group, semen motility decreased in sub-group 2, especially for non-progressive motility (P <?0.05). These results indicated that sub-groups A and C belonging to cluster I (U. parvum) and sub-group 1 belonging to cluster II (U. urealyticum) were shown to be associated with oligozoospermia. Sub-group 2 belonging to cluster II may have the ability to impair semen motility, especially for non-progressive motility.     

Subtype Analysis of Cryptosporidium Specimens from Sporadic Cases in Colorado,Idaho, New Mexico,and Iowa in 2007: Widespread Occurrence of One Cryptosporidium hominis Subtype and Case History of an Infection with the Cryptosporidium Horse Genotype

Subtyping was conducted in late 2007 on 57 Cryptosporidium specimens from sporadic cases in Colorado, Idaho, New Mexico, and Iowa. One previously rare Cryptosporidium hominis subtype was indentified in 40 cases (70%) from all four states, and the Cryptosporidium horse genotype was identified in a pet shop employee with severe clinical symptoms.The protozoan pathogen Cryptosporidium continues to be an important cause of waterborne disease outbreaks and gastrointestinal illness in the United States (4, 6, 21-23). While few recent cryptosporidiosis outbreaks have been associated with drinking water, the incidence of recreational water-associated outbreaks is increasing. In the United States, specimens from patients with outbreak-related cases have been extensively characterized by molecular epidemiology methods (20), but few specimens from patients with sporadic disease have been similarly analyzed (5). As a result, U.S. endemic transmission of Cryptosporidium spp. is not well understood. Conversely, genotyping of a large number of specimens from sporadic cases has facilitated understanding of endemic cryptosporidiosis transmission in the United Kingdom (3, 9, 10).In this study, we genotyped and subtyped specimens from patients with sporadic cryptosporidiosis detected in Colorado, Idaho, New Mexico, and Iowa during the late summer and fall of 2007. The former three states border Utah, where a statewide outbreak of cryptosporidiosis involving 1,902 laboratory-confirmed cases occurred during June to December 2007 (13). Nevertheless, investigations of the sporadic Colorado, Idaho, New Mexico, and Iowa cases revealed no epidemiologic links with any known outbreaks of cryptosporidiosis. These states, and the United States as a whole, reported increased numbers of sporadic cryptosporidiosis cases during 2007 (2). We also report on a symptomatic case of human infection with the Cryptosporidium horse genotype.     

Cervine genotype is the major Cryptosporidium genotype in sheep in China

To identify Cryptosporidium species/genotypes in sheep in China and to elucidate the endemic transmission of cryptosporidiosis, a total of 1,701 fecal samples from five farms in four prefectures in Henan Province (central China) were examined. Eighty-two Cryptosporidium-positive samples were analyzed by polymerase chain reaction (PCR)–restriction fragment length polymorphism analysis of the small subunit (SSU) rRNA gene and PCR analysis of the 60 kDa glycoprotein (gp60) gene, and 41 were further analyzed by DNA sequencing of the PCR products. The SSU rRNA-based PCR identified two Cryptosporidium species and one genotype, including the Cryptosporidium cervine genotype (74/82), Cryptosporidium andersoni (4/82), and Cryptosporidium xiaoi (4/82). The cervine genotype was found in all age groups, C. xiaoi in lambs, and C. andersoni in ewes. There were intragenetic differences in the SSU rRNA gene sequences of the Cryptosporidium cervine genotype and C. xiaoi. No Cryptosporidium parvum was detected by both SSU rRNA- and gp60-based PCR assays. These findings suggest that sheep are a potential source for zoonotic infections of the Cryptosporidium cervine genotype.     

Molecular identification and distribution of Cryptosporidium and Giardia duodenalis in raw urban wastewater in Harbin, China

Contamination of the water supply by protozoa often causes outbreaks of cryptosporidiosis and giardiasis. The goals of the present study was to investigate the level of Cryptosporidium and Giardia duodenalis in wastewater from wastewater treatment plants in Harbin, China, and to understand the endemic transmission characteristics of cryptosporidiosis and giardiasis. Forty-eight domestic wastewater specimens from the two wastewater treatment plants in Harbin City were collected from April 2009 to March 2010. Cryptosporidium spp. and G. duodenalis assemblages were identified by PCR and sequencing of the 18S ribosomal RNA and the triosephosphate isomerase genes, respectively. In total, 15 wastewater specimens were PCR positive for Cryptosporidium and 23 were PCR positive for G. duodenalis. The prevalence of contamination with G. duodenalis (47.9%) was higher than that of Cryptosporidium (31.3%). Molecular identification showed the presence of two Cryptosporidium spp. (14 belonging to Cryptosporidium andersoni and one belonging to Cryptosporidium ubiquitum) and two G. duodenalis assemblages (18 belonging to assemblage AII and six belonging to assemblage B). In addition, eight specimens contained both Cryptosporidium and G. duodenalis, and one specimen contained G. duodenalis assemblages AII and B. These results suggested humans might be the primary source of G. duodenalis contamination in wastewater in the studied area. In contrast, a low prevalence of C. ubiquitum suggested a reduced risk of human cryptosporidiosis caused by C. ubiquitum via waterborne route. This work provides basic experimental data needed for local wastewater treatment plants to develop protective strategies for water safety and to eliminate waterborne parasites.     

Serological reactivity to <Emphasis Type="Italic">Anaplasma phagocytophilum</Emphasis> in neoehrlichiosis patients

The tick-borne bacterium Candidatus (Ca.) Neoehrlichia (N.) mikurensis is a cause of “fever of unknown origin” because this strict intracellular pathogen escapes detection by routine blood cultures. Case reports suggest that neoehrlichiosis patients may display serological reactivity to Anaplasma (A.) phagocytophilum. Since Anaplasma serology is part of the diagnostic work-up of undetermined fever in European tick-exposed patients, we wanted to investigate (1) the prevalence of A. phagocytophilum seropositivity among neoehrlichiosis patients, (2) the frequency of misdiagnosed neoehrlichiosis patients among A. phagocytophilum seropositive patients, and (3) the frequency of A. phagocytophilum and Ca. N. mikurensis co-infections. Neoehrlichiosis patients (n?=?18) were analyzed for A. phagocytophilum IgM and IgG serum antibodies by indirect immunofluorescence assay. Serum samples from suspected anaplasmosis patients (n?=?101) were analyzed for bacterial DNA contents by singleplex PCR specific for A. phagocytophilum and Ca. N. mikurensis, respectively. One fifth of the neoehrlichiosis patients (4/18) were seropositive for IgM and/or IgG to A. phagocytophilum at the time of diagnosis. Among the patients with suspected anaplasmosis, 2% (2/101) were positive for Ca. N. mikurensis by PCR whereas none (0/101) had detectable A. phagocytophilum DNA in the serum. To conclude, patients with suspected anaplasmosis may in fact have neoehrlichiosis. We found no evidence of A. phagocytophilum and Ca. N. mikurensis co-infections in humans with suspected anaplasmosis or confirmed neoehrlichiosis.     

Molecular characterization of arthropod-borne hematozoans in wild mammals from Brazil,Venezuela and Spain

A survey of Babesia, Theileria and Hepatozoon was conducted in wild mammals, including the capybara (Hydrochaeris hydrochaeris; n = 14) from Brazil, the jaguar (Panthera onca; n = 2) and crab-eating raccoon (Procyon cancrivorus; n = 4) from Venezuela, and the red deer (Cervus elaphus; n = 70), red squirrel (Sciurus vulgaris; n = 5) and Eurasian pine marten (Martes martes; n = 3) from Spain. Diagnostic procedures included both microscopy and molecular methods (PCR and sequencing of the 18S rRNA gene). Microscopic examination of blood smears revealed no hematozoan infections — unlike the molecular analyses. Nine Brazilian capybaras were found to be infected with Hepatozoon canis (prevalence 64%), two of which were coinfected with a previously unknown babesid (prevalence 14%) loosely related to Theileria equi (90% 18S rRNA gene similarity according to BLAS® analysis). One jaguar and one crab-eating raccoon from Venezuela were infected by H. canis. Four of the red deer were infected with theilerids (5.7% prevalence), two with Theileria sp. and two with T. annulata. One red squirrel and three pine martens were infected with Hepatozoon sp. The isolate form the red squirrel was phylogenetically related to Hepatozoon sp. reported in Spanish bank voles, whereas those infecting the pine martens were related to Hepatozoon felis reported in Spanish cats. In conclusion, the molecular findings show that some non-canid mammals are carriers of H. canis in South America, while red deer may carry T. annulata in Europe. Small mammals in Europe appear to be unlikely hosts of H. canis and H. felis.     

Parasitic copepods from reef grunts (Teleostei,Haemulidae) with description of a new species of <Emphasis Type="Italic">Lernanthropus</Emphasis> (Siphonostomatoida,Lernanthropidae) from the Mexican Caribbean

During a parasitological survey of the reef ichthyofauna in the Caribbean coast of Mexico, parasitic siphonostomatoid copepods were obtained from the white grunt Haemulon plumierii (Lacepède) and the blue striped grunt H. sciurus (Shaw). Caligus haemulonis Krøyer, 1863 has been recorded from both species of Haemulon in the Northwestern Tropical Atlantic, but it is reported herein for the first time from Mexico. The prevalence and intensity of infection of C. haemulonis has not previously been evaluated in the Caribbean grunts; during this survey, prevalence was highest in H. sciurus; values are similar to those found in other haemulids. Lernanthropus chacchi sp. nov. can be distinguished from its closest congener, L. rathbuni Wilson, 1922, by its female having a dorsal plate covering the entire urosome and the lack of lateral notches at the base of the modified third legs, the male has relatively longer third legs, different body proportions and ornamentation of the first and second legs. This is the third species of Lernanthropus known from reef grunts in the Caribbean and the second species of the genus described from Mexican waters. The infection prevalence of the new species on H. sciurus (24%) was higher than that known for L. rathbuni from other haemulids. Taxonomic illustrations of females and males of both species are also provided. Other crustacean ectoparasites included the copepod Hatschekia spp. and praniza larvae of a gnathid isopod.     

Molecular detection of common intestinal parasites: a performance evaluation of the BD Max™ Enteric Parasite Panel

The purpose of this study was to evaluate the level of agreement of the BD Max? Enteric Parasite Panel (EPP) with microscopy for the detection of Giardia duodenalis, Cryptosporidium spp. and Entamoeba histolytica in stool samples. A total of 372 stool samples (partly collected on the basis of positive microscopy and partly unselected, consecutive sample submitted for parasite investigation) were tested with EPP according to manufacturer’s instructions and also using microscopy according to standard techniques. Discrepant samples were further tested using PCR by the National Parasitology reference laboratory. Levels of agreement and laboratory turnaround times were measured and compared. Overall, positive and negative percent agreement was high between the two methods. However, microscopy resulted in four false positives and one false negative for G. duodenalis and two false positives for Cryptosporidium. Additionally, microscopy could not differentiate between E. histolytica and Entamoeba dispar. Median laboratory turnaround time was 65 hours for microscopy; results from EPP could be available after four hours. Blastocycstis hominis was detected by microscopy in one sample and would have been missed if only EPP was performed. The EPP was a good alternative to microscopy, detecting a small number of additional positives that were missed by microscopy. The assay is significantly faster than microscopy and allows laboratory workflows to be streamlined. The risk of missing parasites that are not included in the EPP appears to be minimal in the studied population; however, there may be certain patient groups who would benefit from microscopic examination of stools.     

Prevalence and distribution of Cryptosporidium spp. in dairy cattle in Heilongjiang Province, China

Few data are available on the molecular characterization of Cryptosporidium spp. in cattle in China. In the present study, a total of 507 fecal specimens from six dairy farms in Heilongjiang Province were examined for Cryptosporidium spp. by light microscopy of concentrates from the formalin-ethyl acetate sedimentation method (for less than 2-month-old calves) or Sheather’s floatation method (more than 3-month-old dairy cattle). Twenty-seven post-weaned calves on five farms were positive for Cryptosporidium oocysts. PCR and DNA sequence analysis of the 18S rRNA, actin, and 70 kDa heat shock protein genes identified Cryptosporidium andersoni and Cryptosporidium. ryanae, with C. andersoni as the dominant species (26 out of 27). In comparison with other regions of the world, the distribution of Cryptosporidium species in the areas appears to be unique.