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rapid and sensitive detection of babesia bovis and babesia bigemina by loop-mediated isothermal amplification combined with a lateral flow dipstick.babesia spp. are apicomplexan protozoan parasites of the red blood cells of mammals and are transmitted by ticks. bovine babesiosis mainly caused by babesia bovis and babesia bigemina occurs worldwide, which is a great threat to animal health. microscopy examination is a gold standard for the diagnosis of babesiosis. however, its sensitivity is too low. this study was conducted to establish a simple, efficient and fast lamp-lfp method used for early diagnosis of animal babesiosis. lamp was devel ...201626921043
replication and maintenance of the plasmodium falciparum apicoplast genome.members of the phylum apicomplexa are responsible for many devastating diseases including malaria (plasmodium spp.), toxoplasmosis (toxoplasma gondii), babesiosis (babesia bovis), and cyclosporiasis (cyclospora cayetanensis). most apicomplexans contain a unique and essential organelle called the apicoplast. derived from an ancient chloroplast, the apicoplast replicates and maintains a 35 kilobase (kb) circular genome. due to its essential nature within the parasite, drugs targeted to proteins in ...201627338018
molecular survey of zoonotic agents in rodents and other small mammals in croatia.croatia is a focus for many rodent-borne zoonosis. here, we report a survey of 242 rodents and small mammals, including 43 myodes glareolus, 131 apodemus flavicollis, 53 apodemus agrarius, three apodemus sylvaticus, six sorex araneus, four microtus arvalis, one microtus agrestis, and one muscardinus avellanarius, collected at eight sites in croatia over an 8-year period. multiplex masstag polymerase chain reaction (pcr) was used for detection of borrelia, rickettsia, bartonella, babesia, ehrlich ...201626711522
molecular phylogeny of toxoplasmatinae: comparison between inferences based on mitochondrial and apicoplast genetic sequences.phylogenies within toxoplasmatinae have been widely investigated with different molecular markers. here, we studied molecular phylogenies of the toxoplasmatinae subfamily based on apicoplast and mitochondrial genes. partial sequences of apicoplast genes coding for caseinolytic protease (clpc) and beta subunit of rna polymerase (rpob), and mitochondrial gene coding for cytochrome b (cytb) were analyzed. laboratory-adapted strains of the closely related parasites sarcocystis falcatula and sarcocys ...201627007245
besnoitia besnoiti lytic cycle in vitro and differences in invasion and intracellular proliferation among isolates.bovine besnoitiosis, caused by the protozoan besnoitia besnoiti, reduces productivity and fertility of affected herds. besnoitiosis continues to expand in europe and no effective control tools are currently available. experimental models are urgently needed. herein, we describe for the first time the kinetics of standardised in vitro models for the b. besnoiti lytic cycle. this will aid to study the pathogenesis of the disease, in the screening for vaccine targets and drugs potentially useful fo ...201626928430
characterization of an igg monoclonal antibody targeted to both tissue cyst and sporocyst walls of toxoplasma gondii.toxoplasma gondii infects animals habiting terrestrial and aquatic environments. its oocysts and tissue cysts are important for the horizontal transmission of this parasite. the oocyst and tissue cyst walls are crucial for the ability of the parasite to persist in the environment or in animal tissues, respectively. however, the composition of these walls is not well understood. we report the generation of monoclonal antibodies directed against wall components using mice immunized with oocyst ant ...201626836446
naturally acquired bovine besnoitiosis: differential distribution of parasites in the skin of chronically infected cattle.bovine besnoitiosis is caused by besnoitia besnoiti, an apicomplexan parasite closely related to toxoplasma gondii and neospora caninum. in the acute stage of besnoitiosis, cattle suffer from pyrexia, swollen lymph nodes, anorexia and subcutaneous edema. in the chronic stage, tissue cysts are formed in a variety of tissues including the skin. knowledge about the distribution of tissue cysts of different parts of the skin of infected animals is scarce. four chronically infected cattle were euthan ...201626801602
endoparasite survey of free-swimming baleen whales (balaenoptera musculus, b. physalus, b. borealis) and sperm whales (physeter macrocephalus) using non/minimally invasive methods.a number of parasitic diseases have gained importance as neozoan opportunistic infections in the marine environment. here, we report on the gastrointestinal endoparasite fauna of three baleen whale species and one toothed whale: blue (balaenoptera musculus), fin (balaenoptera physalus), and sei whales (balaenoptera borealis) and sperm whales (physeter macrocephalus) from the azores islands, portugal. in total, 17 individual whale fecal samples [n = 10 (b. physalus); n = 4 (p. macrocephalus); n = ...201626593736
prevalence of leishmania infantum and co-infections in stray cats in northern italy.stray cats in the city of milan, italy, were tested for leishmania infantum and other selected infections. twenty-seven cats (30.0%) were seroreactive by indirect fluorescent antibody test (ifat), with an antibody titer of 1:40 for 16 (17.7%) cats and 1:80 (cut-off for feline l. infantum infection) for 11 (12.2%) cats. one blood (1.1%) and one popliteal lymph node (1.1%) sample tested positive by real-time polymerase chain reaction; no oculoconjunctival swabs tested positive. feline immunodefici ...201627012922
cat scratches, not bites, are associated with unipolar depression--cross-sectional study.a recent study performed on 1.3 million patients showed a strong association between being bitten by a cat and probability of being diagnosed with depression. authors suggested that infection with cat parasite toxoplasma could be the reason for this association.201626728406
simultaneous multiview capture and fusion improves spatial resolution in wide-field and light-sheet microscopy.most fluorescence microscopes are inefficient, collecting only a small fraction of the emitted light at any instant. besides wasting valuable signal, this inefficiency also reduces spatial resolution and causes imaging volumes to exhibit significant resolution anisotropy. we describe microscopic and computational techniques that address these problems by simultaneously capturing and subsequently fusing and deconvolving multiple specimen views. unlike previous methods that serially capture multip ...201627761486
cd19 lymphocyte proliferation induced by bifidobacterium animalis subsp. lactis in c57bl/6 mice experimentally infected with toxoplasma gondii.toxoplasmosis is frequently acquired through the oral route by the ingestion of cysts or oocysts of toxoplasma gondii. once ingested, the parasites penetrate the intestinal epithelial cells and rapidly disseminate to all organs in the host. during t. gondii infection, the intestinal microbiota plays an important role in stimulating a protective immune response against the parasite. in this sense the use of probiotics is worthy of note since they are live microorganisms that have beneficial effec ...201627074320
neglected waterborne parasitic protozoa and their detection in water.outbreak incidents raise the question of whether the less frequent aetiological agents of outbreaks are really less frequent in water. alternatively, waterborne transmission could be relevant, but the lack of attention and rapid, sensitive methods to recover and detect the exogenous stages in water may keep them under-recognized. high quality information on the prevalence and detection of less frequent waterborne protozoa, such as cyclospora cayetanensis, toxoplasma gondii, isospora belli, balan ...201627281375
molecular mimicry and autoimmune thyroid disease.hypothesized 40 years ago, molecular mimicry has been thereafter demonstrated as an extremely common mechanism by which microbes elude immune response and modulate biosynthetic/metabolic pathways of the host. in genetically predisposed persons and under particular conditions, molecular mimicry between microbial and human antigens can turn a defensive immune response into autoimmunity. such triggering role and its pathogenetic importance have been investigated and demonstrated for many autoimmune ...201627307072
prevalence of antibodies to selected viruses and parasites in introduced and endemic carnivores in western madagascar.introduced animals impact endemic populations through predation, competition, and disease transmission. populations of endemic carnivores in madagascar are declining, and pathogens transmitted from introduced species may further endanger these unique species. we assessed the exposure of introduced and endemic carnivores to common viral and parasitic pathogens in two national parks of madagascar (kirindy mitea national park and ankarafantsika national park) and their neighboring villages. we also ...201627195685
prevalence of antibodies for selected canine pathogens among wolves (canis lupus) from the alaska peninsula, usa.we collected blood samples from wolves ( canis lupus ) on the alaska peninsula, southwest alaska, us, 2006-11 and tested sera for antibodies to canine adenovirus (cav), canine coronavirus (ccv), canine distemper virus (cdv), canine herpesvirus (chv), canine parainfluenza (cpi), canine parvovirus (cpv), neospora caninum , and toxoplasma gondii . detected antibody prevalence was 90% for cav, 28% for ccv, 12% for cdv, 93% for chv, 0% for cpi, 20% for cpv, 0% for n. caninum, and 86% for t. gondii . ...201627195683
knowledge gaps in host-parasite interaction preclude accurate assessment of meat-borne exposure to toxoplasma gondii.toxoplasma gondii is recognized as a widely prevalent zoonotic parasite worldwide. although several studies clearly identified meat products as an important source of t. gondii infections in humans, quantitative understanding of the risk posed to humans through the food chain is surprisingly scant. while probabilistic risk assessments for pathogens such as campylobacter jejuni, listeria monocytogenes or escherichia coli have been well established, attempts to quantify the probability of human ex ...201628139250
developing a salivary antibody multiplex immunoassay to measure human exposure to environmental pathogens.the etiology and impacts of human exposure to environmental pathogens are of major concern worldwide and, thus, the ability to assess exposure and infections using cost effective, high-throughput approaches would be indispensable. this manuscript describes the development and analysis of a bead-based multiplex immunoassay capable of measuring the presence of antibodies in human saliva to multiple pathogens simultaneously. saliva is particularly attractive in this application because it is noninv ...201627685162
safety and efficacy of intravitreal dexamethasone implants in the management of macular edema secondary to infectious uveitis.to assess the safety and efficacy of intravitreal dexamethasone implants in the treatment of macular edema secondary to infectious uveitis.201627183031
molecular assessment of the transplacental transmission of toxoplasma gondii, neospora caninum, brucella canis and ehrlichia canis in dogs.given the fact that numerous microbial species can be detected in pregnant female dogs, the objective of this study was to assess the transplacental transmission of brucella canis, ehrlichia canis, neospora caninum and toxoplasma gondii in stillborn puppies. this study involved 41 stillborn puppies, 78.6% of which were positive for t. gondii, 52.4% for n. caninum and 59.5% for b. canis. e. canis was not detected in any of the analyzed puppies. pregnancy is an important physiological condition fo ...201627865263
detection and sequencing of zika virus from amniotic fluid of fetuses with microcephaly in brazil: a case study.the incidence of microcephaly in brazil in 2015 was 20 times higher than in previous years. congenital microcephaly is associated with genetic factors and several causative agents. epidemiological data suggest that microcephaly cases in brazil might be associated with the introduction of zika virus. we aimed to detect and sequence the zika virus genome in amniotic fluid samples of two pregnant women in brazil whose fetuses were diagnosed with microcephaly.201626897108
recombinant canine adenovirus type-2 expressing tgrop16 provides partial protection against acute toxoplasma gondii infection in mice.we previously demonstrated that the survival time of balb/c mice challenged with toxoplasma gondii rh strain was prolonged by immunising the mice with a eukaryotic vector expressing the protein rop16 of t. gondii. building upon previous findings, we are exploring improved vaccination strategies to enhance protection. in this work, a novel recombinant canine adenovirus type 2 expressing rop16 (cav-2-rop16) of t. gondii was constructed and identified to express rop16 in madin-darby canine kidney c ...201627742446
first report of toxoplasma gondii, dirofilaria immitis, and chlamydia felis infection in stray and companion cats in northeastern and eastern china.feline vector-borne diseases have increasingly become a focus of interest in recent years. toxoplasma gondii, dirofilaria immitis, and chlamydia felis are common pathogens of cats that can affect humans among different countries all over the world. however, information about prevalence of t. gondii and c. felis is available in china, but information about coinfection of t. gondii, d. immitis, and c. felis in cats is limited. thus, this study was conducted to estimate the prevalence of these path ...201627463689
infectious disease and toxicological monitoring of stranded pacific harbor seals (phoca vitulina richardsi) in cook inlet as surrogates for monitoring endangered belugas (delphinapterus leucas).pacific harbor seals ( phoca vitulina richardsi) and belugas ( delphinapterus leucas ) eat many of the same prey species, occupy the same geographic area, and demonstrate site fidelity in cook inlet, alaska. although most direct research involving the critically endangered belugas is currently prohibited, studying harbor seals may provide important information about this beluga population. in recent years, harbor seal populations in alaska have declined for unknown reasons. as part of its strand ...201627691941
post-mortem findings in southern right whales eubalaena australis at península valdés, argentina, 2003-2012.between 2003 and 2012, 605 southern right whales (srw; eubalaena australis) were found dead along the shores of península valdés (pv), argentina. these deaths included alarmingly high annual losses between 2007 and 2012, a peak number of deaths (116) in 2012, and a significant number of deaths across years in calves-of-the-year (544 of 605 [89.9%]; average = 60.4 yr(-1)). post-mortem examination and pathogen testing were performed on 212 whales; 208 (98.1%) were calves-of-the-year and 48.0% of t ...201627068500
blindness in a wild american black bear cub (ursus americanus).an approximately six-month-old wild american black bear (ursus americanus) was found wandering in saskatchewan and was presented to the veterinary medical centre of the western college of veterinary medicine for apparent blindness. clinical examination confirmed an inability to navigate a photopic maze, bilateral tapetal hyper-reflectivity, fundi devoid of retinal vessels, and small pale optic nerve papillae. single-flash electroretinography revealed a and b-wave amplitudes of approximately 40 a ...201626302466
investigation of a mortality cluster in wild adult yellow-eyed penguins (megadyptes antipodes) at otago peninsula, new zealand.we investigated an epidemic mortality cluster of yellow-eyed penguins (megadyptes antipodes) that involved 67 moribund or dead birds found on various beaches of the otago peninsula, new zealand, between january 21 and march 20, 2013. twenty-four carcases were examined postmortem. histological lesions of pulmonary, hepatic and splenic erythrophagocytosis and haemosiderosis were found in 23 of 24 birds. fifteen birds also had haemoglobin-like protein droplets within renal tubular epithelial cells. ...201627919180
microbiological and parasitological survey of zoonotic agents in apparently healthy feral pigeons.microbiological and parasitological investigation was carried out on a colony of feral pigeons, located in a green area near the main hospital of a central italy city. one hundred pigeons were submitted to clinical examination. cloacal swabs, grouped in pool of 4 samples, were analyzed to detect the presence of coxiella burnetii, chlamydia psittaci, chlamydophila spp. using a biomolecular procedure, while individual cloacal samples were examined for salmonella spp., campylobacter spp., and yeast ...201627487504
current and future molecular diagnostics for ocular infectious diseases.confirmation of ocular infections can pose great challenges to the clinician. a fundamental limitation is the small amounts of specimen that can be obtained from the eye. molecular diagnostics can circumvent this limitation and have been shown to be more sensitive than conventional culture. the purpose of this review is to describe new molecular methods and to discuss the applications of next-generation sequencing-based approaches in the diagnosis of ocular infections.201627585214
[detection of echinococcus granulosus and echinococcus multilocularis in cyst samples using a novel single tube multiplex real-time polymerase chain reaction].cystic echinococcosis (ce) and alveolar echinococcosis (ae) caused by echinococcus granulosus and echinococcus multilocularis, respectively, are important helminthic diseases worldwide as well as in our country. epidemiological studies conducted in turkey showed that the prevalence of ce is 291-585/100.000. it has also been showed that the seroprevalence of ae is 3.5%. for the diagnosis of ce and ae, radiological (ultrasonography, computed tomography, magnetic resonance) and serological methods, ...201627175499
detection by pcr of pathogenic protozoa in raw and drinkable water samples in colombia.we evaluated the presence of dna of giardia, toxoplasma, and cryptosporidium by pcr, and of giardia and cryptosporidium genera by immunofluorescence antibody test (ifat), in water samples, before, during, and after plant treatment for drinkable water. we applied this method in 38 samples of 10 l of water taken from each of the water treatment steps and in 8 samples taken at home (only for toxoplasma pcr) in quindio region in colombia. there were 8 positive samples for cryptosporidium parvum (21  ...201626779921
novel bumped kinase inhibitors are safe and effective therapeutics in the calf clinical model for cryptosporidiosis.cryptosporidiosis, caused by the apicomplexan parasite cryptosporidium parvum, is a diarrheal disease that has produced a large global burden in mortality and morbidity in humans and livestock. there are currently no consistently effective parasite-specific pharmaceuticals available for this disease. bumped kinase inhibitors (bkis) specific for parasite calcium-dependent protein kinases (cdpks) have been shown to reduce infection in several parasites having medical and veterinary importance, inc ...201627923949
bioaccumulation of toxoplasma and cryptosporidium by the freshwater crustacean gammarus fossarum: involvement in biomonitoring surveys and trophic transfer.the protozoa toxoplasma gondii and cryptosporidium parvum are public health priorities because their oocysts can persist in recreational, surface, drinking, river, and sea water sources for a long time. to evaluate the capacity of the freshwater crustacean gammarus fossarum to accumulate t. gondii and c. parvum oocysts, gammarids were exposed to 200, 2000 or 20,000 oocysts per gammarid and per day for 21 days followed by 5 days of depuration. c. parvum dna was detected by qpcr in g. fossarum in ...201627454203
assessment and molecular characterization of human intestinal parasites in bivalves from orchard beach, ny, usa.bivalves have been shown to be carriers of the human intestinal parasites cryptosporidium parvum and toxoplasma gondii. the goal of this study is to determine the prevalence of protozoan parasites in mollusks of new york city using a polymerase chain reaction (pcr)-based assay. four species of mollusks, mya arenaria, geukensia demissa, crassostrea virginica, and mytilis edulis, were collected from orchard beach, ny in the fall of 2014, totaling 159 specimens. each individual mollusk was dissecte ...201627043590
drug repurposing: mining protozoan proteomes for targets of known bioactive compounds.to identify potential opportunities for drug repurposing by developing an automated approach to pre-screen the predicted proteomes of any organism against databases of known drug targets using only freely available resources.201623757409
characterization of the neospora caninum ncrop40 and ncrop2fam-1 rhoptry proteins during the tachyzoite lytic cycle.virulence factors from the rop2-family have been extensively studied in toxoplasma gondii, but in the closely related neospora caninum only ncrop2fam-1 has been partially characterized to date. ncrop40 is a member of this family and was found to be more abundantly expressed in virulent isolates. both ncrop2fam-1 and ncrop40 were evaluated as vaccine candidates and exerted a synergistic effect in terms of protection against vertical transmission in mouse models, which suggests that they may be re ...201626521890
the tandemly repeated ntpase (ntpdase) from neospora caninum is a canonical dense granule protein whose rna expression, protein secretion and phosphorylation coincides with the tachyzoite egress.ntpases (also ntpdases) are enzymes with apyrase activity. they are widely distributed among eukaryotes, and also among members of the family sarcocystidae. in toxoplasma gondii, the tgntpase accumulates in the dense granules, and has been commonly associated with the strain virulence. in the closely related neospora caninum, the ncntpase lacks nucleoside diphosphate hydrolase activity and appears to be more abundant in virulent isolates, indicating that it may contribute to the pathogenicity of ...201627329357
toxoplasma gondii and neospora caninum in free-range chickens in henan province of china.background. chickens serve as an intermediate host for toxoplasma gondii and neospora caninum; infection of free-range (fr) chickens with these organisms is a useful indicator of soil and environmental contamination with oocysts. a total of 700 fr chicken serum samples and 300 heart samples were collected from henan province from march to july 2015. antibodies to t. gondii were found in 18.86% (132/700) of the chickens by modified agglutination test (cut-off 1 : 25), while 23.14% (162/700) were ...201627274992
pathology, immunohistochemistry, and ultrastructural findings associated with neurological sarcocystosis in cattle.paraffin-embedded blocks of brain of a nine months old bull calf that died of neurological signs in 1982 in germany were restudied. numerous schizonts and merozoites were found associated with extensive but focal necrosis and severe meningoencephalitis. developing stages of schizonts as well as free merozoites were identified. the schizonts were primarily in perivascular areas. ultrastructurally, schizonts were seen both in capillaries and in extravascular space. merozoites were often concentrat ...201627198793
toxoplasma gondii and neospora caninum in wild small mammals: seroprevalence, dna detection and genotyping.generally, rodents and other small mammals are considered as one of the sources of toxoplasma gondii or neospora caninum infection for cats and dogs as the definitive hosts of these two parasites, respectively. the aim of the study was to find out the prevalence of these two parasites in wild small mammals from the czech republic and to characterize t. gondii isolates by methods of molecular biology. a total of 621 wild small mammals were caught in the czech republic during years 2002-2014. anti ...201627198782
seroprevalence and risk factors of infections with neospora caninum and toxoplasma gondii in hunting dogs from campania region, southern italy.hunting dogs have probably a higher level of exposure to neospora caninum dubey, carpenter, speer, topper et uggla, 1988 and toxoplasma gondii nicolle et manceaux, 1908 than other canine populations for their different lifestyle. the aim of our survey was to determine the seroprevalence of n. caninum and t. gondii in hunting dogs from southern italy and assess risk factors related to these protozoan infections. blood samples were collected from 398 hunting dogs (19 different breeds, aged from 5 ...201627189127
neonatal neosporosis in a 2-week-old bernese mountain dog infected with multiple neospora caninum strains based on ms10 microsatellite analysis.neonatal neosporosis is a challenging disease to diagnose in neonatal and young puppies because the first signs of this condition may not be strongly suggestive of an infectious aetiology. within two weeks of birth, three of four pups died with a subacute clinical course, some with dyspnea, some with diarrhoea and some with neurologic signs. neosporosis was diagnosed post-mortem, but only after microscopic examination of tissues collected at necropsy. histological findings consisted of (i) necro ...201627084485
in vitro and in vivo effects of the phytohormone inhibitor fluridone against neospora caninum infection.neospora caninum causes abortion and stillbirth in cattle. identification of effective drugs against this parasite remains a challenge. previous studies have suggested that disruption of abscisic acid (aba)-mediated signaling in apicomplexan parasites such as toxoplasma gondii offers a new drug target. in this study, the aba inhibitor, fluridone (flu), was evaluated for its action against n. caninum. production of endogenous aba within n. caninum was confirmed by ultra-performance liquid chromat ...201627021922
neospora caninum and toxoplasma gondii antibodies in red foxes (vulpes vulpes) in the czech republic.neospora caninum and toxoplasma gondii are worldwide spread parasites, causing serious illnesses in sensitive animals; toxoplasmosis is also important zoonosis. although neosporosis is not considered as a zoonosis, it leads to aborted births in cattle, as well as paresis and paralysis in dogs.201627007522
anti-toxoplasma gondii and anti-neospora caninum antibodies in capybaras (hydrochoerus hydrochaeris) from itu municipality, são paulo.capybaras (hydrochoerus hydrochaeris) are the largest rodents found in south america. the aim of the present study was to investigate the occurrence of anti-toxoplasma gondii and anti-neospora caninum antibodies in 170 free-living capybaras in a residential park area in itu municipality, são paulo state, brazil. serum samples were tested by indirect fluorescent antibody test (ifat) for t. gondii (ifat ≥ 1:16) and n. caninum (ifat ≥ 1:50). among the 170 samples analyzed, 10% (17/170) and 0% (0/17 ...201626982562
effect of different ecosystems and management practices on toxoplasma gondii and neospora caninum infections in wild ruminants in spain.toxoplasma gondii and neospora caninum are two major abortifacient protozoans in domestic small ruminants and cattle, respectively, and they also parasitize a wide range of wildlife. numerous serosurveys have been conducted in wild ruminants worldwide. however, the potential effect of different ecosystems and management practices on these infections has not been investigated. we studied the prevalence of antibodies to t. gondii and n. caninum in wild ruminants between 2007 and 2012 from four nat ...201626967135
parasitic and bacterial infections of myocastor coypus in a metropolitan area of northwestern italy.coypus (myocastor coypus) are widespread throughout europe. in northern italy, they are abundant in the flatland areas, and their high population densities can cause economic loss and ecosystem damage. we examined 153 coypus for selected parasitic and bacterial infections. we found strongyloides myopotami (63.4% prevalence), trichostrongylus duretteae (28.1%), eimeria coypi (86.3%), and eimeria seideli (6.8%), but did not find giardia duodenalis or cryptosporidium spp. we also isolated staphyloc ...201626745834
cloning, expression and immunoreactivity of recombinant toxoplasma gondii gra5 protein.toxoplasma gondii is an obligatory intracellular parasite which causes severe diseases in the fetus of pregnant women and immunocopmromised patients. serological tests based on recombinant protein are one of the main diagnosis methods for the detection of specific antibodies in serum samples. dense granule antigenic proteins derived from t. gondii (tggras) are potential antigens for the development of diagnostic tools.201628149494
toxoplasma gondii plaque assays revisited: improvements for ultrastructural and quantitative evaluation of lytic parasite growth.lytic growth of intracellular toxoplasma gondii tachyzoite stages over a period of days results in plaques within mononolayers of host cells. plaque assays are in frequent use to isolate single clones and to investigate invasion, replication and egress over a longer time frame. to allow correlating plaque morphology and/or size with ultrastructural examination of individual parasites we introduce a simple protocol for correlative light and electron microscopy (clem) of entire plaques. we also il ...201628011168
evaluation of protective immune responses induced by recombinant trxlp and eno2 proteins against toxoplasma gondii infection in balb/c mice.toxoplasma gondii is an obligate intracellular parasitic protozoan that can infect almost all species of warm-blooded animals. as any chemical-based drugs could not act against the tissue cyst stage of t. gondii, vaccination may be one of the ideal control strategies. in the present study, two new vaccine candidates, named tgeno2 and tgtrxlp, were purified from escherichia coli with pet-30a(+) expression system and then were injected into balb/c mice to evaluate the protective efficacy against a ...201627803923
characterization of aspartyl aminopeptidase from toxoplasma gondii.aminopeptidases have emerged as new promising drug targets for the development of novel anti-parasitic drugs. an aspartyl aminopeptidase-like gene has been identified in the toxoplasma gondii genome (tgaap), although its function remains unknown. in this study, we characterized tgaap and performed functional analysis of the gene product. firstly, we expressed a functional recombinant tgaap (rtgaap) protein in escherichia coli, and found that it required metal ions for activity and showed a subst ...201627678060
immunoreactivity analysis of toxoplasma gondii recombinant antigen rsag3 in sera from immunized balb/c mice and toxoplasmosis patients.the coccidian protozoa toxoplasma gondii is an obligate intracellular parasite of humans and other warm-blooded animals. diagnosis of toxoplasmosis is of considerable medical importance for human, especially pregnant women and immunocompromised individuals. the apply of an escherichia coli recombinant antigen(s) would be significantly useful in developing standardization of the diagnostic tests and reducing their costs. in this study, immunoreactivity of recombinant sag3 against sera from immuni ...201627516998
twenty-six circulating antigens and two novel diagnostic candidate molecules identified in the serum of canines with experimental acute toxoplasmosis.the protozoan toxoplasma gondii is a pathogen that causes severe opportunistic disease in a wide range of hosts. efficient methods to diagnose acute t. gondii infection are essential for the administration of appropriate treatments and to reduce economic losses. in animals with acute infections, circulating antigens (cags) were detected as early as two days post-infection; these cags were reliable diagnostic indicators of acute infection. however, only a limited number of cags have been identifi ...201627357215
functional characterization of x-prolyl aminopeptidase from toxoplasma gondii.in the present study, a recombinant aminopeptidase p (rtgapp) from toxoplasma gondii was expressed in escherichia coli to evaluate its enzyme parameters. the rtgapp showed strong activity against a synthetic substrate for aminopeptidase p at ph 8·0 with a k m value of 0·255 µ m and a k cat value of 35·6 s-1. the overall catalytic efficiency (k cat/k m) of the rtgapp was 139·6 × 105 m-1 s-1. the activity of rtgapp was enhanced by the addition of divalent cations and inhibited by bestatin. deletio ...201627220680
toll-like receptor 11 (tlr11) interacts with flagellin and profilin through disparate mechanisms.toll-like receptors (tlrs) are innate immune receptors that sense a variety of pathogen-associated molecular patterns (pamps) by interacting with them and subsequently initiating signal transduction cascades that elicit immune responses. tlr11 has been shown to interact with two known protein pamps: salmonella and e. coli flagellin flic and toxoplasma gondii profilin-like protein. given the highly divergent biology of these pathogens recognized by tlr11, it is unclear whether common mechanisms a ...201626859749
proteome-wide identification of lysine succinylation in the proteins of tomato (solanum lycopersicum).post-translational modification of proteins through lysine succinylation plays important regulatory roles in living cells. lysine succinylation was recently identified as a novel post-translational modification in escherichia coli, yeast, toxoplasma gondii, hela cells, and mouse liver. interestingly, only a few sites of lysine succinylation have been detected in plants to date. in this study, we identified 347 sites of lysine succinylation in 202 proteins in tomato by using high-resolution mass ...201626828863
protozoan parasites.• stool antigen detection for cryptosporidium sp, giardia lamblia and entamoeba histolytica are now commercially available, have better sensitivity and specificity than the traditional stool microscopy, and are less dependent on personnel skill. tests employing newer techniques with faster turnaround time are also available for diagnosing trichomoniasis.• nitazoxanide, the only u.s. food and drug administration-approved medication for therapy of cryptosporidiosis, is effective among immunocompet ...201626834225
screening and characterization of apical membrane antigen 1 interacting proteins in eimeria tenella.avian coccidiosis is a widespread and economically significant disease of poultry. it is an enteric disease caused by several protozoan eimeria species. eimeria belongs to the phylum apicomplexa, which exhibits an unusual mechanism of host cell invasion. during invasion of host cells, the protein apical membrane antigen 1 (ama1) is essential for invasion of toxoplasma gondii and plasmodium. contrary to the roles of ama1 during host cell invasion in t. gondii and plasmodium, the precise functions ...201627693220
regulation of dna replication proteins in parasitic protozoans: possible role of cdk-like kinases.regulatory roles of cdks in fundamental processes including cell cycle progression and transcription are well conserved in metazoans. this family of proteins has undergone significant evolutionary divergence and specialization. several cdk-like kinases have been identified and characterized in parasitic protozoans. however, clear functional role and physiological relevance of these proteins in protozoans still remain elusive. in continuation with the recent finding that cdk-like protein pfpk5 re ...201626780367
early life socioeconomic position and immune response to persistent infections among elderly latinos.persistent infections, such as cytomegalovirus (cmv), herpes simplex virus-1 (hsv-1), helicobacter pylori (h. pylori), and toxoplasma gondii (t. gondii), are common in the u.s. but their prevalence varies by socioeconomic status. it is unclear if early or later life socioeconomic position (sep) is a more salient driver of disparities in immune control of these infections. using data from the sacramento area latino study on aging, we examined whether early or later life sep was the strongest pred ...201627543684
exposure to perinatal infections and bipolar disorder: a systematic review.bipolar disorder (bd) is a debilitating psychiatric disorder and a growing global public health issue. notwithstanding bd has been conceptualized as a neuroprogressive illness, there are some evidences to suggest a role for neurodevelopmental pathways in the patho-etiology of this disorder. evidences on the associations between perinatal infections and risk for bipolar disorder have been inconsistent across studies. here, we performed a systematic review of observational studies on the relations ...201626812921
evaluation of a standardised real-time pcr based dna-detection method (realstar®) in whole blood for the diagnosis of primary human cytomegalovirus (cmv) infections in immunocompetent patients.cytomegalovirus (cmv) dna detection in blood could, as a supplementary test to serology, improve the accuracy and speed of diagnosis of an acute cmv infection. in this study we evaluated the performance of a commercially available and standardised cmv pcr assay in whole blood for the diagnosis of a primary infection in immunocompetent adults. moreover, the kinetics of viral dna was evaluated in order to provide a time frame in which viral dna could be detected during an acute primary infection. ...201626661089
torch "co-infections" are associated with increased risk of abortion in pregnant women.torch infections (toxoplasmosis, rubella, cytomegalovirus and herpes simplex virus) have long been known to be associated with bad obstetric outcomes. however, little information is available about the impact of torch co-infections on the outcome of pregnancy. hence, we tested the igg and igm antibodies to toxoplasma gondii, rubella, cytomegalovirus and herpes simplex virus among 81 pregnant women with abortion (case group) and 98 pregnant women with normal delivery (control group). in the singl ...201626499091
the e3 ubiquitin ligase adaptor protein skp1 is glycosylated by an evolutionarily conserved pathway that regulates protist growth and development.toxoplasma gondii is a protist parasite of warm-blooded animals that causes disease by proliferating intracellularly in muscle and the central nervous system. previous studies showed that a prolyl 4-hydroxylase related to animal hifα prolyl hydroxylases is required for optimal parasite proliferation, especially at low o2. we also observed that pro-154 of skp1, a subunit of the skp1/cullin-1/f-box protein (scf)-class of e3-ubiquitin ligases, is a natural substrate of this enzyme. in an unrelated ...201626719340
dendrimer-rna nanoparticles generate protective immunity against lethal ebola, h1n1 influenza, and toxoplasma gondii challenges with a single dose.vaccines have had broad medical impact, but existing vaccine technologies and production methods are limited in their ability to respond rapidly to evolving and emerging pathogens, or sudden outbreaks. here, we develop a rapid-response, fully synthetic, single-dose, adjuvant-free dendrimer nanoparticle vaccine platform wherein antigens are encoded by encapsulated mrna replicons. to our knowledge, this system is the first capable of generating protective immunity against a broad spectrum of letha ...201627382155
toxoplasma gondii, dirofilaria immitis, feline immunodeficiency virus (fiv), and feline leukemia virus (felv) infections in stray and pet cats (felis catus) in northwest china: co-infections and risk factors.this study was conducted to estimate the prevalence of toxoplasma gondii, dirofilaria immitis, feline immunodeficiency virus (fiv), and feline leukemia virus (felv) infections among stray and pet cats in lanzhou, northwest china, and to identify the influence of age, gender, and regions on seropositivity. t. gondii antibodies were examined in cat sera by the modified agglutination test (mat). the circulating antigens of d. immitis and felv and specific antibodies to fiv were examined using kits ...201626362646
a case report of epstein-barr virus-associated retinal vasculitis: successful treatment using only acyclovir therapy.the purpose of this study was to describe a presumed case of epstein-barr virus (ebv)-associated retinal vasculitis in a 42-year-old female with sudden unilateral vision loss and successful treatment with acyclovir therapy. diagnostic vitreous biopsy of the right eye was performed to test for ebv and other known infectious causes of retinitis and evaluate vitreous cells and serological testing. vitreous polymerase chain reaction viral dna testing result was positive for ebv but negative for herp ...201627524923
schizophrenia and infections: the eyes have it.the visual tract is prominently involved in schizophrenia, as evidenced by perceptual distortions and a type of nystagmus found in many individuals affected. genetic explanations for these abnormalities have been suggested. this study proposes an alternate explanation based on infection. several infectious agents thought to be associated with some cases of schizophrenia are known to cause both infection of the fetus and abnormalities of the eye. toxoplasma gondii is examined in detail, and rubel ...201627507268
encephalitis caused by an unusual human herpes virus type 6 and toxoplasma gondii co-infection in a cord blood transplant recipient.the case of a central nervous system human herpes virus type 6 (hhv-6) and toxoplasma gondii co-infection after an umbilical cord blood transplantation in a chronic myelomonocytic leukaemia patient is reported.201627060193
latent toxoplasma infection and higher toxoplasma gondii immunoglobulin g levels are associated with worse neurocognitive functioning in hiv-infected adults.human immunodeficiency virus (hiv)-associated neurocognitive disorders persist despite suppressive antiretroviral therapy (art). because latent toxoplasma infection (lti) may adversely impact brain function, we investigated its impact on neurocognitive impairment (nci) in people living with hiv disease.201627794019
cerebral toxoplasmosis in patients with acquired immune deficiency syndrome in the neurological emergency department of a tertiary hospital.cerebral toxoplasmosis is the most common cause of space occupying brain lesion in patients with hiv/aids in brazil. in the post-haart era, it is responsible for high rates of morbidity and mortality worldwide.201627573702
continuous effector cd8(+) t cell production in a controlled persistent infection is sustained by a proliferative intermediate population.highly functional cd8(+) effector t (teff) cells can persist in large numbers during controlled persistent infections, as exemplified by rare hiv-infected individuals who control the virus. here we examined the cellular mechanisms that maintain ongoing t effector responses using a mouse model for persistent toxoplasma gondii infection. in mice expressing the protective mhc-i molecule, h-2l(d), a dominant t effector response against a single parasite antigen was maintained without a contraction p ...201627421704
toxoplasmosis and toxocariasis: an assessment of human immunodeficiency virus comorbidity and health-care costs in canada.toxoplasma gondii and toxocara spp. are zoonotic parasites with potentially severe long-term consequences for those infected. we estimated incidence and investigated distribution, risk factors, and costs associated with these parasites by examining hospital discharge abstracts submitted to the canadian institute for health information (2002-2011). annual incidence of serious toxoplasmosis and toxocariasis was 0.257 (95% confidence interval [ci]: 0.254-0.260) and 0.010 (95% ci: 0.007-0.014) cases ...201627139453
hepatitis c virus antibody titers associated with cognitive dysfunction in an asymptomatic community-based sample.hepatitis c virus (hcv) infection is associated with cognitive dysfunction in clinic-based studies. the risk could be attributed to factors such as antiviral medications, substance abuse, or coincidental infection.201627269819
elevated blood lead and cadmium levels associated with chronic infections among non-smokers in a cross-sectional analysis of nhanes data.experimental animal studies, in vitro experiments, and clinical assessments have shown that metal toxicity can impair immune responses. we analyzed data from a united states representative national health and nutrition examination survey (nhanes) to explore associations between chronic infections and elevated blood concentrations of lead and cadmium among non-smoking nhanes participants.201626864738
molecular investigation on the occurrence of toxoplasma gondii oocysts in cat feces using tox-element and its-1 region targets.one of the most important routes of transmission for toxoplasma gondii infection is the ingestion of foods contaminated with cat feces containing sporulated oocysts. the diagnosis of t. gondii infection by fecal microscopy is complicated, as other similar coccidian oocysts are often present in the same fecal specimen. this study aimed to identify t. gondii oocysts in cat feces using a novel pcr technique. feline fecal specimens (n = 254) were screened for coccidian oocysts by light microscopy us ...201627325616
seasonality in the proportions of domestic cats shedding toxoplasma gondii or hammondia hammondi oocysts is associated with climatic factors.a previous study on domestic cats in germany and neighbouring countries suggested seasonality in shedding toxoplasma gondii oocysts. the aim of the present study was to elucidate whether this seasonality in shedding could be explained by climatic effects and whether differences between years in the proportions of cats shedding oocysts could also be explained by climatic factors. to this end, a long-term study over a period of 55 months on domestic cats for t. gondii and hammondia hammondi oocyst ...201626820303
simultaneous detection of the protozoan parasites toxoplasma, cryptosporidium and giardia in food matrices and their persistence on basil leaves.toxoplasma gondii, cryptosporidium spp. and giardia intestinalis are emerging pathogen parasites in the food domain. however, without standardized methods for their detection in food matrices, parasitic foodborne outbreaks remain neglected. in this study, a new immunomagnetic separation assay (ims toxo) targeting the oocyst's wall of t. gondii was developed using a specific purified monoclonal antibody. performance of this ims toxo coupled to microscopic and qpcr analyses was evaluated in terms ...201627052700
toxoplasma gondii and giardia duodenalis infections in domestic dogs in new york city public parks.toxoplasma gondii and giardia duodenalis have been widely reported to produce major diseases in humans and domestic animals. little is known about the occurrence of these protozoan parasites in domestic dogs in the united states. the goal of this study was to determine the prevalence and genotypes of t. gondii and g. duodenalis among dogs in new york city. fecal samples from domestic dogs were analyzed by polymerase chain reaction (pcr). overall, 3% of the samples tested positive for t. gondii, ...201626988633
pathogen exposure varies widely among sympatric populations of wild and domestic felids across the united states.understanding how landscape, host, and pathogen traits contribute to disease exposure requires systematic evaluations of pathogens within and among host species and geographic regions. the relative importance of these attributes is critical for management of wildlife and mitigating domestic animal and human disease, particularly given rapid ecological changes, such as urbanization. we screened > 1000 samples from sympatric populations of puma (puma concolor), bobcat (lynx rufus), and domestic ca ...201627209780
myocarditis caused by feline immunodeficiency virus in five cats with hypertrophic cardiomyopathy.viral infections have been implicated as the cause of cardiomyopathy in several mammalian species. this study describes hypertrophic cardiomyopathy (hcm) and myocarditis associated with feline immunodeficiency virus (fiv) infection in five cats aged between 1 and 4 years. clinical manifestations included dyspnoea in four animals, one of which also exhibited restlessness. one animal showed only lethargy, anorexia and vomiting. necropsy examination revealed marked cardiomegaly, marked left ventric ...201626797583
causes of endogenous uveitis in cats presented to referral clinics in north carolina.to investigate the causes of endogenous uveitis in cats presenting to referral ophthalmology clinics in north carolina.201626559384
toxoplasma gondii seroprevalence: 30-year trend in an hiv-infected us military cohort.to determine if toxoplasma gondii igg antibody prevalence is declining in hiv-infected persons, we analyzed data (1984-2013) from the us military hiv natural history study. we found that t. gondii seroprevalence at enrollment was associated with age and decreased significantly after 1995 (p=0.004), similar to the general us population.201626499204
microsphere-based antibody assays for human parvovirus b19v, cmv and t. gondii.human parvovirus b19 (b19v), cytomegalovirus (cmv) and toxoplasma gondii (t. gondii) may cause intrauterine infections with potentially severe consequences to the fetus. current serodiagnosis of these infections is based on detection of antibodies most often by eia and individually for each pathogen. we developed singleplex and multiplex microsphere-based suspension immuno assays (sias) for the simultaneous detection of igg antibodies against b19v, cmv and t. gondii.201626746194
genetic characterization of toxoplasma gondii from autopsy proven cases of aids associated cerebral toxoplasmosis in south india.toxoplasma gondii (t.gondii) infection can be devastating in the immunodeficient causing high morbidity and mortality. due to limited availability of both diagnostic facilities and highly active antiretroviral therapy (haart), toxoplasmosis continues to be a significant problem amongst acquired immuno deficiency syndrome (aids) patients in india. while scanty literature is available on t. gondii isolates in animals in india, little is known about the genetic diversity of the parasite in humans. ...201626802459
seroepidemiological study of canine leishmania infantum and toxoplasma gondii infections in shanghai, china, and analysis of risk factors.the aim of this study was to determine the seroprevalence of leishmania infantum and toxoplasma gondii among household dogs in shanghai (the most important industrial and commercial city in china), and to assess the possible risk factors associated with the infection. during 2014-2015, a total of 408 sera were collected from healthy household dogs and tested for l. infantum and t. gondii infection using commercial elisa kits. the endemic characteristics according to gender, age group and breed w ...201627660861
sybr green-based detection of leishmania infantum dna using peripheral blood samples.parasitological methods for the diagnosis of visceral leishmaniasis (vl) require invasive sampling procedures. the aim of this study was to detect leishmania infantum (l. infantum) dna by real time-pcr method in peripheral blood of symptomatic vl patient and compared its performance with nested pcr, an established molecular method with very high diagnostic indices. 47 parasitologically confirmed vl patients diagnosed by direct agglutination test (dat > 3200), bone marrow aspiration and presented ...201627065603
targeting the cytochrome bc1 complex of leishmania parasites for discovery of novel drugs.endochin-like quinolones (elqs) are potent and specific inhibitors of cytochrome bc1 from plasmodium falciparum and toxoplasma gondii and show promise for novel antiparasitic drug development. to determine whether the mitochondrial electron transport chain of leishmania parasites could be targeted similarly for drug development, we investigated the activity of 134 structurally diverse elqs. a cohort of elqs was selectively toxic to amastigotes of leishmania mexicana and l. donovani, with 50% inh ...201627297476
protozoal infections of the cornea and conjunctiva in dogs associated with chronic ocular surface disease and topical immunosuppression.to describe five cases of protozoal keratitis or conjunctivitis in dogs with chronic preexisting ocular surface disease treated with long-term immunosuppression.201626037023
retail deli slicer cleaning frequency--six selected sites, united states, 2012.listeria monocytogenes (listeria) causes the third highest number of foodborne illness deaths (an estimated 255) in the united states annually, after nontyphoidal salmonella species and toxoplasma gondii (1). deli meats are a major source of listeriosis illnesses, and meats sliced and packaged at retail delis are the major source of listeriosis illnesses attributed to deli meat (4). mechanical slicers pose cross-contamination risks in delis and are an important source of listeria cross-contamina ...201627031689
meningoencephalitis and listeria monocytogenes, toxoplasma gondii and brucella spp. coinfection in a dolphin in italy.listeria monocytogenes, toxoplasma gondii and brucella spp. can infect a wide range of species, including humans. in cetaceans, meningoencephalitis has been associated with t. gondii and brucella spp. infection, whereas to our knowledge, l. monocytogenes infection has not previously been reported. meningoencephalitis and l. monocytogenes, t. gondii and brucella spp. were identified by means of both direct and indirect laboratory techniques in an adult female striped dolphin stenella coeruleoalba ...201626912047
killer lymphocytes use granulysin, perforin and granzymes to kill intracellular parasites.protozoan infections are a serious global health problem. natural killer (nk) cells and cytolytic t lymphocytes (ctls) eliminate pathogen-infected cells by releasing cytolytic granule contents--granzyme (gzm) proteases and the pore-forming perforin (pfn)--into the infected cell. however, these cytotoxic molecules do not kill intracellular parasites. cd8(+) ctls protect against parasite infections in mice primarily by secreting interferon (ifn)-γ. however, human, but not rodent, cytotoxic granule ...201626752517
seroprevalence of toxoplasma gondii and neospora caninum in red deer from central italy.neospora caninum and toxoplasma gondii are cosmopolite protozoan parasites impacting on human and animal health. in particular, t. gondii commonly infects human beings and all warm-blooded animals, while n. caninum is responsible for bovine abortion and neuromuscular disease in dogs. the aim of the presented survey was to evaluate the occurrence and prevalence of these parasites in the most numerous italian red deer population. the sera of 60 red deer (cervus elaphus) inhabiting central italy (4 ...201628030947
seroprevalence of toxoplasma gondii and neospora caninum in goats from north-western spain.toxoplasma gondii and neospora caninum are protozoans involved in reproductive failure especially in ruminant livestock. the objective was to estimate the seroprevalence of both parasites in goats from north-western spain and to study the influence of some factors on seropositivity.201628030928
occurrence of neospora caninum and toxoplasma gondii antibodies in dogs from rural properties surrounding a biological reserve, espirito santo, brasil.the aim of this study was to determine the presence of antibodies to neospora caninum and toxoplasma gondii in serum samples of 187 dogs from 30 rural properties surrounding córrego do veado biological reserve, espírito santo state, brazil. the reserve is one of the last lowland atlantic forest remnants of the region, surrounded by agriculture farms and cattle pastures. the presence of igg antibodies was determined by the indirect fluorescent antibody test for t. gondii (cut-off 1:16) and n. can ...201627925068
serosurvey for leishmania spp., toxoplasma gondii, trypanosoma cruzi and neospora caninum in neighborhood dogs in curitiba-paraná, brazil.neighborhood dogs may act as reservoirs for several zoonotic protozoan infections, particularly in urban areas, thus constituting a potential public health threat. accordingly, the aim of the present study was to evaluate the exposure of neighborhood dogs to four protozoan pathogens in public areas with high levels of human movement in curitiba, southern brazil. blood samples from 26 neighborhood dogs were screened by means of the indirect immunofluorescent antibody test (ifat) for leishmania sp ...201627925057
the common zoonotic protozoal diseases causing abortion.toxoplasmosis, neosporosis, sarcosporidiosis (sarcocystosis) and trypanosomiasis are the common zoonotic protozoal diseases causing abortion which caused by single-celled protozoan parasites; toxoplasma gondii, neospora caninum, sarcocystis spp and trypanosoma evansi, respectively. toxoplasmosis is generally considered the most important disease that causing abortion of both pregnant women and different female animals throughout the world, about third of human being population had antibodies aga ...201627876900
prevalence of toxoplasma gondii and neospora caninum in red foxes in slovakia.sera or meat juices of 177 red foxes (vulpes vulpes) originated from the localities of a human-influenced landscape (group 1) and 126 foxes from the protected mountain region (group 2) of slovakia, collected during 2010-2014 were tested for the presence of antibodies against toxoplasma gondii using indirect elisa and neospora caninum by competitive elisa. the tissue and uncoagulated blood samples were examined for the presence of the parasite's dna. the total seropositivity to t. gondii was 62.7 ...201627787208
vaccination against parasites - status quo and the way forward.although vaccination against various pathogens is integral to health management of swine, vaccines against parasites have not yet been commercialized for the use in pigs. the incentive to develop and commercialize anti-parasitic vaccines in swine are twofold; on the one hand parasitic diseases which are economically important, such as ascarosis and neonatal coccidiosis, could be controlled in a sustainable manner; on the other hand, the transmission of zoonotic parasites, such as toxoplasma gond ...201628405456
ocular toxoplasmosis ii: clinical features, pathology and management.the term, ocular toxoplasmosis, refers to eye disease related to infection with the parasite, toxoplasma gondii. recurrent posterior uveitis is the typical form of this disease, characterized by unilateral, necrotizing retinitis with secondary choroiditis, occurring adjacent to a pigmented retinochoroidal scar and associated with retinal vasculitis and vitritis. multiple atypical presentations are also described, and severe inflammation is observed in immunocompromised patients. histopathologica ...201622712598
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