Molecular Parasitology

A detailed discussion is presented highlighting the most common characteristic of the reptilians and their hybrid cohorts. It is elucidated that the negative aliens are highly, violent, warring, separatist, disconnected, narcissistic, selfish, egotistical, psychopathic and sociopathic form of beings with strong proclivity to anger and apathy for others who are utterly void of compassion, empathy, or love or emotions for that matter except anger. They also highly venerate physical strength above anything else and use it effectively to wage wars against others, conquer them and putting them in bondage for free labor. Being energetically parasitic-vampiric, not being able to sustain themselves against inevitable self-consumption, they use the same motivation and disposition with utter cunningness and deceptiveness in search of the conquest of solar organic sentient consciousness beings in order to siphon, harvest, and prey upon their energies as a food source in which they feel they need to keep them in a state of unconsciousness enslavement so they do not know what is happening to them. Being void of solar organic consciousness and unable to consciously vibrate, they are often very afraid of solar light or sunshine. They are soulless and void of any organic consciousness embodiment and are non-sentient beings unable to 'sense' the progression of time whatsoever. Thus, evolution is not a factor in their question of artificial existence nor is multidimensionality living multiple lives at the same time, being robots, cyborgs, automatons, and a form of dead energy. They exist as automatons surviving within a hive organization or society and they strictly adhere to a pyramidal hierarchical structure in which those deemed as stronger and superiors reside on progressively the higher pyramidal scale. They are also a highly envious, greedy, avaricious group of beings who perceive themselves as superior to others and entitled to pillage and plunder others' belongings and energetic resources including their consciousness energies upon which they are dependent to survive. This has made them essentially a glutinous lustful group of beings also prone to sloth, reaping the benefit of hard labor of the others. Also being such narcissists, self-supremacists, and elitists, they have digressed to believe in racial and other forms of prejudices including the repugnant dogma of eugenics. Finally, a most distasteful trait that they are associated with is that of delusional sense of male-supremacy giving rise to a patriarchal domination dogma of existence as well as misogyny, viewing the female as weak and "worthless" unlike their males as the epitome of physical strength, forever in search of hunting-gathering, war and conquest, as a means for distinction and veneration.

In the present study, the gastroprotective mechanism of Xylocarpus granatum fruit and its active constituents gedunin and photogedunin was investigated. Chloroform fraction (Fr-CHCl 3 ) of X. granatum fruit was evaluated against cold restraint (CRU), aspirin (AS), alcohol (AL) and pyloric ligation (PL) induced gastric ulcer models in rats and histamine (HA) induced duodenal ulcer model in guinea pigs. Potential anti-ulcer activity of Fr-CHCl 3 was observed against CRU (58.28%), AS (67.81%), AL (84.38%), PL (65.66%) and HA (61.93%) induced ulcer models. The standard drug omeprazole (10 mg/kg, p.o.) showed 68.25% protection against CRU, 57.08% against AS and 69.42% against PL model and 70.79% against HA induced duodenal ulcer. Sucralfate, another standard drug (500 mg/kg, p.o.) showed 62.72% protection in AL induced ulcer model. Fr-CHCl 3 significantly reduced free acidity (51.42%), total acidity (30.76%) and upregulated mucin secretion by 58.37% respectively. Phytochemical investigations of Fr-CHCl 3 yielded gedunin (36%), photogedunin (2%). Further, Fr-CHCl 3 and its compounds gedunin and photogedunin significantly inhibited H + K + -ATPase activity in vitro with IC 50 of 89.37, 56.86 and 66.54 mg/ml respectively as compared to the IC 50 value of omeprazole (30.24 mg/ml) confirming their anti-secretory activity. Conclusively, Fr-CHCl 3 of Xylocarpus granatum was found to possess anti-ulcerogenic activity which might be due to its anti-secretory activity and subsequent strengthening of the defensive mechanism. This study is the first of its kind to show significant anti-secretory effect of gedunin and photogedunin. Therefore it could act as a potent therapeutic agent against peptic ulcer disease.

Phenylhydrazine (PHZ), a potent chemical causes toxicity on various tissues at various levels. Administration of phenylhydrazine mainly causes haematotoxicity which leads to the haemolytic anemia. In mammals PHZ induced anemia increased the iron absorption in spleen, liver and duodenum and finally iron metabolism was altered. Local demand and supply of Fe would increase erythropoeitic activity of the spleen so the size of spleen was increased that create the splenomegaly. PHZ induced anemia activate immune response which triggers phagocytosis in the spleen and liver. Apart from this administration of PHZ interfere the binding of erythropoietin (EPO) with erythropoietin receptors (EPOR) so that JAK-STAT would be affected.PHZ also showed genotoxic effect by creating single strand DNA damage.

A B S T R A C T Therapeutic antibodies are widely used in the treatment of various diseases and disease conditions, including cardiovascular diseases, autoimmune disorders, malignancies, and infections. With at least 23 therapeutic agents currently in clinical use and a successful business generating large revenues, major technological advances are now in place to improve the specificity and efficacy of those antibodies already in the market and also generate new, safe and effective macromolecules for the treatment of other ailments. This review provides a summary of the current state of antibody therapy, highlights and discusses recent developments in the field of antibody-based therapeutics production, combination therapy and shows the status of some of the agents that are in clinical trial. ᭧

La  criptosporidiosis  es  una  infección  intestinal  en  humanos  causada  por  el parásito Cryptosporidium. Esta infección se presenta en todo el mundo, pero  los  más  susceptibles  son  quienes  tienen  un  sistema  inmune  débil,  como las personas con sida. La enfermedad se transmite a través de agua o alimentos contaminados con heces. Hasta el momento no hay un trata-miento efectivo contra esta parasitosis.

Hepatozoon parasites were examined for the first time in reptiles from the Seychelles Islands. Although both prevalence
and intensity were low, Hepatozoon species were detected in individuals from 2 endemic species, the lizard Mabuya wrightii and the snake Lycognathophis seychellensis. This was confirmed using visual identification and through sequencing part of the 18s rRNA gene. Phylogenetic analysis indicates that the Hepatozoon on the Seychelles form a monophyletic lineage, although more data are clearly needed to stabilize estimates of relationships based on this marker.

DNA metabarcoding offers new perspectives, especially with regard to the high‐throughput identification and diagnostics of pathogens. Microsporidia are an example of widely distributed, opportunistic and pathogenic microorganisms in which molecular identification is important for both environmental research and clinical diagnostics. We have developed a method for parallel detection of both microsporidian infection and the host species. We designed new primer sets: one specific for the classical Microsporidia (targeting the hypervariable V5 region of small subunit [ssu] rDNA), and a second one targeting a shortened fragment of the COI gene (standard metazoan DNA‐barcode); both markers are well suited for next generation sequencing. Analysis of the ssu rDNA data set representing 607 microsporidian species (120 genera) indicated that the V5 region enables identification of >98% species in the data set (596/607). To test the method, we used microsporidians that infect mosquitoes in natural populations. Using mini‐COI data, all field‐collected mosquitoes were unambiguously assigned to seven species; among them almost 60% of specimens were positive for at least 11 different microsporidian species, including a new microsporidian ssu rDNA sequence (Microsporidium sp. PL01). Phylogenetic analysis showed that this species belongs to one of the two main clades in the Terresporidia. We found a high rate of microsporidian co‐infections (9.4%). The numbers of sequence reads for the operational taxonomic units suggest that the occurrence of Nosema spp. in co‐infections could benefit them; however, this observation should be retested using a more intensive host sampling. Our results show that DNA barcoding is a rapid and cost‐effective method for deciphering sample diversity in greater resolution, including the hidden biodiversity that may be overlooked using classical methodology.

This study presents new findings concerning the evolution of the human pathogens, Trypanosoma brucei and T. cruzi, which suggest that these parasites have divergent origins and fundamentally different patterns of evolution. Phylogenetic analysis of 18S rRNA sequences places T. brucei in a clade comprising exclusively mammalian trypanosomes of African origin, suggesting an evolutionary history confined to Africa. T. cruzi (from humans and sylvatic mammals) clusters with trypanosomes specific to Old and New World bats, T. rangeli and a trypanosome species isolated from an Australian kangaroo. The origins of parasites within this clade, other than some of those from bats, lie in South America and Australia suggesting an ancient southern super-continent origin for T. cruzi, possibly in marsupials ; the only trypanosomes from this clade to have spread to the Old World are those infecting bats, doubtless by virtue of the mobility of their hosts. Viewed in the context of palaeogeographical evidence, the results date the divergence of T. brucei and T. cruzi to the mid-Cretaceous, around 100 million years before present, following the separation of Africa, South America and Euramerica. The inclusion in this study of a broad range of trypanosome species from various different hosts has allowed long phylogenetic branches to be resolved, overcoming the limitations of many previous studies. Moreover, T. brucei and the other mammalian tsetse-transmitted trypanosomes appear, from these data, to be evolving several times faster than T. cruzi and its relatives.

Gene imprinting has conduited the scope of our understanding of phenotypic expression and its corelation with constituent genotype. It is an epigenetic process that involves DNA methylation and histone modulation to attain monoallelic gene expression without altering the genetic sequences. A distinctive model of non-mendelian genetics, imprinting extends the control over expression of traits and selection of the allele that would direct the same, in a manner decided by the parent of origin. The constitutive existence of this imprinting even after gametogenesis, throughout the somatic development extends a clue for its regulatory hold on several heridetary traits. Several heridetary diseases like Cancers, Russell-Silver syndrome, Beckwith-Wiedemann syndrome, Prader-Willi and Angelman Syndromes and Neurodegenration have shown to be a subsequent error in the genomic impriting process. So, understanding these epigenetic regulations can be a therapeutic strategy for disease modelling and especially targeting their patterns of heridetary inheritance.

Many methods have been proposed for collecting and fixing parasites, but most were written before the molecular age, and were intended to be practised by experienced parasitologists in well-equipped laboratories. We describe here a very simple method, illustrated by photographs, for collecting helminths from the digestive tract of vertebrates. It only requires a few plastic vials, some ethanol and a means to heat water. Basically, the method consists of: (a) the extraction of all organs from the abdominal cavity; (b) opening the digestive system longitudinally; (c) agitate gut and contents in a saline solution (i.e. ca. 9% NaCl or ¼ sea water in tap water); (d) decant in saline as many times as needed to clean contents; (e) immediately fix parasites in near-boiling saline; (f) discard saline and keep specimens in 95% ethanol. Additional information is given for collecting parasites from fish gills with a similar process. The method will collect most helminths (digeneans, larval cestodes, nematodes, acanthocephalans) from the digestive tract, and monogeneans and isopod and copepod crustaceans from fish gills. The specimens will be suitable for both morphological study and DNA sequencing. The method is simple, fast, inexpensive, and can be used by untrained personnel, even in the field without electricity and without a binocular microscope. It can also be used by trained parasitologists who need to expedite treatment of abundant samples.

Diarrhea is the most common child disease; one of the most prominent causes is Giardiasis, which produces an infection of intestinal characterized by abdominal pain, bloating, nausea, and episodes of diarrhea. Giardiasis is produced by unicellular parasites called Giardia lambila that can be found all over the world, especially in areas of poor sanitation, unsafe water, the current study aimed at investigating the infection of Giardia lambila in school children with diarrhea and who were referred by their relatives to the women's hospital and children in Maysan governorate south of Iraq, (200) samples of feces were examined for infants patientswith diarrhea in(0-2) years,the microscopic examination of this samples showed positive infection with Giardia lambila 80/200 (40%), the study of biochemical and blood parameters of the patients showed a decrease in the concentration of hemoglobin level, packed cell volume and the increase in the differential number of WBC in the infected infants compared to the control group, the biochemical study also indicated a decrease in the concentration of total protein, albumin, calcium, magnesium and sodium ions, cholesterol, triglycerides, and increase in concentration of liver enzymes GTP and GOT in patients compared to the control group.

Biological membranes define the boundaries of cells and compartmentalize the chemical and physical processes required for life. Many biological processes are carried out by proteins embedded in or associated with such membranes. Determination of membrane protein (MP) structures at atomic or near-atomic resolution plays a vital role in elucidating their structural and functional impact in biology. This endeavor has determined 1198 unique MP structures as of early 2021. The value of these structures is expanded greatly by deposition of their three-dimensional (3D) coordinates into the Protein Data Bank (PDB) after the first atomic MP structure was elucidated in 1985. Since then, free access to MP structures facilitates broader and deeper understanding of MPs, which provides crucial new insights into their biological functions. Here we highlight the structural and functional biology of representative MPs and landmarks in the evolution of new technologies, with insights into key developments influenced by the PDB in magnifying their impact. As membranes were critical to the separation of chemistries essential to the origin of life, membrane proteins (MPs) are key players in some of the most important physiological processes in living organisms. Characterizing MPs structurally and functionally is still extremely challenging due to their frequent low abundance, and difficulties in purifying functional MPs intact from their native membrane, though it is going through an exciting revolution now due to several key factors. It took several decades to obtain the structural information that now allows pursuit of understanding MP function in health and disease, to manipulate them as drug targets, and to engineer them into new powerful tools to fuel discovery. We highlight some of the landmarks in this endeavor that drove or depended on the discovery of new technologies required specifically for structural studies of membrane, versus soluble proteins. Previously, handwritten letters delivered by post requested coordinate sets that were not always readily given. The vision of Hamilton, Myer, Koetzle, and the joint venture between the Cambridge Crystallographic Data Center in the United Kingdom and the Brookhaven National Laboratories at Stony Brook University led to the Protein Data Bank (PDB). Then for the first time, one could begin to ask questions with all the relevant structures at hand. In celebrating 50 years of the PDB, we focus on some of the ingeniously crafted inventions and discoveries that led the way for entire classes of MPs, and those new approaches that now promise structures of large and complex machines from their native cellular environments, in action. In the following Historical perspective, we provide a brief historical perspective of some MP insights (other than the crucial G-protein-coupled receptors (GPCRs) that are the subject of a dedicated review in this volume) and consider the value of the PDB in disseminating this information. Seemingly insurmountable difficulties were often overcome with invention of new technologies to reveal important structural features of classes of MPs that make the fabric of today's approaches. In How do membrane proteins accomplish key physiological functions?, we describe how the structures of several of the major MP classes were uncovered, which often required technological developments that are now woven into the fabric of structural biology. First, how are the α-helical, tail-anchored, and all β-sheet MP broad categories correctly targeted to and inserted into membranes and allowed to fold correctly? We progress to landmark discoveries involving the roles of MPs in physiology and some of the critical barriers that had to be overcome to realize these achievements. How do water channels conduct water at diffusion limited rates without allowing leakage of protons (H +) or hydronium ions (H 3 O +) or any other ions? How do potassium channels conduct K + ions, but ‡ These authors contributed equally.

During a survey from June 2011 to May 2012, 60 Catla catla (Hamilton-Buchanan) were collected from Harike wetland of Punjab, out of which gills of 35 fishes were found to be infected with myxobolid parasites belonging to the genus: Myxobolus (Myxozoa: Myxosporea: Myxobolidae). Total infection rate was found to be 58.3%. In total, six myxobolid species were found infecting gills of Catla catla. Maximum prevalence was seen in the month of May (100%) followed by October (80%), June (69.23%), July (60%), August and April (66.6%) and July (60%); minimum in December and February (33.3%). Maximum mean intensity and abundance have been shown in the month of October 39.5% & 31.6% respectively, on the other hand least intensity and abundance was shown in the month of August 12% and 8% respectively. The maximum relative prevalence and overall prevalence of Myxobolus naini Kaur and Singh (2008) was 42.85% and 25% respectively and minimum in M. saranae Gupta and Khera (1990) and M. filamentosus (Haldar et al., 1981) Gupta and Khera, 1988a i.e. 5.7% and 3.3% was recorded.

PCR-RFLP techniques have been used to characterize wild and laboratory isolates of the trichostrongyle nematode Heligmosomoides polygyrus from the wood mouse Apodemus sylvaticus and the laboratory mouse Mus musculus respectively. Both isolates can be distinguished by eight endonuclease digestions of the ITS region of the rDNA repeat namely, Alu I, Dde I, Hpa II, Hae III, Hinf I, Hha I, Pvu II and Sal I. In two of the digests, Hinf I and Rsa I, a minor polymorphism was observed in the wild isolate of H. polygyrus which has been cultured in laboratory-bred A. sylvaticus for several generations when compared with H. p. polygyrus from wild A. sylvaticus. A minor polymorphism was also identified in further wild isolates of H. polygyrus collected from A. sylvaticus in a field site in Egham, Surrey. However no evidence of polymorphism was observed in the laboratory isolate of H. polygyrus from the CD1 strain of M. musculus and the laboratory-bred A. sylvaticus. Reasons for this are discussed and further studies on the population genetics of H. polygyrus are suggested.

Background: A wide range of molecules are used by tapeworm metacestodes to establish successful infection in the hostile environment of the host. Reports indicating the proteins in the cestode-host interactions are limited predominantly to taeniids, with no previous data available for non-taeniid species. A non-taeniid, Hymenolepis diminuta, represents one of the most important model species in cestode biology and exhibits an exceptional developmental plasticity in its life-cycle, which involves two phylogenetically distant hosts, arthropod and vertebrate. Results: We identified H. diminuta cysticercoid proteins that were recognized by sera of H. diminuta-infected rats using two-dimensional gel electrophoresis (2DE), 2D-immunoblotting, and LC-MS/MS mass spectrometry. Proteomic analysis of 42 antigenic spots revealed 70 proteins. The largest number belonged to structural proteins and to the heat-shock protein (HSP) family. These results show a number of the antigenic proteins of the cysticercoid stage, which were present already in the insect host prior to contact with the mammal host. These are the first parasite antigens that the mammal host encounters after the infection, therefore they may represent some of the molecules important in host-parasite interactions at the early stage of infection.

In the present study, a new species Myxobolus dermiscalis n. sp. infecting scales of Labeo rohita, an Indian major carp from Harike Wetland in Punjab, India has been described on the basis of spore morphology and amplification of a part of 18S rDNA gene. The pseudocysts of M. dermiscalis n. sp. are milky white with irregular outline, 0.5e3.6 mm in diameter embedded within the dermal scale in the form of a cavity. The spores 5.84e7.98 Â 3.98e5.98 mm in size, having two equal polar capsules 3.98e5.98 Â 1.85 e3.85 mm in size. The most differentiating feature from closely related species, Myxobolus saugati (Kaur and Singh, 2011) is the presence of two parietal folds at the posterior e lateral margins of the shell valves. The present species is regarded as host, organ and tissue specific in nature. The partial sequence of SSU gene of M. dermiscalis n. sp. clustered with other Myxobolus species infecting cyprinids available in the GenBank. Blast search revealed 98% homogeneity with Myxobolus sp (KM401439) infecting scales of L. rohita in Myanmar (unpubl. data). The present myxobolid parasite has been recorded to cause serious, highly symptomatic disease of the scales, causing their loosening from the skin of L. rohita. It rendered the host fish unsightly giving it cloudy appearance with white patches and mucoid body surface. Scale pseudocyst Index (SPI) has been provided to record the intensity of infection.

Background: Trypanosoma brucei is the causative agent of human sleeping sickness and animal trypanosomiasis in sub-Saharan Africa, and it has been subdivided into three subspecies: Trypanosoma brucei gambiense and Trypanosoma brucei rhodesiense, which cause sleeping sickness in humans, and the nonhuman infective Trypanosoma brucei brucei. T. b. gambiense is the most clinically relevant subspecies, being responsible for more than 90% of all trypanosomal disease in humans. The genome sequence is now available, and a Mendelian genetic system has been demonstrated in T. brucei, facilitating genetic analysis in this diploid protozoan parasite. As an essential step toward identifying loci that determine important traits in the human-infective subspecies, we report the construction of a high-resolution genetic map of the STIB 386 strain of T. b. gambiense.

The prevalence of bovine cysticercosis was established using routine postmortem inspection of 3450
carcasses of buffaloes slaughtered in 2014 in Kaliouba governorate, among which 313 (9.07%) were detected
as harbouring cysticercosis lesions using meat inspection process. The cysts were examined macroscopically
for description of their morphology and constituents and classified as viable or degenerating. Viable cysts were
microscopically confirmed for demonstration of protoscolex. Out of 100 of patients offered taenicidal drugs
examined by microscopic examination through direct and sedimentation of fecal samples, 6 (6%) were positive
for Taenia saginata (T. saginata) eggs. Histological sections of 6 gravid proglottids were identified as
T. saginata. We used a biomolecular assay targeting the HDP2 gene for developing PCR assay in 20 viable
cysts and 6 gravid proglottids. An HDP2 gene-PCR amplification product of the taeniid samples of T. saginata
is approximately 599bp. Partial sequences were generated after gel purification of PCR amplified products of
HDP2 gene with sequence analysis and subsequent phylogeny to compare these sequences to those from
known strains of T. saginata circulating globally and retrieved from GenBank. Most isolates with accession No.
KT027580 are closely related to T. saginata based on the similarity of nucleotide sequences and phylogenetic
relationships. In conclusion, this work indicated high prevalence rate of bovine cysticercosis and T. saginata,
both morphological examination of the parasite and molecular analysis using bioinformatic tools identified the
metacestode and revealed typical taeniid features confirmed to Taenia saginata

Aim: This study was undertaken to identify Paramphistomum cervi on the basis of its morphology and histology to be
the common cause of paramphistomosis in infected sheep and its differentiation from other similar Paramphistomes in
Gujarat.
Materials and Methods: Adult rumen flukes were recovered from the rumen of naturally infected sheep slaughtered in
various abattoirs in Gujarat. Some adult flukes were flattened and stained in Borax carmine, and some were sectioned in the
median sagittal plane and histological slides of the flukes were prepared for detailed morphological and histological studies.
Result: Microscopic pictures of the parasite used in identification define the similarity in the morphology and histology of
the anterior sucker, pharynx, esophagus, genital atrium, posterior sucker (acetabulum) and testes to the P. cervi.
Conclusion: It can be concluded that the most common species found in sheep infected with Paramphistomosis is P. cervi
on the basis of its histo-morphological appearance in Gujarat.

Invitro sensitivity of two isolates of Entamoeba histolytica from New Delhi to antiamoebic drugs Metronidazole and Auranofin.
Metronidazole is a antiprotozoal agent of the nitroimidazole class, is used against protozoans such as Trichomonas vaginalis, Entamoeba, and Giardia. Auranofin, a organogold compound classified by the World Health Organization as an antirheumatic agent, was identified as active against the Entamoeba histolytica. Auranofin was 10-times more potent against Entamoeba histolytica than Metronidazole when tested in axenic cultures of entamoeba.
In this study, we tried to find out the tolerance and efficacy of these two drugs against clinical isolates of Entamoeba histolytica grown xenically.
Minimum Inhibitory Concentration of Metronidazole and Auranofin of drug were determined by Invitro sensitivity assay, using the microtiter plates. Cell scoring was given by microscopic observation under an inverted microscope. To confirm their MICs, cell were stained using trypan blue and the viable parasites where counted by using heamocytometer.
The results show that the isolates were able to tolerate metronidazole at high concentration however Auranofin proved to be the efficacy at low concentration against the clinical isolate of Entamoeba histolytica grown in Xenic cultures.

In humans and other eukaryotes, histone post-translational modifications (hPTMs) play an essential role in the epigenetic control of gene expression. In trypanosomatid parasites, conversely, gene regulation occurs mainly at the post-transcriptional level. However, our group has recently shown that hPTMs are abundant and varied in Trypanosoma cruzi, the etiological agent of Chagas Disease, signaling for possible conserved epigenetic functions. Here, we applied an optimized mass spectrometry-based proteomic workflow to provide a high-confidence comprehensive map of hPTMs, distributed in all canonical, variant and linker histones of T. cruzi. Our work expands the number of known T. cruzi hPTMs by almost 2-fold, representing the largest dataset of hPTMs available to any trypanosomatid to date, and can be used as a basis for functional studies on the dynamic regulation of chromatin by epigenetic mechanisms and the selection of candidates for the development of epigenetic drugs against tr...

The present study was performed to analyze molecularly the phylogenetic positions of human-infecting Trichostrongylus species in Mazandaran Province, Iran, which is an endemic area for trichostrongyliasis. DNA from 7 Trichostrongylus infected stool samples were extracted by using in-house (IH) method. PCR amplification of ITS2-rDNA region was performed, and products were sequenced. Phylogenetic analysis of the nucleotide sequence data was performed using MEGA 5.0 software. Six out of 7 isolates had high similarity with Trichostrongylus colubriformis, while the other one showed high homology with Trichostrongylus axei registered in GenBank reference sequences. Intra-specific variations within isolates of T. colubriformis and T. axei amounted to 0-1.8% and 0-0.6%, respectively. Trichostrongylus species obtained in the present study were in a cluster with the relevant reference sequences from previous studies. BLAST analysis indicated that there was 100% homology among all 6 ITS2 sequences of T. colubriformis in the present study and most previously registered sequences of T. colubriformis from human, sheep, and goat isolates from Iran and also human isolates from Laos, Thailand, and France. The ITS2 sequence of T. axei exhibited 99.4% homology with the human isolate of T. axei from Thailand, sheep isolates from New Zealand and Iran, and cattle isolate from USA.

The Enzyme-Linked Immunosorbent Assay (ELISA) was evaluated for the diagnosis of Fasciola gigantica infection in cattle and buffaloes. The excretory-secretory (E-S Ag) antigen of Fasciola gigantica adult flukes obtained after invitro incubation was used as an antigen. The test was conducted with 276 sera collected from cattle and buffaloes which included 22 sera each from naturally infected cattle and buffaloes (known positive serum) and with similar number of samples with healthy cattle and buffaloes (known negative serum). The positive results were observed in 18 and 19 of the sera from naturally infected cattle and buffaloes with sensitivity of 81.8% and 86.3% respectively. Out of 188 serum samples which were found negative on faecal examination 32 (34%) sera of cattle and 40 (42.5%) sera of buffaloes were found positive by ELISA respectively. The sensitivity of the test was found to be 91.6% and 95.6% in cattle and buffaloes respectively.

The development of molecular methods is becoming a promising field in the analysis of parasite fauna in wildlife species. This is especially useful in the case of parasite species where developmental larval stages are difficult to assess using standard methods. In this study we screened for the presence of parasitic nematodes infecting lacertid lizards from the Iberian Peninsula and Balearic islands using nematode-specific 18S ribosomal RNA gene primers on host tissue samples. Sequencing of positive samples revealed the presence of different genera of nematodes. The detection of Strongyloides, a monoxenous genus reported for the first time in Podarcis lilfordi is probably the result of the amplification of larval stages present in the host circulatory system. Two spirurid nematodes, Synhimantus and a new unidentified clade, were also found, suggesting that reptiles might be paratenic hosts of several spirurid species. This study demonstrates the benefits of using specific molecular markers on tissue samples to identify infecting larval stages of nematodes, otherwise difficult to assess using traditional screening methods.(Received July 15 2011)(Accepted February 23 2012)(Online publication April 02 2012)

order with a spectrum of symptoms caused by primary cilium dysfunction. The disease is caused by mutations in one of at least 17 identified genes, of which seven encode subunits of the BBSome, a protein complex required for specific trafficking events to and from the primary cilium. The molecular mechanisms associated with BBSome function remain to be fully elucidated. Here, we generated null and complemented mutants of the BBSome subunit BBS1 in the protozoan parasite, Leishmania. In the absence of BBS1, extracellular parasites have no apparent defects in growth, flagellum assembly, motility or differentiation in vitro but there is accumulation of vacuole-like structures close to the flagellar pocket. Infectivity of these parasites for macrophages in vitro is reduced compared with wild-type controls but the null parasites retain the ability to differentiate to the intracellular amastigote stage. However, infectivity of BBS1 null parasites is severely compromised in a BALB/c mouse footpad model. We hypothesize that the absence of BBS1 in Leishmania leads to defects in specific trafficking events that affect parasite persistence in the host. This is the first report of an association between the BBSome complex and pathogen infectivity.

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Free pre-tetrathyridia of Mesocestoides sp. are described, for the first time, from samples obtained from the coelomic cavity of a ground skink, Scincella lateralis, from Oklahoma. Closer examination of these early-stage tapeworms revealed they were transitional metamorphosis stages between a post-hexacanth procercoid form to the full metacestode of Mesocestoides. A series of transitional stages was found that span the full period of sucker and apical organ development. However, we did not see any fully developed tetrathyridia, i.e., having classic Mesocestoides morphology but with the apical sucker absent following developmental atrophy. This is the first time that metamorphic pre-tetrathyridial stages of a Mesocestoides sp. have been reported in vivo from a natural infection. These observations corroborate earlier reports of such stages of Mesocestoides vogae developed in vitro, though the previously reported isolate of M. vogae is asexually proliferative, and the species from the present study showed no sign of asexual proliferation. The fact that these immediately post-hexacanth stages can occur in a single lizard intermediate host may suggest that Mesocestoides spp. might develop through a simple 2-host life cycle rather than an obligate 3-host cycle that has been speculated to occur by most previous authors. DNA sequence comparisons and phylogenetic analyses based on mitochondrial 12S, cox1, and nad1 genes have demonstrated that our specimens from S. lateralis represent a species clearly distinct from all previously sequenced Mesocestoides and closely related to 2 forms from domestic dogs and Channel Island fox in California previously published as Mesocestoides sp. C.

Background: Myxozoa is a well-known economically and ecologically important group of metazoan parasites, phylogenetically related to Cnidaria. High diversity of myxosporeans has been recorded in Russia and China; however, most of the species were solely morphologically characterised. Here, we identified a new gibel carp-infecting Myxobolus species and morphologically and molecularly compared the Russian and Chinese isolates of this new myxosporean.

A member of the Sparidae family, the goldline sea bream Sarpa salpa (Linnaeus, 1758) collected from the Gulf of Tunis and the Bay of Bizerte in Northeast Tunisia, Western Mediterranean, were examined for the myxozoan parasites. During the parasitological exposure, a total of 7 myxosporean are found including 6 coelozoic species belong to the genus Ceratomyxa Thélohan, 1892 infected the gallbladder of their host of which 3 known species have been previously described, C. arcuata Thélohan, 1892, C. pallida Thélohan, 1895 and C. herouardi Georgévitch, 1916 and 3 species seem different in morphology to Ceratomyxa spp already known from Mediterranean Sea or from other localities in the wide world. These species are Ceratomyxa sp. 1, Ceratomyxa sp. 2 and Ceratomyxa sp. 3. Only one histozoic species belongs to the genus Henneguya Thélohan, 1892, Henneguya sp. identified for the first time infecting the mesentery vessels of S. salpa. The myxosporean parasite C. arcuata Thélohan, 1892 is reported for the first time in Tunisian waters from the goldline sea bream which represents as new host records. In addition to the Monoparasitism, the phenomenon of Polyparasitism was observed between the current species in both sampling sites with two types: Biparasitism and Triparasitism. The most frequent Polyparasitism was a Biparasitism-type between C. herouardi and C. pallida with frequency 16.97%. For all the species, no serious pathogenic changes have been recorded in the host organ or in the outward appearance of the fish. Morphological features, site of infection into the host, parasite prevalence and mean intensity of each myxosporean found during this survey are determined and their taxonomic affinities to other species are discussed.

Malaria merupakan penyakit yang dipengaruhi oleh empat faktor, yaitu parasit (agent), manusia (hospes), nyamuk Anopheles (vektor), dan lingkungan (environment).  Penyakit ini ditularkan melalui gigitan nyamuk Anopheles sp.  Banyaknya tambak terlantar pada pantai di Desa Sukamaju Kecamatan Punduh Pedada Kabupaten Pesawaran diduga merupakan tempat perindukan nyamuk Anopheles sp.  Tujuan penelitian  ini untuk  mengetahui  hubungan antara faktor abiotik dengan kepadatan larva dan kondisi ekologi tempat perindukan vektor (tambak terlantar).  Penelitian ini dilaksanakan pada bulan April – Mei 2012 dengan metode survai.  Pengamatan berupa beberapa faktor ekologi (fisik, kimia, dan biologi) di tempat perindukan nyamuk serta pengambilan larva nyamuk untuk dihitung kepadatannya dan diidentifikasi.  Data kepadatan larva dan faktor abiotik dianalisis menggunakan korelasi pearson.  Hasil penelitian menunjukkan bahwa faktor abiotik seperti suhu air, oksigen terlarut (DO), dan kedalaman air memiliki korelasi terhadap kepadatan larva Anopheles sp. pada taraf  nyata 5%.  Kondisi ekologi perindukan nyamuk mendukung kehidupan larva dengan suhu tertinggi 31,330C, salinitas 2,78 ‰, pH berkisar 5-6, DO berkisar 5- 6,23 mg/L, dan kedalaman air  tertinggi 57,83 cm.  Kepadatan  larva paling tinggi terdapat pada tambak 3 yaitu 15,3 ekor/250 ml.  Pengamatan faktor biologi ditemukan jenis tumbuhan air berupa ganggang dan lumut serta hewan air berupa kepiting (Uca pugnax), udang (Palaemonete sp), kecebong (Rana sp.), dan bentos.
Kata kunci : Ekologi, Perindukan Vektor Malaria, Punduh Pedada

The present study is aimed to evaluate antifilarial activity of Xylocarpus granatum (fruit from Andaman) against human lymphatic filarial parasite Brugia malayi in vivo. The in vitro antifilarial activity has already been reported earlier for this mangrove plant which has traditionally been used against several ailments. Aqueous ethanolic crude extract, four fractions (Ethyl acetate fraction, n-butanol fraction, water soluble fraction and water insoluble fraction) and pure molecule/s of Xylocarpus granatum (fruit) were tested in vitro on adult worms and microfilariae (mf) of B. malayi and the active samples were further evaluated in vivo in B. malayi i.p. transplanted jird model (Meriones unguiculatus) and Mastomys coucha subcutaneously infected with infective larvae (L3). The crude aqueous ethanolic extract was active in vitro (IC50: adult=15.46 μg/ml; mf=13.17 μg/ml) and demonstrated 52.8% and 62.7% adulticidal and embryostatic effect on B. malayi respectively in mastomys at a dose of 5x250 mg/kg by oral route. The antifilarial activity was primarily localized in the ethyl acetate soluble fraction which revealed IC50 of 8.5 and 6.9 μg/ml in adult and mf respectively. This fraction possessed moderate adulticidal and embryostatic action in vivo in mastomys. Out of eight pure molecules isolated from the active fraction, two compounds Gedunin (IC50=0.239, CC50=212.5, SI=889.1) and Photogedunin (IC50=0.213, CC50=262.3, SI=1231.4) at 5x100 mg/kg by subcutaneous route revealed excellent adulticidal efficacy resulting in to the death of 80 % and 70 % transplanted adult B. malayi in the peritoneal cavity of jirds respectively in addition to noticeable microfilaricidal action on the day of autopsy. The findings reveal that the extract from the fruit Xylocarpus granatum contains promising in vitro and in vivo antifilarial activity against human lymphatic filarial parasite B. malayi which could be attributed to the presence of two pure compounds Gedunin and Photogedunin.

The study aimed at examining the molluscicidal activity against biomphalaria pfeifferi response to ethanolic extracts of some medicinal plants (Arachis hypogaea and Anthocleista djalonensis). In the last two decades, plant molluscicides have received considerable attention in the search for cheaper, effective, environmentally-friendly alternatives to expensive, imported chemotherapeutic agents and synthetic molluscicides used in schistosomiasis control. The snails used for the study were collected at Ibiade and Efire water stream in Ibiade community at Ijebu water side area of Ogun state while the two plants were collected from a nearby market in Ijebu Ode Local Government area of Ogun state. The ethanol extracts of the two plants were subjected to phytochemical investigations to detect their chemical constituents. Data obtained were subjected to Analysis of variance (ANOVA) using Statistical Package for Social Science (SPSS) version 20.0 and means were separated according to Duncan's Multiple Range Test (DMRT) at < 0.05probability level. Results revealed that the applied plant extracts (Anthocleista djalonensis and Arachis hypogeal) showed high antischistosomal activity and Anthocleista djalonensis showed the highest percentage of mortality (100%) after showed that the methanolic extracts of Anthocleista djalonensis and Arachis hypogeal, had high contents of tannin, saponin, alkaloids, flavonoid, phenols phytate and oxalate. Sequel to the findings of the study, it was recommended that there should be adequate and more information on the use of plant molluscicides on the cost-effectiveness and chronic toxicity, and difficulties of developing viable snail control programmes in rural areas using local resources.

Hemolivia mauritanica is an apicomplexan parasite infecting the tortoises Testudo graeca and Testudo marginata as the intermediate vertebrate host, and the tick Hyalomma aegyptium as the definitive invertebrate host. Here we used molecular techniques to detect an apparent H. mauritanica in the ticks H. aegyptium attached to Testudo graeca from North Africa. Sequences from 18S rRNA were used to place H. mauritanica in a phylogenetic framework. A single Hemolivia haplotype was recovered from 16 ticks. Prevalence was estimated at 22.6% of hosts having ticks infected with H. mauritanica.

The genus Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina) is composed of intracellular haemogregarine parasites
that are widely distributed among all tetrapod groups. The present study combines microscopic and molecular data on haemogregarine parasites from lizards in the western Mediterranean. We screened tissue samples and examined blood smears for the presence of species of Hepatozoon from four lizards, namely Algyroides marchi Valverde, endemic to Southeast Spain, Podarcis bocagei Seoane from Spain and Portugal, P. hispanica Steindachner from Spain, and P. lilfordi Günther from Cabrera, Balearic Islands (Spain). Our results show that prevalence and intensity of Hepatozoon parasites vary between and within lizard species from different regions. Algyroides marchi and P. bocagei from Spain had the lowest values, whereas P. hispanica had the highest. Phylogeny based on 18S rRNA gene sequences indicates that most of the new Hepatozoon sequences are part of a clade exclusive from North African and Iberian lizards, except for a single P. bocagei isolate that is found related to another clade including isolates from other reptile host species and rodents. Interestingly, isolates from Algyroides form a distinct monophyletic subgroup, which could be a signal of strict host-specificity within this host genus.

In 2005, an epidemic of the cestode Triaenophorus crassus in Arctic charr, Salvelinus umbla from Lake Grundlsee (Austria) was reported, resulting in the closure of the local fisheries. The parasite uses a copepod and a salmonid fish as intermediate hosts, and the European pike (Esox lucius) as definitive host. In Lake Grundlsee procercoids of T. crassus, T. nodulosus and Eubothrium salvelini use the copepod Cyclops abyssorum praealpinus as their first intermediate host. We report on the development of a PCR-RFLP assay for the specific differentiation between the procercoids of the three cestode species. Ban I restriction sites within a 207-bp 18S rRNA-amplified fragment yielded two and three species-specific products to unequivocally distinguish between T. crassus and T. nodulosus, respectively. The assay is sufficiently sensitive to characterise a single procercoid of both Triaenophorus species in their copepod host, enabling a direct measure of infection. This assay will be useful in monitoring the progress of the control methods currently in place.

Trichomoniasis is a sexually transmitted disease afflicting many women worldwide. The disease is usually not life-threatening, but has been associated with the development many serious health problems. The development of resistant Trichomonas vaginalis strains necessitates the development of other nontraditional medications. Studies on deer musk showed its anti-bacterial and anti-fungal affects. This study was carried out to evaluate in vitro effects of different concentrations of musk 10, 15, 20, 25 and 30% (vol/vol) for 24, 48, 72and 96 hours at 37 °C on multiplication, viability and motility of Trichomonas vaginalis also morphological changes using scanning and transmission electron microscopy were reported and optical density of treated cultures were assayed. The minimal inhibitory concentrations of musk were 10% at 96h, 15% at 72h, 20% and 25% at 48h and 30% at 24h. The calibration curve of musk studied concentrations was found to be linear (R 2 = 0.942) indicating large positive association between the concentration of musk and the optical density of the culture.Musk is a safe natural product having the privilege of being anti Trichomonas vaginalis as well as antifungal and antibacterial properties so it provides a promising source for new drugs development.

The burden of hepatocellular carcinoma (HCC) in Egypt has been increasing with a doubling in the incidence rate in the past 10 years, which necessitates the investigation of the possible risk factors to its development. The present study aimed at investigating the role of Schistosoma mansoni infection as a risk factor for development of HCC. Five hundred parasite free mice were categorized into four groups: Group I (induction of carcinoma by diethylnitrosamine (DEN)), Group II (DEN + Infection), Group III (Infection) and Group IV (Control). Groups I and II were further subdivided into 4 subgroups according to the dose of DEN given. Serum samples from each group were examined for levels of tumor markers alpha fetoprotein (AFP) and ferritin by ELISA, then mice were sacrificed and subjected to histopathological examination of their livers. These were repeated every week till the end of the experiment. The results of the histopatho-logical examination clarified the role of S. mansoni in enhancing and aggravating the carcinogenic effect of DEN; dysplastic changes appeared earlier, with a higher grade and with a smaller dose of DEN in Group II compared to Group I. Serum levels of tumor markers showed earlier statistically significant differences in Group II than in Group I when compared to Group IV. We conclude that S. mansoni accelerates hepatic dysplastic changes in the presence of other risk factors making cancer appear early and with a more aggressive nature, compared to the same risk in absence of schistosomiasis.

Background: Malaria in Cameroon was previously known to be caused solely by Plasmodium falciparum but today, evidence points to other Plasmodium species including P. vivax, P. ovale and P. malariae. The purpose of this study was to identify the Plasmodium species in clinical samples from children residing in five epidemiological strata of malaria in Cameroon, so as to advise control policies.
Methods: One thousand six hundred nine febrile children (≤15 years) were recruited from five epidemiological strata of malaria including the Sudano-sahelian (SS) strata, the High inland plateau (HIP) strata, the South Cameroonian Equatorial forest (SCEF) strata, the High western plateau (HWP) strata and the Coastal (C) strata. Malaria parasites were detected by Giemsa microscopy (GM) while a multiplex polymerase chain reaction (PCR) was used to identify the Plasmodium species. Statistical analysis performed included the Pearson chi-square test, and statistical significance was set at p < 0.05.
Results: The PCR-adjusted prevalence of malaria was 17.6%. The detection rate of PCR was higher than GM (p = 0.05). However, GM demonstrated a high sensitivity (85.5%) and specificity (100%) and, overall, a perfectly correlated agreement with PCR (97.5%). The prevalence of malaria was significantly higher in children between 60 and 119 months (p < 0.001) and in Limbe (in the Coastal strata) (p < 0.001). Contrariwise, the prevalence of malaria was not associated with gender (p = 0.239). P. falciparum was identified in all (100%) the cases of malaria; P. ovale, P. vivax, P. malariae and P. knowlesi were all absent. No case of mixed infection was identified.
Conclusions: P. falciparum was the only species causing clinical malaria in the target population, which is contrary to studies that have reported P. vivax, P. malariae and P. ovale as causing clinical malaria in Cameroon.