Lymphocyte Migration Inhibition Response in Trichuris muris Infected and Vaccinated Mice

Background

Immunological response of host and parasite play a key role in developing vaccination and immunization. The present study deals with the immune response and effecter mechanism, which was confirmed by migration inhibition factor (MIF).

Methods

The present work was conducted in Parasitological Lab of Postgraduate Department of Zoology, Government Holkar Science College, Indore (M.P.) during 2006–2007. For MIF assay, lymphocytes were separated from heparinized blood of experimental and control mice. Aliquots of cell suspension were placed in four wells cut in a preparation of agarose in a Petri dish. Two wells were filled with soluble test antigen, while rest two wells were filled with medium (control wells). Petri dish was incubated overnight at 37 °C in a humidified environment at 5% CO2 in air. Cells migrated under the agarose in a circle were fixed and stained. Diameters of the migration areas were measured with ocular micrometer.

Results

MIF reaction was maximum (44.2%) in the group IVEgESAg5 and minimum (10.8%) in the group IVASoAg1. The maximum MIF reaction was shown by eggs ES antigen and least by adult worm somatic antigen. The interesting observation was that migration inhibition increases as dose increased or we could say the reaction was dose dependent.

Conclusion

Increased value of MIF response in vaccinated mice suggested the involvement of lymphocytes in cell-mediated immunity. This study also proves that excretory-secretory (ES) antigen of eggs from Trichuris muris was more effective in imparting immunity in mice.     

Hair Contamination of Sheepdog and Pet Dogs with Toxocara canis Eggs

Background

We tried to investigate the hair contamination of pet dogs and farm sheepdog with Toxocara eggs in terms of the different sex and age groups in north-west of Iran (Urmia and its suburbs).

Methods

Hair samples were collected from a total of 138 pet and farm sheepdogs from November 2008 to June 2009 in Urmia City and the suburb (West Azerbaijan-Iran) and examined for the presence of T. canis eggs.

Results

T. canis eggs found in 60 samples altogether (pet and shepherd dogs) showed a contamination rate of 36.2%. The number of observed T. canis eggs in each microscope field was varied from 1 to > 400. The age of the dog was found a significant factor to influence the prevalence and intensity of contamination, with 82% of all the eggs recovered from puppies (six months and younger). Additionally, the numbers of eggs in farm sheepdogs were significantly higher than pet dogs (P<0.05).

Conclusion

This report shows that direct contact with T. canis infected dogs, particularly puppies from shepherd dogs, may pose a serious hazard to human. Besides, as they may harbor a considerable number of eggs on their hair, they can contaminate the soil and the environment.     

Toxocara Nematodes in Stray Cats from Shiraz,Southern Iran: Intensity of Infection and Molecular Identification of the Isolates

Background

Toxocara is a common nematode of cats in different parts of Iran. Despite the close association of cats with human, no attempt has been done so far for molecular identification of this nematode in the country. Therefore, current study was performed on identification of some isolates of Toxocara from stray cats in Shiraz, Fars Province, Southern Iran, based on morphological and molecular approaches, and also determination of intensity of infection.

Methods

This cross-sectional study was carried out on 30 stray cats trapped from different geographical areas of Shiraz in 2011. Adult male and female worms were recovered from digestive tract after dissection of cats. Morphological features using existing keys and PCR-sequencing of ITS-rDNA region and pcox1 mitochondrial l gene were applied for the delineating the species of the parasites.

Results

Eight out of 30 cats (26.7%) were found infected with Toxocara nematodes. All the isolates were confirmed as Toxocara cati based on morphological features and the sequence of ribosomal and mitochondrial targets. Intensity of infection ranged from one to a maximum of 39 worms per cat, with a mean of 10.25±12.36, and higher abundance of female nematodes.

Conclusion

The most prevalent ascaridoid nematode of stray cats in the study area was T. cati and female nematodes were more abundant than that of males. This issue has important role in spreading of eggs in the environment and impact on human toxocariasis.     

The Prevalence of Cryptosporidium spp. in Children,Taiz District,Yemen

Background

This is the first work done on cryptosporidiosis among the children in Taiz, Yemen.

Methods

A number of 712 samples were collected from children of different ages (ranging from 1 month to 12 years) from Dec 2006 to Aug 2007. The collected samples were examined by Sheather''s sugar floatation and Modified Ziehl- Neelsen stain as well as ELISA methods. The test results were statistically analyzed by SPSS software.

Results

The overall positive percentage was 43.7%. The higher incidence (36.2%) was occurred in males while the lowest incidence (32.7%) was observed in females (r=0.876; P=0.001). The correlation between infected cases and the type of drinking water was r =0.121. Among the cases examined by ELISA (92 cases), 26.1% were infected. The correlation between seropositivity and gender was r=0.652 (P=0.031).

Conclusion

Cryptosporidium spp. is a significant pathogen among children at Taiz. Fresh water supplies, education, eating habits and domestic animals are considered the main sources for transmission of cryptosporidiosis.     

Prevalence and Molecular Identification of Cryptosporidium Spp. in Pre-Weaned Dairy Calves in Mashhad Area,Khorasan Razavi Province,Iran

Background

Cryptosporidium parvum is a zoonotic pathogen transmissible from a variety of animals to humans and is a considerable public health concern. Dairy cattle have been identified in numerous reports as a major source of environmental contamination with this pathogen. The aim of study was to detect and isolate the Cryptosporidium spp. from fecal samples of naturally infected pre-wean calves in the Mashhad area

Methods

Overall, 300 fecal specimens from 1 to 30 days pre-weaned calves were collected from 10 farms in the Mashhad area the capital center of the Khorasan Razavi Province, Iran and microscopically examined for Cryptosporidium spp. All infected samples were also analyzed using nested –PCR. A polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis of the small-subunit (SSU) rRNA gene was also used to detect and identify Cryptosporidium spp. in PCR- positive samples.

Results

Eighty five (28.3%) of the specimens were positive for Cryptosporidium spp. The prevalence of Cryptosporidium spp. in 8-14 days old and diarrheic calves were significantly higher than other groups. Restriction digestion of the PCR products by SspI, VspI restriction enzymes and sequence analysis revealed the presence of C. parvum bovine genotype in all isolates.

Conclusions

Our results suggest that pre-weaned calves are likely to be an important reservoir of zoonotic C. parvum.     

Genotyping of Toxoplasma gondii Isolates from Soil Samples in Tehran,Iran

Background

The protozoan parasite Toxoplasma gondii can infect any warm blooded nucleated cells. One of the ways for human infection is ingestion of oocysts directly from soil or via infected fruits or vegetables. To survey the potential role of T. gondii oocyst in soil samples, the present study was conducted in Tehran City, Iran.

Methods

A total of 150 soil samples were collected around rubbish dumps, children''s play ground, parks and public places. Oocysts recovery was performed by sodium nitrate flotation method on soil samples. For molecular detection, PCR reaction targeting B1 gene was performed and then, the positive results were confirmed using repetitive 529 bp DNA fragment in other PCR reaction. Finally, the positive samples were genotyped at the SAG2 locus.

Results

Toxoplasma DNA was found in 13 soil samples. After genotyping and RFLP analysis in SAG2 locus, nine positive samples were revealed type III, one positive sample was type I whereas three samples revealed mixed infection (type, I & III).

Conclusion

The predominant genotype in Tehran soil samples is type III.     

Detection and Molecular Characterization of Cryptosporidium species in Recreational Waters of Chaharmahal va Bakhtiyari Province of Iran using nested-PCR-RFLP

Background

The aim of this study was to detect and characterize Cryptosporidium spp. in water samples collected from recreational ponds of Chaharmahal va Bakhtiyari Province of Iran.

Methods

Thirty water samples were collected from November 2009 to May 2010. Each sample contained 10 liters of water. We used the SSU rRNA-based PCR-RFLP technique.

Results

Out of thirty samples examined, 6 (20%) were positive for different Cryptosporidium spp. Restriction pattern analysis showed that C. parvum has been the most prevalent genotype, followed by C. hominis and C. canis, respectively. In this area, the higher prevalence of C. parvum compared with other genotypes is consistent with the distribution of cattle.

Conclusion

Farm animals, particularly cattle are the main source of cryptosporidial contamination for recreational waters in this area.     

Cryptosporidium Spp. Infection in Human and Domestic Animals

Background

Cryptosporidium spp. is a coccidian parasite infected humans and animals. Prevalence rate of Cryptosporidium spp. infection associated with is some parameters such as sampling, age, season, country and contact to domestic animals. This study aimed to determine Cryptosporidium spp. Infection in humans and some animals in rural areas of Shushtar district from Khuzestan Province, south- west of Iran.

Methods

In this study, Stool specimens were randomly collected from 45 cattle, 8 buffalos, 35 calves, 22 turkeys, 3 sheep, 2 geese as well as 62 humans in different seasons selected from rural areas of Shushtar district located in Khuzestan in the south- west of Iran from August 2009 to April 2011. The collected stool samples were examined by modified Ziehl-Neelsen staining method.

Results

Altogether, 68/115 (59.1%) domestic animals and 9/62 (14.5%) of humans were showed Cryptosporidium spp. infection in the study areas.

Conclusion

In this study we found the high frequency of Cryptosporidium spp. infection in the studied areas.     

A Preliminary Parasitological Survey of Hepatozoon Spp. Infection in Dogs in Mashhad,Iran

Background

We attempted to determine the prevalence of Hepatozoon spp. infection in Mashhad, northeast of Iran, via blood smear parasitology.

Methods

The prevalence was investigated by examination of blood smear parasitology, using blood samples collected from 254 dogs (51 strays and 203 privately owned-dogs).

Results

Two stray dogs (2/51; 3.92%) and two privately-owned dogs (2/203; 0.98%) were infected with Hepatozoon spp. Therefore, as per blood smear parasitology, the prevalence of Hepatozoon spp. infection was 1.57% (4/254). Sixteen out of 254 dogs (6.29%) were infested with ticks; all of which were Rhipicephalus sanguineus. One of the dogs infected with Hepatozoon spp. exhibited ticks at the time of examination. Concurrent infection with Ehrlichia canis and Leishmania infantum was not detected in the four Hepatozoon spp. infected dogs.

Conclusion

This is the first epidemiological study on the prevalence of Hepatozoon spp. infection in dogs in Iran.     

Prevalence of Cryptosporidium spp. in Camels and Involved People in Yazd Province,Iran

Background

Although infection of dromedary camels with Cryptosporidium spp. is rare in Iran, it is considered a zoonotic threat to the keepers and herders of camels. Thus we investigated the prevalence of Cryptosporidium in these two hosts in Yazd Province, a semi-arid region in center of Iran.

Methods

This study was conducted during 4 seasons (winter 2008, summer 2009, winter 2009 and summer 2010). Fecal samples (n=200) were collected from live camels. Also, 100 abomasal mucosa and related fecal samples of the slaughtered camels were investigated. Stool samples from 100 individuals who were in persistent contact with camels were also obtained. After staining by modified Ziehl-Neelsen method, the prepared specimens were studied microscopically. Results were analyzed using SPSS 16.

Results

The rate of infection in feces and abomasal mucosa of camels were 20.33% and 12%, respectively. In addition, simultaneous fecal and mucosal infection was detected in 3 cases in winter. Statistical analyses showed no significant relation between infection and age of camels, as well as their sex and the season. Cryptosporidiosis in people who were in long-term contact with camels was also investigated microscopically by obtaining stool samples of 100 individuals (50 in summers, 50 in winters), 24 of them being infected with Cryptosporidium spp. The rate of infection was higher in winter than summer (16/50 compared with 8/50).

Conclusion

The prevalence of Cryptosporidium spp. in camels and involved humans in Yazd Province is relatively considerable and of public health importance.     

Isolation and Genotyping of Acanthamoeba Strains from Environmental Sources in Ahvaz City,Khuzestan Province,Southern Iran

Background

Acanthamoeba spp. are free-living amoebae commonly found in the environmental sources such as water, soil, and air. This ubiquitous amoeba is the causative agent of amoebic keratitis (AK). The objective of the present study was to investigate the presence of Acanthamoeba spp. in water and soil sources in Ahvaz City, Khuzestan Province, southern Iran.

Methods

In general, 110 samples of water and soil were taken from different localities of Ahvaz including agricultural canals, rivers, and swimming pools. Filtration and cultivation were carried out on non-nutrient agar medium (NNA). Axenic cultivation was performed for all of positive isolates. PCR analysis was conducted on positive samples. Sequencing was done for 15 PCR products. Genotypes were identified by Blast search and homology analysis.

Results

Acanthamoeba spp. was found in 43 (71.6%) of samples of water and 13 (26%) soil samples. Genotyping of 15 samples proved that Acanthamoeba belonged to T4 (86.6%), T2 (6.6%), and T5 (6.6%) genotypes.

Conclusion

TYI-S-33 medium could be better than PYG medium for Acanthamoeba axenic culture.     

Malaria Epidemiology in Mersin Province,Turkey from 2002 to 2011

Background

Malaria is an infectious disease caused by Plasmodium spp. with high morbidity and mortality in human in tropical and subtropical regions. In recent years, number of malaria cases has been significantly reduced because of fight with the disease in Turkey. This study intended to investigate the malaria epidemiology in Mersin Province from 2002 to 2011 using data from the provincial Public Health Directorate.

Methods

Over ten years, 303573 blood samples were taken from the people by active and passive surveillance methods and blood smears were prepared. Smears were stained with Giemsa and examined under the microscope.

Results

Totally, 73 people including 44 male and 29 female were positive in terms of Plasmodium spp. It was determined that P. vivax observed in 67 cases while P. falciparum in 6 cases. Cases were mainly observed in 15 to 44 years old range, showed an increase between June-September periods and a significant decrease after 2006. Out of the 73 malaria cases, 54 cases were from Mersin Province and 13 cases were imported from another province of Turkey. Six cases were transmitted from abroad.

Conclusion

These results provide information about malaria epidemiology in an endemic area in Turkey and contribute its prevention in Mersin Province.     

Prevalence of Zoonotic Intestinal Helminths of Canids in Moghan Plain,Northwestern Iran

Background

The present study was aimed to elucidate the status of intestinal helminth infections in canids of Moghan Plain, northwestern Iran.

Methods

Eighty-five intestine samples from dead or shot wild canids, 59 fecal samples from sheepdogs and 5 from red foxes were collected from 2006 to 2008 and examined in Parasitology department of Pasteur Institute of Iran.

Results

Generally, adult worms, larvae, and eggs of 13 species of various parasitic helminths were recovered. Necropsy examinations showed that 96.47% animals harbored at least one helminth species. The prevalence of different species in necropsy were Mesocestoides sp. 84.7%, Rictolaria spp. 55.3%, Macranthorhynchus hirudinaceus 45.9%, Toxocara canis 43.5%, Toxascaris spp. 35.3%, Joyeuxiella sp. 34.1%; hookworms; 22.4%, Taenia spp. 11.8%, Alaria spp. 2.4% and Dipylidium caninum 1.2%. Besides, eggs belonging to 10 species of parasitic helminths were identified in 46 fecal samples and generally, 30.9% of samples harbored eggs of at least one helminth species.

Conclusion

The high prevalence of various helminth infections among canids in Moghan plain and contamination of environment by helminths eggs may increase the risk of infection for native people.     

First Report of Vannellidae Amoebae (Vannella Spp.) Isolated From Biofilm Source

Background

Members of the Vannellidae family are free-living amoebae (FLA) distributed mainly in water and soil sources. The present study reports the first isolation of this genus in the biofilm source from hospital environment in Tehran, Iran.

Methods

Biofilm samples were collected from hospital environment. Cultivation was performed in non-nutrient agar covered with a heat-killed Escherichia coli. Cloning of the suspected amoebae was done. PCR amplification and Homology analysis using the Basic Local Alignment Search Tool (BLASTn) was performed to search for the most similar reference sequences.

Results

Microscopic examination showed numerous fan-shaped amoebae and peculiar cysts different to the usual shape of typical FLA. Sequence analysis of the PCR- product revealed that the suspected amoebae are highly homologous with Vannella spp. gene (99% identity and 100% query coverage) available in the gene bank database.

Conclusion

Although Vannella spp. is not proved to be pathogenic itself, but they are capable of harboring pathogenic intracellular organisms such as Microsporidian parasites. Thus, identification of such amoebae can be of clinical importance, as they could lead to transmission of other pathogens to human.     

Development of Sensitive Detection of Cryptosporidium and Giardia from Surface Water in Iran

Background

The protozoan parasites Cryptosporidium spp. and Giardia are known to occur widely in both raw and drinking waters. They are two of the causative agents of waterborne outbreaks of gastroenteritis throughout the world. In the present study, a PCR assay and FA were developed for detection of Cryptosporidium oocysts and Giardia cyst in environmental samples.

Methods

We have detected Cryptosporidium spp. oocysts and Giardia cysts in seeded and unseeded environmental water samples by PCR method. Water samples were spiked with oocysts (50, 100,300,500) and filtrated with a 1.2-µm pore size cellulose nitrate and follow by DNA extraction and purification by QIAamp DNA mini kit. Nested-PCR assay amplified an 850 bp fragment of 18s rRNA gene specific for Cryptosporidium and 435 bp fragment of glutamate dehydrogenase (GDH) target gene for Giardia. Also many river water from north of Iran, be checked by these methods.

Results

Cryptosporidium and Giardia DNAs were detected in seeded water sample and Giardia was detected in all 5 water samples from river in north of Iran by nested- PCR and FA. Also in one river water sample, Cryptosporidium was detected.

Conclusion

This protocol is effective for detection of these waterborne parasites in treated and untreated water samples. This study can also serve as a platform for further investigations and research water source in Iran.     

Application of the Reverse Line Blot Assay for the Molecular Detection of Theileria and Babesia sp. in Sheep and Goat Blood Samples from Pakistan

Background

The present study was designed to detect the presence of tick-borne parasites (Theileria and Babesia spp.) in 196 blood samples collected from apparently healthy sheep and goats from two provinces, Punjab and Khyber Pukhtoon Khwa, in Pakistan.

Methods

Reverse line blot (RLB) assay was applied for the parasitic detection by the amplification of hypervariable V4 region of the 18S ribosomal RNA (rRNA) gene. A membrane with covalently linked generic and species specific oligonucleotide probes was used for the hybridization of amplified PCR products.

Results

Parasites were detected in 16% of the ruminant blood samples under study. Two Theileria species, T. lestoquardi and T. ovis, were identified in samples. 25, of the total 32, infected animals were from Khyber Pukhtoon Khwa.

Conclusion

Sheep were more prone to tick borne haemoprotozans as 81% infected samples were sheep as compared to 19% goats (P > 0.001). Risk factor analysis revealed that male (P = 0.03), animals infested by ticks (P = 0.03) and herd composed of sheep only (P = 0.001) were more infected by blood parasites.     

Urinary Schistosomiasis in Urban and Semi-Urban Communities in South-Eastern Nigeria

Background

In view of the massive rural-to-urban migration in Nigeria, investigations on transmission of urinary schistosomiasis were carried out in urban and semi-urban communities in Nike Lake area of Enugu State, Nigeria.

Methods

Urine samples of school children were tested for micro-haematuria using reagent strips followed by microscopic examination for Schistosoma haematobium eggs. Water contact sites were also identified and sampled for snails.

Results

The overall prevalence of S. haematobium eggs in school children was 4.64%. The mean intensity of infection was 1.14 ± 0.41 eggs/10ml urine. Males had insignificantly higher prevalence and intensity of S. haematobium infection than females. The youngest age group (4-7 years) had no infection. The prevalence of micro-haematuria (6.2%) was higher than that of microscopy, and this correlated positively with prevalence (r = 0.65, P < 0.01) and intensity (r = 0.50, P < 0.01) of the infection. Potential intermediate host of human shistosome collected were: Bulinus globosus, B. senegalensis and Biomphalaria pfeifferi. However, only B. globosus shed cercariae of S. haematobium, with a snail infection rate of 0.73%. Transmission was in the dry season coinciding with the drying of wells.

Conclusion

The results revealed that urinary schistosomiasis is prevalent, and that B. globosus and not B. truncatus as previously reported is the main intermediate host of urinary schistosomiasis in this part of Enugu State.     

Superoxide Dismutase (SOD) Enzyme Activity Assay in Fasciola spp. Parasites and Liver Tissue Extract

Background

The purpose of this comparative study was to detect superoxide dismutase (SOD) activities in Fasciola hepatica, F. gigantica parasites, infected and healthy liver tissues in order to determine of species effects and liver infection on SODs activity level.

Methods

Fasciola spp. parasites and sheep liver tissues (healthy and infected liver tissues), 10 samples for each, were collected, homogenized and investigated for protein measurement, protein detection and SOD enzyme activity assay. Protein concentration was measured by Bradford method and SODs band protein was detected on SDS-PAGE. SODs activity was determined by iodonitrotetrazolium chloride, INT, and xanthine substrates. Independent samples t-test was conducted for analysis of SODs activities difference.

Results

Protein concentration means were detected for F. hepatica 1.3 mg/ ml, F. gigantica 2.9 mg/ml, healthy liver tissue 5.5 mg/ml and infected liver tissue 1.6 mg/ml (with similar weight sample mass). Specific enzyme activities in the samples were obtained 0.58, 0.57, 0.51, 1.43 U/mg for F. hepatica, F. gigantica, healthy liver and infected liver respectively. Gel electrophoresis of Fasciola spp. and sheep liver tissue extracts revealed a band protein with MW of 60 kDa. The statistical analysis revealed significant difference between SOD activities of Fasciola species and also between SOD activity of liver tissues (P<.05).

Conclusion

Fasciola species and liver infection are effective causes on SOD enzyme activity level.     

Detection and Identification of Toxoplasma gondii Type One Infection in Sheep Aborted Fetuses in Qazvin Province of Iran

Background

The aim of this study was to apply the nested-PCR and bioassay methods in detection and genotyping of Toxoplasma gondii infection in provided sheep aborted fetus samples from Qazvin Province of Iran.

Methods

Eighteen sheep aborted fetal samples were studied by nested-PCR-RFLP, histopathological observation and microbiological assay. Bioassay in mice was carried out by inoculating the brain samples intraperitoneally.

Results

The results demonstrated the frequency of 66% infected sheep aborted fetal samples with T. gondii type one. Although we could not isolate any parasite from inoculated mice even after three passages, but it was confirmed histopathologically formation of cyst like bodies in prepared mice brain sections.

Conclusion

The results of the performed nested-PCR and formation of brain cyst in inoculated mice exhibited that T. gondii type one infection might be considered as one of the major causative agents for abortion in ewes.