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.     

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.     

Epidemiological Aspects of Canine Dirofilariasis in the North of Iran

Background

Dirofilaria immitis is an important parasite in dog and other carnivores. Our objective was study on incidence and periodicity of heartworm in north of Iran and using other methods for its diagnosis in addition to Parasitology exam.

Methods

This survey spanned two years, between 2006 and 2008. Blood samples were collected from 431 stray dogs distributed along north of Iran, the coastal areas of the Caspian Sea. The Knott''s modified test was used for diagnosis of D. immitis and other filariae. Meanwhile, the periodicity of microfilaria in peripheral blood circulation was calculated and the imaging diagnosis techniques of four dogs that had positive results were done.

Results

Diagnostic parasitology results indicated that 16.01% of stray dogs were microfilaremic. Two different microfilariae were diagnosed: D. immitis in 13.69%, Dipetalonema reconditum in 1.86% and in 0.46% both of them. There was no statistically significant between infection to fiariae with sex and age of dogs. Also study on the periodicity of the presence of microfilaria in peripheral circulation showed that the highest rate of those was at 1 am and the lowest rate at 12 pm. Radiographic study showed distinctive signs with varied degrees of severity included: Tortuous and enlargement of main and lobar pulmonary artery, pulmonary parenchymal lesions and Right side heart enlargement that confirmed in electrocardiography. Also in echocardiographic images observed short parallel-sided images with the appearance of equal signs that indicated the presence of the heartworm.

Conclusion

These results showed that to obtain a reliable diagnosis of heartworm infection, imaging tests could support parasitological exams.     

Transovarial Transmission of Babesia ovis by Rhipicephalus sanguineus and Hyalomma marginatum

Background

Rhipicephalus sanguineus and Hyalomma marginatum are the most common species in sheep herds in Northeast of Iran. There is preliminary evidence that these species may be the vectors of Babesia ovis in Iran. We carried out two experiments in Mashhad area, Khorasan Razavi Province to determine whether B. ovis could be transovarially transmitted by R. sanguineus and H. marginatum.

Methods

In experiment 1, adults of laboratory reared H. marginatum and R.sanguineus were infected with B. ovis isolated from naturally infected sheep in Mashhad area by feeding the ticks on the sheep inoculated intravenously by infected blood samples. The inoculated sheep showed clinical signs with parasitaemia while the adult ticks were engorging on them. The engorged females were collected and kept at 28°C and 85% relative humidity in incubator. Then, larval, nymphal and adult stages derived from engorged females were used to infest the clean sheep. In experiment 2, two splenectomized sheep were infested only with the same adult ticks of two species.

Results

Examination of smears and PCR of blood samples to detect of B. ovis in infested sheep in two experiments were negative.

Conclusion

It seems that R. sanguineus and H. marginatum can not transovarially transmit B. ovis in sheep.     

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.     

Prevalence and Rate of Parasitemia of Haemoproteus columbae in Columba Iiviadomesticain Southwest of Iran

Background

Parasites affect the health and productivity of birds. Haemoproteus columbae occurs in pigeons widely in tropical and subtropical regions. The present investigation was concentrated on the prevalence of H. columbae and rate of parasitemia in domestic pigeons in southwest of Iran.

Methods

Pigeons regimented in three groups, less than six months old, between six and twenty four months old and more than twenty four months old.Then stained blood smears were studied for presence of H. columbae and finally rate of parasitemia in every group calculated.

Results

Mature and immature stages of H. columbae gametocytes were found in 24% of blood smears prepared from 100 healthy domestic pigeons. Mean of parasitemia in infected pigeons was 9.58%. Mean size of macrogametocytes was 4μm×15μm and mean size of microgametocytes was 3μm×12μm. Mean of parasitemia in infected females was more than males and pipers. Mean of parasitemia in infected old pigeons (pigeons with more than twenty four months old) was more than pigeons with less than six months old and pigeons between six and twenty four months old.

Conclusion

This study show the prevalence and rate of parasitemia in domestic pigeons in southwest of Iran. We should be care about this parasite in pigeons by knowing the prevalence and high risk groups.     

Trichinella Infection in Wildlife of Northeast of Iran

Background

The objective of this investigation was to detect the presence of Trichinella in some carnivores of Mashhad in northeast of Iran and to identify Trichinella species circulating in this area.

Methods

The present study was carried out using muscle tissue collected from 120 stray dogs, 26 wild boars, 25 rodents, two foxes and two hyenas captured in Mashhad City, province of Khorasan Razavi, Iran.

Results

Trichinella larvae were detected in three stray dogs by artificial digestion and compression. All larvae were identified as T. britovi using multiplex PCR.

Conclusion

This is the first report of identification of T. britovi in stray dog in Iran.     

Canine Heartworm in Southeastern of Iran with Review of disease distribution

Background

Heartworm (Dirofilaria immitis) is mosquito-borne filarial nematode capable of causing serious cardiopulmonary disease in canines and felines, and pulmonary dirofilariasis in man. This research was conducted with the objectives of determining the incidence and assessing possible risk factors of canine heartworm in the southeast of Iran.

Methods

From October 2012 to September 2013, blood samples from 87 dogs from Zabol area in Sistan and Baluchestan and 33 dogs from Bam area in Kerman Province were examined for detection of Dirofilaria immitis using modified knott test and serology.

Results

Out of 120 dogs, 29 (24.2%; 95%CI: 16.6-31.8%) were positive, serologically. The overall seroprevalence of D. immitis in dog in Zabol and Bam was 27.5% (95% CI: 24.7-32.5%) and 15.15% (95% CI: 12.3-20.7%), respectively. 28.8% of stray dogs and 20.6% of housed dogs in the study areas were seropositive. Seroprevalence of D. immitis was not significantly different between stray and housed dogs (P = 0.295). Investigation of seasonal dynamic of infection with D. immitis in stray and housed dog showed that the proportion of infected dog in spring and summer was greater than colder season (autumn and winter) which was not significant. The prevalence of infection with D. immitis in >5 years old stray dogs (53.8%) was greater than other age categories while in housed dogs infection rate was greater in 3-5 years old (27.3%).

Conclusion

It is important to point out the increased incidence of canine heatrworm in Iran. In order to stop the spread of canine heartworm, preventive measures must be taken now.     

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.     

Dientamoeba fragilis in Gaza Strip: a Neglected Protozoan Parasite

Background

The aim of this study was to detect Dientamoeba fragilis by iron haematoxylin stain, as well as its prevalence, and association between D. fragilis infection and diarrhoea among patients attending Al-Nuseirate Refugee Camp Clinic, Gaza Strip.

Methods

A cross-sectional study was conducted among 319 children and adults with age ranges from (1 to 75) years old, attending Al-Nussirat Clinic, and who were complaining from clinical symptoms, like diarrhoea and abdominal pain.

Results

28 individuals were infected with D. fragilis with a prevalence of 8.8%. The detection of 28 cases infected with D. fragilis was proved using iron haematoxylin stain, but no case was detected by direct smear or formal-ether sedimentation technique. The most frequent symptoms were abdominal pain (96.4%) and diarrhoea (71.4%) in patients with diantamoebiasis and this was statistically significant (P= 0.03). Co-infection between D. fragilis and Entamoeba histolytica/dispar was 50% and between D. fragilis and Giardia lamblia was 7.1%.

Conclusion

D. fragilis was present in the patients stool samples and was detected and proved using iron haematoxylin stain.     

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.     

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.     

Molecular Study of Sheep Malignant Theileriosis at Barka Region in the Sultanate of Oman

Background

We used the PCR technique based on the abovementioned primer pair and sequencing to demonstrate the Theileria infection in the sheep samples collected from Sultanate of Oman.

Methods

According to the frame work of “integrated control of ticks and tick borne diseases in globalized world managed by EU-ICTTD-3 project, the samples from blood, liver, spleen, lymph node and lung were sent to the laboratory of Iranian Research Center for Ticks and Tick-borne Diseases (IRCTTD). Samples from blood smear and impression smears from liver, spleen, lymph node, and lung were analyzed by Geimsa staining. The DNA was extracted from the abovementioned samples and analyzed by PCR technique using specific primers derived from the nucleotide sequences of 18S rRNA gene of T. lestoquardi, which can amplify the common region in other Theileria and Babesia spp. Subsequently the amplified DNA was sequenced.

Results

The analysis of blood smears of the sheep was negative for piroplasmosis performed through the Giemsa staining. The impression smears prepared from liver, spleen, lymph node, and lung showed suspicious structures mimicking Theileria schizonts in some cells. The results showed an expected PCR product of 428 bp in length, which is specific for Theileria spp. The PCR products were subsequently sequenced. The corresponding nucleotide sequence is registered under accession number {"type":"entrez-nucleotide","attrs":{"text":"JF309152","term_id":"323695911","term_text":"JF309152"}}JF309152 in GenBank. The sequence alignment in GenBank showed that the PCR products had 99% homology to the known T. lestoquardi registered under accession number {"type":"entrez-nucleotide","attrs":{"text":"AF081135","term_id":"7576455","term_text":"AF081135"}}AF081135 in the GenBank.

Conclusion

Oman sheep are highly susceptible for Theileria infection and the infected sheep mostly die before the microschizonts or erythrocytic form of Theileria appears in the nucleated or erytrocytic cells respectively.     

Prevalence of Linguatula serrata Nymphs in Mesenteric Lymph Nodes of Cattle and Buffaloes Slaughtered in Ahvaz Abattoir,Iran

Background

Linguatula serrata, one of the parasitic zoonoses, inhabits the canine respiratory system (final hosts). The objective of this study was to determine the prevalence rate of L. serrata nymphs in mesenteric lymph nodes (MLNs) of cattle and buffaloes (intermediate hosts) that were processed in the Ahvaz, Iran abattoir.

Methods

During November 2010 to March 2011, 223 animals (119 cattle and 104 buffaloes), in different sex and three age groups (<2, 2–< 3 and 3-> 3 years old) were sampled randomly at Ahvaz abattoir. Up to 35 grams of their mesenteric lymph nodes were examined separately for nymphal stages of L. serrata by digesting the samples with acid- pepsin method, collected the nymphs and counted under stereomicroscope.

Results

Overall 37(16.6%) of 223 animals were infected with L. serrata nymphs in their mesenteric lymph nodes. Prevalence of the infection in cattle and buffaloes were 16.8% and 16.3% respectively. The number of collected nymphs of MLNs was ranged from 1 to 16. No significant differences were seen in the infection rates between males and females (sexes) and age groups in the cattle and buffaloes (P <0.05).

Conclusion

Linguatula serrata has an active life cycle in the studied area and a zoonotic potential for transmission between animal and human. Avoiding use of raw MLNs to dogs can help reduce the infection.     

Molecular Diagnosis of Trichomoniasis in Negative Samples Examined by Direct Smear and Culture

Background

Trichomoniasis is an extremely common sexually transmitted infection (STI) worldwide and is associated with important public health problems, including amplification of HIV transmission. This disease is in forms of symptomatic and asymptomatic in women and may depend on host as well as parasite variables. Most of the studies reported from females are based on examination of vaginal secretions and urine samples by direct smear and culture in modified Diamond''s media. The aim of this study was checking the samples, which were negative by direct smear and culture, with PCR technique.

Methods

The urine samples and vaginal discharge of patients attending Gynecology Clinics of Mazandaran Province, Iran with different symptoms rechecked for Trichomonas vaginalis by PCR technique using primers targeting a conserved region of the beta-tubulin genes of the parasite. Data were analyzed by Epi Info software program

Results

Out of 161 negative samples by direct smear and culture, seven samples (4.3%) were positive by PCR technique.

Conclusion

Diagnosis of trichomoniasis by PCR is a sensitive and specific method that could play important role to help the physicians for properly treatment and control of infection.     

Phylogenetic Analysis of Theileria annulata Infected Cell Line S15 Iran Vaccine Strain

Background

Bovine theileriosis results from infection with obligate intracellular protozoa of the genus Theileria. The phylogenetic relationships between two isolates of Theileria annulata, and 36 Theileria spp., as well as 6 outgroup including Babesia spp. and coccidian protozoa were analyzed using the 18S rRNA gene sequence.

Methods

The target DNA segment was amplified by PCR. The PCR product was used for direct sequencing. The length of the 18S rRNA gene of all Theileria spp. involved in this study was around 1,400 bp.

Results

A phylogenetic tree was inferred based on the 18S rRNA gene sequence of the Iran and Iraq isolates, and other species of Theileria available in GenBank. In the constructed tree, Theileria annulata (Iran vaccine strain) was closely related to other T. annulata from Europe, Asia, as well as T. lestoquardi, T. parva and T. taurotragi all in one clade.

Conclusion

Phylogenetic analyses based on small subunit ribosomal RNA gene suggested that the percent identity of the sequence of Iran vaccine strain was completely the same as Iraq sequence (100% identical), but the similarity of Iran vaccine strain with other T. annulata reported from China, Spain and Italy determined the 97.9 to 99.9% identity.     

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.     

Molecular and Seroepidemiological Survey of Visceral Leishmaniasis among Humans and Domestic Dogs in Mazandaran Province,North of Iran

Background

New cases of visceral leishmaniasis (VL) have been reported recently in some parts of Mazandaran Province, north of Iran where the first human case of VL was reported in 1949. This study aimed to determine the present status of Leishmania infantum infection among humans and domestic dogs using serological and molecular methods in central parts of Mazandaran Province.

Methods

In this cross-sectional study, blood samples were randomly collected from 402 humans and forty-nine domestic dogs throughout 2009 and 2010 in the central part of Mazandaran Province including Semeskadeh and Kiakola districts where recent cases of human visceral leishmaniasis had been reported there. All the collected samples were tested by direct agglutination test (DAT) for the detection of anti-Leishmania infantum antibodies as well as convenience PCR assay on whole blood samples for detection of leishmanial infection and identification of Leishmania species.

Results

None of 402 collected human (402) and dog (49) blood samples showed anti Leishmania infantum antibodies at titers 1:3200 and 1:320 as cut-off values of DAT, respectively but only 2 of domestic dogs (4.1%) were found PCR-positive corresponding to L.infantum.

Conclusion

This study confirms the circulation of L. infantum at least among domestic dogs and highlights the sporadic pattern of VL in the studied areas. Further investigations regarding to sand flies fauna and wild canines as reservoir hosts of the disease, are recommended.