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.     

Hematologic and Clinical Aspects of Experimental Ovine Anaplasmosis Caused by Anaplasma ovis in Iran

Background

Anaplasma ovis infections can cause clinical symptoms in acute phase and lead to huge economic losses in flocks. The aim of the present study was to investigate the hematological and parasitological changes in experimental anaplasmosis in sheep with Iranian strain of A. ovis.

Method

Five male sheep without any blood parasite infection were selected. One hundred ml heparinized blood was collected from splenectomised sheep that showed 6% A. ovis parasitemia. Inoculums of 20 ml blood were administered intravenously to each test animal. Hematological, parasitological and clinical changes of experimental anaplasmosis were studied in 0-38 days post infection.

Result

Parasitemia was detected 3 days post infection and reached its maximum level on the day 12 of experiment in test animals. Then the parasitemia was declined, but the organism could be found persistently until the last day of study. The red cell counts, packed cell volume and hemoglobin concentration were decreased and mean corpuscular volume was increased significantly during the infection period. Reticulocytosis and basophilic stippling were also detected. No significant changes were observed in total and differential leukocyte count and animal body temperature.

Conclusion

Experimental A. ovis infection in sheep resulted in marked normocytic normochromic anemia at the beginning of the infection which became macrocytic normochromic by the development of the disease. There were negative correlations between parasitemia and RBC, PCV and Hb values, therefore hematological assessment can be considered as a practical diagnostic tool in ovine anaplasmosis.     

Species Identification of Birds Nasal Trichobilharzia in Sari,North of Iran

Background

Cercarial dermatitis is known as an endemic parasitic disease in North of Iran, a hypersensitive skin reaction to the penetration of nonhuman schistosome larvae into human skin. In recent studies in this region, final and intermediate hosts were recognized and Trichobilharzia was identified as the main causative agent of cercarial dermatitis in this region, but to date the parasite species haven''t been identified. Therefore this study was performed to species identification of nasal Trichobilharzia in infected birds for the first time.

Methods

A total of 45 Anas clypeata birds identified as final host, were collected from Sari in North of Iran and infected nasal tissues analyzed using molecular techniques. Genomic DNA was isolated by phenol/chloroform extraction method and ITS region of rDNA were amplified with specific primers its5Trem and its4Trem, then sequenced area were compared with existing records in GenBank.

Results

Twelve samples were infected with Trichobilharzia and results of PCR reaction indicated that all of them belonged to T. regenti. The sequence alignment of present work isolates and those deposited in GenBank showed differences in nucleotide sequences of repeat region in ITS1.

Conclusion

Trichobilharzia regenti is the most frequent parasite of Anatid birds in North of Iran. This corresponds to the distribution of this parasite along the flyway of migratory birds, which annually migrate from Siberia and northern countries of Caspian Sea to wintering areas in southern regions of it.     

Detection of Acanthamoeba and Toxoplasma in River Water Samples by Molecular Methods in Iran

Background:

Free-living amoebae such as Acanthamoeba species may act as carriers of Cryptosporidium and Toxoplasma oocysts, thus, may play an important role in the water-borne transmission of these parasites. In the present study, a loop mediated isothermal amplification (LAMP) method for detection of Toxoplasma and a PCR assay were developed for investigation of Acanthamoeba in environmental water samples.

Methods:

A total of 34 samples were collected from the surface water in Guilan Province. Water samples were filtrated with membrane filters and followed by DNA extraction. PCR and LAMP methods used for detection of the protozoan parasites Acanthamoeba and Toxoplasma respectively.

Results:

Totally 30 and 2 of 34 samples were positive for Acanthamoeba and Toxoplasma oocysts respectively. Two samples were positive for both investigated parasites.

Conclusion:

The investigated water supplies, are contaminated by Toxoplasma and Acanthamoeba (oo)cystes. Acanthamoeba may play an important role in water-borne transmission of Toxoplasma in the study area. For the first time in Iran, protocol of LAMP method was used effectively for the detection of Toxoplasma in surface water samples in Iran.     

Seroprevalence of Neospora spp. in Horses in North East of Iran

Background

Neospora caninum, an obligate intracellular protozoan parasite, is recognized as a major cause of abortion in cattle, while limited information is presently available on the seroprevalence of Neospora antibodies in horses’ worldwide. The aim of the present study was to determine serologic prevalence of Neospora infection in horses in Iran.

Methods

Sera from 150 horses from Mashhad suburb in Razavi Khorasan Province, northeast Iran were examined for antibodies to Neospora spp. using Neospora modified direct agglutination test (N-MAT).

Results

Antibodies to this parasite were detected in 45 (30%) of the examined serum samples. Thirty four percent of the samples had titer of 1:40 while then reduced to 30% when 1:80 serum dilution was applied as significant cut off titer.

Conclusion

This study is the first investigation carried out on the Neospora in horses in Iran and indicates that horses in Iran are exposed to this parasite.     

Echinococcosis/Hydatidosis in Ilam Province,Western Iran

Background

Hydatidosis is a zoonotic disease of global prevalence. It causes considerable health problems and economic losses throughout the world, including Iran. The objective of this study was to assess the current status of echinococcosis/hydatidosis in the province of Ilam (western Iran).

Methods

From April to September 2011, 65 stray dogs were collected from urban and rural areas of Ilam City. Parasites were isolated from the dogs and stained with carmine. A taxonomic study was carried out by measuring different parts of helminths. Meat inspection documents from slaughterhouses in Ilam were used to assess the prevalence of hydatidosis during a 3-year period in sheep, cattle, and goats. ELISA test was used to detect the presence of antibodies to hydatidosis in human sera. Clinical records from 2000 to 2010 of either treated or diagnosed patients from public hospitals of this province were reviewed.

Results

The prevalence of Echinococcus granulosus infection in stray dogs was 9%. A total of 81,726 animals were assessed for hydatidosis; 2.94% (2403 cases) had liver hydatidosis and 2.34% (1918 cases) had lung hydatidosis. Within a 10-year period, 140 patients (91 females and 49 males) were treated for hydatidosis. Of 1200 human sera, 2.25% (27 patients) were seropositive for hydatidosis.

Conclusion

Hydatidosis is endemic in Ilam Province especially in rural area. The health and economic losses caused by the disease are significant; thus, our efforts need to be focused on the control of this disease.     

Seasonal and Geographic Distribution of Cercarial Infection in Lymnaea gedrosiana (Pulmunata: Lymnaeidae) In North West Iran

Background

Trematodes are a diverse group of endoparasites which require molluscan and vertebrate animals as intermediate and definitive hosts in their life cycle. The present study was carried out to determine the diversity and geographic distribution of infection with trematodes''cercariae in the snail Lymnaea gedrosiana from north-west Iran.

Methods

A total number of 6759 Lymnaeidae snails were collected from 28 snail habitats; of these L. gedrosiana was the prevalent snail (74.37%) which examined for cercarial infection by shedding method.

Results

The overall infection rate was 8.03%. The most frequent trematodes cercariae in the snail were xiphidiocercariae (81.98%), furcocercariae (32.26%), echinostome cercariae (5.19%), and monostome cercariae (1.24%). The highest infection rate in L. gedrosiana (100%) was with echinostome cercariae from Golestaneh in autumn.

Conclusion

Due to the important role of pond snails in transmission of cercariae to fish as a source of zoonotic diseases, it is essential to estimate the distribution and abundance of the snails and the rate of their infection with different trematodes’ cercariae, and establish control programs in each region.     

High Seroprevalence of Toxoplasma gondii Antibody in HIV/AIDS Individuals from North of Iran

Background:

Toxoplasmosis in immunocompetent people is generally asymptomatic but in immunocompromised patients including HIV/AIDS, cancer patients, and organ transplant recipients, etc. it can lead to serious pathological problems. The objective of current study was to determine the seroprevalence of T. gondii IgG and IgM antibodies in HIV/AIDS patients using ELISA technique in Mazandaran Province, northern Iran.

Methods:

Overall, 82 serum samples (61 males and 21 females) were collected from HIV/AIDS patients in Mazandaran Provinces, in 2013. Sera were surveyed employing ELISA assay. Data were analyzed using Chi-Square or Fisher exact test. In addition, before sampling a questionnaire was filled out for each subject.

Results:

Overall seroprevalence of examined sera was 96.3% for IgG antibody but none of the sera shown IgM antibody against T. gondii. The seroprevalence of toxoplasmosis in males and females was 96.7% and 95.2%, respectively. An antibody titer of >1 IU/ml was considered as positive. Furthermore, none of the included variables statistically was significant.

Conclusions:

Seroprevalence of chronic (latent) toxoplasmosis in HIV/AIDS patients in Mazandaran Province is high compared to toxoplasmosis in general population. Consequently, the risk of acquiring Toxoplasma encephalitis in examined seropositive HIV/AIDS patients of Toxoplasma is high.     

Helminth Infections in Rattus ratus and Rattus norvigicus in Tehran,Iran

Background

The aim of this study was to determine intestinal and liver helminth infections in Rattus rodents in Tehran Iran.

Methods

Overall, 306 traps were put in 39 different regions in Tehran from 2009 to 2010. Rodents, including R. rattus and R. norvegicus were caught by live-traps. They become unconscious and the spinal cords were cut, afterwards the body was dissected and the stomach, small intestine, large intestine, liver, and cecum were studied separately. The dominant type and the prevalence rate of parasites in the rodents were determined based on the infected parts of their body.

Results

After recognition of the helminthes’ types, among the 120 total number of rodents, 39 belonged to males, while among the infected rodents, 57(47.5%) were female and 18(15%) were male. The prevalence of infection in Tehran was 62.5%. Seventy cases (58.33%) of helminth infections were observed in R. rattus and 5 cases (4.16%) were observed in R. norvegicus. The maximum prevalence (15.5%) was seen in the center and east part of Tehran, while the minimum (9.16%) was in the north part of the city. The helminthes types and the corresponding percentages were Hymenolepis nana fraterna (35.8%), Heterakis spumosa (17.5%), Hymenolepis diminuta (7.5%) and Capillaria annulosa (1.6%). The dominant rodent was Rattus rattus and among the identified helminthes, Hymenolepis diminuta and Hymenolepis nana fraterna are zoonotic ones.

Conclusion

The information presented here improves our understanding of the major parasitic infections that rodents harbor and can transmit to human and animal populations in Iran. To prevent infectivity of human, the hazard of the identified zoonotic species needs to be contemplated.     

An Epidemiological Survey of Setaria in the Abdominal Cavities of Iranian Sistani and Brahman Cattle in the Southeastern of Iran

Background

In this experiment, abdominal cavity of 518 Iranian Sistani cattle and 498 Brahman cattle were inspected for the presence of Setaria spp. from April 2012 - May 2013.

Methods

The species were determined by microscopic examination of the morphological characteristics of the anterior and posterior parts of the parasites and authentic guidelines.

Results

The overall prevalence of Setaria spp. was 28.6% and 36.5%, respectively and this difference was significant (P<0.05). Out of 148 Sistani cattle which were infected with Setaria, 51(34.4%) were infected with S. digitata, 31 (20.9%) were infected with S. labiatopapillosa, 65 (43.9%) showed mixed infection of S. digitata and S. labiatopapillosa and one case (0.6%) was infected with mixed infection of S. labiatopapillosa, S. digitata and S. marshalli. These values were 87 (47.8%), 27 (14.8%), 67 (36.8%) and 1 (0.5%) for 182 infected Brahaman cows, respectively. The proportion of infected cattle in spring and summer was greater than cooler season (autumn and winter) significantly (P<0.001). The prevalence of infection with Setaria in 2-3 years old Sistatni cattle (42.2 %) was greater than other age categories (P<0.05). Furthermore, the infection rate between males (25.5%) and females (37.3%) Iranian Sistani cattle showed significant difference (P =0.009).

Conclusion

It is important to point out the presence of cerebrospinal setariosis, namely in sheep, goats and horses in the investigated area.     

Occurrence of Thermotolerant Hartmannella vermiformis and Naegleria Spp. in Hot Springs of Ardebil Province,Northwest Iran

Background

Geothermal waters could be suitable niches for thermophilic free living amoebae including Naegleria and Hartmannella. Ardebil Province, northwest Iran is popular for having many hot springs for recreational and health purposes activity. The present research is the first molecular based investigation regarding the presence of Naegleria and Hartmannella in the hot springs of Ardebil Province in Iran.

Methods

Overall, 30 water samples were taken from waters of thermal hot springs in Ardebil Province, Iran during 2010-2011. All collected samples were transferred to Dept. of Parasitology and Mycology, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Cultivation of concentrated water samples was performed using culture-enrichment method. Cloning of the target amoebae was obtained and morphological and molecular analysis was done using page key combined with two sets of primers, respectively. Sequence analysis and homology search was used for strains identification.

Results

Of 30 water samples, 8 (26.7%) were positive for thermotolerant Vahlkampfiids and Hartmannella based on morphological characteristics of vegetative form and double walled cysts. Cloning of the target amoebae were done successfully. Sequencing of the positive isolates revealed that the strains belonged to Naegleria (N. carteri and N. spp) and H. vermiformis.

Conclusion

The result highlights a need for improved filtration and disinfection and periodic monitoring of recreational thermal waters in order to prevent disease related to free- living amoebae. This is the first comprehensive molecular study of thermophilic Naegleria and Hartmannella in hot springs of Iran.     

Occurrence of Nasal Nosocomial Myiasis by Lucilia sericata (Diptera: Calliphoridae) In North of Iran

We report a case of human nasal nosocomial myiasis in a 69-year-old rural man with numerous live maggots in nostrils. The patient was admitted in Emergency Ward due to dyspnea. After 72 hours the companion of patient discovered larva. The presence of the third instar larva indicated that the infestation was not more than three days. The collected instars were cleared, fixed and morphological studies then were carried out precisely and the larva was identified as Lucilia sericata.     

Histopathologic Effects of Dirofilaria immitis Microfilaria on Internal Organs of Dog Confirming by PCR Technique

Background

The heartworm disease is an infectious disease of dogs with Dirofilaria immitis combined with cardiovascular and circulatory abnormalities. The heartworm disease can become a serious health risk when associated with a severe infection. In this study, a male, 8 year-old dog that died suddenly was necropsied and all tissues were examined grossly.

Methods

Major organs including heart, lungs, liver, spleen, kidneys, brain, eyes, and testis were fixed in 10% neutral formalin, embedded in paraffin, sectioned at 5-µm thickness, stained with hematoxylin and eosin, and examined with a light microscope. For each examined organ, paraffin-embedded tissues were cut and placed in eppendorf tubes for genomic DNA extraction. PCR was performed using two sets of primers for amplification of a 302 bp ITS-2 gene fragment and a 203 bp cytochrome oxidase subunit 1 (CO1) gene fragment of D. immitis.

Results

During the necropsy examination, 46 adult D. immitis were found in the portal vein, right ventricle, and atrium of the heart and pulmonary trunk. Microscopically, microfilarias were found throughout the vessels of different organs including lungs, kidneys, liver, heart, brain, and spleen. All tissues examined by PCR were positive for D. immitis ITS-2 and CO1.

Conclusion

PCR technique now represents an effective method for identification of D. immitis from formalin-fixed samples.     

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.     

Malaria in Mazandaran,Northern Iran: Passive Case Finding During 1997-2012

Background

Malaria is one of the most important parasitic diseases in tropical and temperate regions. The aim of this study was to determine the trend of malaria in Mazandaran Province, northern Iran during 1997-2012.

Methods

This retrospective study was conducted from 1997 to 2012. The population''s study was individuals who registered at health centers of Mazandaran Province. Peripheral blood smear were prepared for each case, stained with Giemsa and examined by light microscope. In addition to demographic data, other parameters including Slide Positive Rate (SPR), Annual Parasite Incidence (API) and Annual Blood Examination Rate (ABER) were analyzed.

Results

In total, 844 cases of malaria were reported. Plasmodium vivax was predominant species with 821 cases (97.4%). The number of malaria cases increased from 1997 to 2005 and then decreased to 3 cases in 2011. Some cities had not reported any cases during last three years. The highest infection rate, 163(20.07%), was seen in 2001-02. The SPR had the highest value (0.54%) in 2004-05. The maximum API and ABER were observed in 2001-02 and 1997-98. 641(75.9%) of cases were imported from hyperendemic areas such as Afghanistan and South-eastern Iran and 94 (11.1%) malaria patients were recorded as introduced cases. The highest infection rate of malaria (21.3%) was seen in Babolsar.

Conclusion

Extensive malaria control should be continued to Mazandaran to become malaria-free region and in prevention of re-introduction stage.     

Radular Teeth Morphology in Limax (Caspilimax) keyserlingi (Martens, 1880) and Parmacella ibera (Eichwald, 1841) from Northern Iran

Background

Slugs have been known worldwide as important pests of agricultural and horticultural production. They also play a role as intermediate or definitive hosts of helminths parasite. In this purpose, current study was carried out to examine slug radular teeth structure and slug infection with helminths larvae in north of Iran.

Methods

A total number of 114 slugs were collected from center and east parts of Mazandaran province from May 2011 to June 2012. The specimens were rinsed, measured, and identified. The radula of all collected slugs was extracted and stained by using Mallory II. For detecting the helminths parasite infection, foot- head and viscera of examined slugs were removed, minced, and digested with 4.5% acid pepsin.

Results

Two species of Limax (Caspilimax) keyserlingi (Martens 1880) (11.4%, 13/114) and Parmacella ibera (Eichwald 1841) (88.6%, 101/114) were prevalent in the region. There was significant difference between body length and shell size. P. ibera had the highest number of teeth rows (145±5). The radular teeth formula was approximately similar in both identified slugs. In P. ibera, there was no significant difference in the average length and width of radula. The radular teeth in L. keyserlingi were larger and thicker than P. ibera. In all examined slugs for helminths larvae infection, P. ibera (7.69%, 1/13) was infected with Strongyloid larvae from Fereidonkenar area.

Conclusion

Two prevalent species of slugs exist in the same region of which P. ibera has capability to play a role as intermediate host of nematode helminths. Radular morphology within the slug species may be also systemically informative.     

Molecular Characterization of Echinococcus granulosus from Hydatid Cysts Isolated from Human and Animals in Golestan Province,North of Iran

Background

The aim of the present study was to determine the molecular characteristics of Echinococcus granulosus from paraffin-embedded tissues of hydatid cysts isolated from human and protoscoleces of hydatid cysts from sheep, cattle and camel isolates using PCR- RFLP of ITS1- rDNA analysis in Golestan Province, northern Iran.

Methods

E. granulosus isolates from human patients infected with hydatid cyst and protoscoleces from hydatid cysts of sheep, cattle and camel isolates were collected from different hospitals and the abattoir throughout the Golestan Province. In all, 60 E. granulosus genomic DNA were extracted and examined by PCR - ITS1 of rDNA and amplified using BD1 / 4S and EGF1 / EGR2 primers, followed by RFLP using Alu1, Msp1 and TaqI restriction enzymes.

Results

The PCR-ITS1 products obtained from sheep, cattle and human isolates were similar to sheep strain (1000 bp and 391 bp). Majority of the camel samples yielded 295 bp DNA bands. RFLP -ITS1 of E. granulosus with Taq1 in human, sheep and cattle isolates showed similar patterns in the number and size of DNA. RFLP methods in camel isolates showed a different genotype, using Taq1, whereas no DNA bands were observed using Alu1 in camel and human isolates. Therefore, two clearly distinguishable banding patterns of E. granulosus were obtained with the three enzymes, which separating human, sheep and cattle isolates from the camel origin.

Conclusion

The results indicate the possible of transmission of the G1 and G6 genotypes of E. granulosus between livestock animals and human in Golestan Province.