Secretory Excretory and Somatic Immunogenic Antigens Profiles of Adult Fasciola spp.

Fascioliasis is a common human-animal disease that is reported in most parts of the world. Fascioliasis is also prevalent in different provinces of Iran. Since it has done no study on the excretory/secretory and somatic immunogenic antigens profiles of adult Fasciola in Iran, the present study was performed on the Fasciola spp. collected from Mazandaran province. For this purpose, the Fasciola worm was isolated from the liver of infected sheep, then its excretory/secretory and somatic antigens were prepared from adult worms. The protein of the samples was measured by the Lowry method. Then, somatic and secretory excretions were examined by SDS-PAGE and the protein profile of the two substances was determined. To evaluate the immunogenicity, the somatic and secretory excretions antigens of Fasciola spp. were injected into white rabbits and after boosting, the blood serum of the rabbits was collected and then Western blotting was performed on them and the results were evaluated. According to the results of Western blotting, 11 somatic antigen bands with a molecular weight of 149, 122, 99, 85, 75, 65, 50, 46, 40, 37, 30 kDa and 12 protein bands of excretory/secretory antigens with molecular weights of 100, 82, 75, 70, 58, 55, 47, 40, 38, 37, 30,25 kDa were observed in adult Fasciola spp. that immunogenic, which appear to have a protective effect or can be used to prepare a diagnostic kit.

Molecular Identification of Cutaneous Leishmaniasis Vectors in Alborz Province, North of Iran.

Cutaneous leishmaniasis (CL) is a vector-borne disease widely distributed in tropical and subtropical areas of North and South America, Europe, Asia, and Africa. Considering the increasing number of CL cases in recent years and the fact that no study has been conducted to identify CL fauna and vectors in Alborz province, this study was carried out to identify sand flies and CL vectors in this region. Sand flies were collected from August to October 2021 from plain and mountainous indoor and outdoor areas of the region using sticky paper traps and were detected morphologically. DNA was extracted from the midguts of female sand flies. In this study, 1157 sand flies were collected and identified. The number of sand flies caught from indoor and outdoor places was 367 (31.72%) and 790 (68.28%), respectively. Overall, six species of flies were of the genus Phlebotomus (Raynal, 1937), including Phlebotomus papatasi (P. papatasi, 695 [60.07%]; Scopoli, 1786), P. kandelakii (13 [1.12%]; Shchurenkova, 1926), P. sergenti (232 [20.05%]; Parrot, 1917), P. major (14 [1.21%]; Annandale, 1910), P. caucasicus (4 [0.35%]; Marzinowsky, 1917), P. alexandri (18 [1.56%]; Alexandri Sinton, 1920), and four were of the genus Sergentomyia (Artemiev, 1978), including Sergentomyia tiberiadis (109 [9.42%]; Adler, Theodor & Lourie, 1930), Sergentomyia baghdadis (53 [4.58%]), Sergentomyia sintoni (14 [1.21%]; Sintoni Pringle, 1933), Sergentomyia clydei (5 [0.43%]). P. papatasi spp. were dominant in indoor and outdoor places, with a prevalence of 695 (60.07%). The Leishmania major (L. major) gene was identified in five samples of P. papatasi spp. This suggests that P. papatasi is the potential vector spp. in the study area. Moreover, L. major was confirmed as the aetiological agent of CL cases in Alborz province. The identification of vectors and parasite spp. is very important for the treatment and operational planning of disease vectors.

Prevalence and Early Detection of Hypodermosis in Goats using a Competitive ELISA System in Lorestan, Iran.

This study aimed to determine the prevalence and early detection of hypodermosis in goats by the investigation of Przhevalskiana larvae and sera collected from the infested animals. This study was conducted in Lorestan province, located in the South-West of Iran, from April 2017 up to April 2018. A total of 3350 goats slaughtered in Lorestan abattoirs were investigated by clinical-parasitological examinations in different periods. The larvae were collected from the back and flank regions of the slaughtered goats. The number of infested animals, gender and age, number of maggots present on the body of each animal, location, and larval stage of warble flies were recorded in this study. To detect an infestation in the early period, a total of 150 blood samples were randomly collected from the field animals in Lorestan, Iran. The morphological findings showed that out of 3350 goats examined, 706 (21.07%) goats were infested. Furthermore, three species of Przhevalskiana, including P. Silenus (n=726, 50.07%), P.crossii (n=440, 30.43%), and P. aegagri (n=284, 19.59%) were recognized as the causative agents of goat hypodermosis in this province. No significant difference was observed between genders and/or among the age groups (P>0.05). The anti-Przhevalskiana antibodies in the serum samples were detected using ELISA from August up to mid-September (summer). Clinical diagnosis of infestation was usually performed from late October until mid-March (winter) by visual observations and direct palpation of warbles in the back and flank regions of the animals. It could be concluded that the use of ELISA can help to detect hypodermosis among goats in the early stages.

Development of Nano-ELISA Method for Serological Diagnosis of Toxoplasmosis in Mice.

Toxoplasmosis is a widespread parasitic disease caused by a protozoan parasite Toxoplasma gondii. Currently, nanotechnology has been used for the diagnosis of many infectious diseases. It could be due to the fact that nanoparticles play an important role in accurate and fast diagnosis. The purpose of this study was to design a Nano-enzyme linked immunosorbent assay (Nano-ELISA) kit using excreted/secreted (E/S) antigens to have higher sensitivity and specificity than those reported for the designed enzyme-linked immunosorbent assay (ELISA) kit for the diagnosis of Toxoplasmosis in mice. Firstly, the serum samples were collected from 15 infected mice with T. gondii and 15 healthy ones. Then, E/S antigens were separated from parasite tachyzoites and used for designing an ELISA kit. In addition, the mice sera were evaluated using the designed ELISA kit. Finally, the serum samples were assessed by Nano-ELISA kits designed with E/S antigen and conjugate of gold nanoparticles. The obtained results of the present study showed that the sensitivity and specificity of the designed ELISA kit were reported as 80% and 86.66%, respectively, that both improved to 93.33% in these sera with the designed Nano-ELISA kit. This finding revealed the significant improvement of sensitivity and specificity using gold nanoparticles in designing the ELISA kit. Furthermore, according to the literature, the use of E/S antigens in designing recognizable ELISA kits has been always highlighted considering the presence of numerous antigens in T. gondii. The results of this study revealed that the use of E/S antigens in the preparation of an ELISA kit was very effective. This is very important, especially in the lower titers of antibody requiring a more accurate diagnosis. On the other hand, the Nano-ELISA method designed with E/S antigens can be more sensitive and specific than ELISA for the diagnosis of Toxoplasmosis and can be the basis for further studies in this regard.

Serodiagnosis of Przhevalskiana spp. Infestation in Goats Using a Competitive ELISA.

Goat warble-fly infestation is one of the parasitic diseases caused by the larvae of przhevalskiana spp. It is known to be a major challenge for health and welfare in infested goats and causes high economic losses in livestock worldwide. The detection of goat grub was previously conducted by direct palpation of second and third stage larvae in the back and flank site of the live animals or visual evaluation of the carcasses in slaughterhouses. However, due to the small size of the first instar larvae of przhevalskiana (less than 1 mm during emerging from the egg), some of the infected cases were ignored and recorded as negative samples. Immunodiagnostic procedures as easy and cost-effective diagnostic methods provide early detection of myiasis in living animals (even when the larvae are still migrating or are undetectable in the animal body).This study was conducted to evaluate the competitive enzyme-linked immunosorbent assay (ELISA) system in order to detect the antibodies of przhevalskiana larvae in the goat sera. In order to prepare the larval antigen, 200 first instar larvae of przhevalskiana were collected from the subdermal region of the back and flanks of the infested goats in Khoramabad slaughterhouse, Khoramabad, Iran, from September 2017 to March 2018. Totally, 37 and 46 sera samples were taken from the infected and uninfected goats. The sensitivity and specificity at cut-off 3SD were obtained at 89.18% and 84.78%. Moreover, the measures of inter-and intra-assay coefficients of variability to express the precision or replicability of ELISA kit results were 5.33% and 2.81%, respectively.

Molecular and Serological Study of Neospora caninum Infection among Dogs and Foxes in Sanandaj, Kurdistan Province, Iran.

Neospora caninum, a protozoan parasite, causes abortions in cattle, as well as neurological disorders and reproductive problems in dogs. This study aimed to investigate the serological and the molecular prevalence of N.caninum among foxes and dogs using indirect fluorescent antibody test (IFAT) and nested-polymerase chain reaction (PCR). For this purpose, 288 and 95 both fecal and serum samples of dogs and foxes were collected, respectively, from around industrial and traditional dairy flocks in different parts of Sanandaj, Kurdistan Province, Iran, from 2013 to 2015. The sera were examined using IFAT, and fecal samples were microscopically assessed for detecting Neospora oocyst and by nested-PCR. The findings revealed that N.caninum seroprevalence were 4.86% and 4.21% in dogs and foxes, respectively. In addition, no Neospora oocysts were found microscopically and by PCR. Since this study is the first serological and molecular investigation of N.caninum among both dogs and foxes in Sanandaj, the findings of indicated that stray dogs is a main source of N.caninum infection in dairy farms in Sanandaj, Iran.

Molecular and Microscopic Detection of Theileria spp. among Cattle and Buffaloes in West Azarbaijan, Iran.

Bovine theileriosis is an important tick-borne disease caused by intraerythrocytic parasites from genus Theileria. This study sought to detect the theileriosis among cattle and buffaloes using molecular and microscopic tests in West Azerbaijan, Iran. For this purpose, 484 blood samples from 193 cattle and 291 buffaloes were collected during March to July 2014. The breed, gender, age, and habitat of these animals were recorded. These animals were native and apparently healthy, living in four different cities of the province. The blood films were stained with Giemsa’s for microscopic examinations. Direct cell semi-nested polymerase chain reaction (PCR) assay was performed to detect T.annulata DNA with Tbs-S/Tbs-A and To-S/Tbs-A primer pairs targeted to 18S ribosomal RNA gene for Theileria spp. and T.orientalis amplification, respectively. The molecular assays revealed that 36 cattle (18.65%) were infected, in which 15 cattle were infected by both T.annulata and T.orientalis. Out of 291 buffaloes, four samples (1.4%) were infected by Theileria genotypes, and two buffaloes (0.7%) were infected only by T.orientalis. The observational results of the gender, age, and habitat of the studied animals were similar to animals of the other parts of Iran. The present study indicated that T.orientalis may be prevalent in native cattle and buffaloes throughout the northern parts of Iran. This study assessed the infection of buffaloes with T.orientalis for the first time.

Detection of specific hydatid antigens and antibodies in serum and urine of experimentally infected sheep.

Immunodiagnostic confirmation of cystic human hydatidosis is frequently required before surgical intervention or of chemotherapy. However, it remains inadequate to detect specific antibodies or antigens in some confirmed cases of echinococcosis. This study was carried out to investigate the accuracy of three different immunodiagnostic tests for detection of specific circulating antigens or antibodies in the serum and urine of 13 experimentally infected sheep. For this purpose, Echinococcus granulosus were collected from small intestine of experimentally infected dogs, and 2000 taenid eggs were orally administered to each of the 13 sheep. There were six other sheep, which were kept as the control group. Biweekly serum and urine samples were collected from all the sheep for 4 months after infection. The sera were subjected to indirect hemagglutination test and the concentrated urine samples were subjected to coagglutination and counter immunoelectrophoresis tests. The results revealed that the sensitivity of these tests in detecting the hydatid antigens in the urine or antihydatid antibodies in the serum of the infected sheep reached their maximum in 12th and 13th week after infection; then it decreased in the following weeks. Examination of the non-infected sheep samples throughout the experiment showed that the aforesaid findings were specific only to the infected sheep. It seems that the appearance of specific hydatid antigen in urine and its antibodies in the serum were simultaneous. Although these tests are highly specific, false negative outcomes were encountered in their detection of cystic echinococcosis. In general, it seems rational to establish some series of diagnostic procedures in order to reveal antibodies and antigen of metacestode in serum and urine of the patients.

Seroprevalence of anti-Toxoplasma gondii antibodies in buffaloes in Khoozestan province, Iran.

The sera of 385 buffaloes of different age and sex, collected from the various areas of the Khoozestan province, Iran, were tested by the IFA test, from April 1995 to February 1996. The results indicated 8.8% (34 out of 385) buffaloes had anti-Toxoplasma gondii antibodies titered 1:16 and more. The infection rate of buffaloes under the age of a year, above a year female, and male were 10.8%, 4.7%, 12.1%, and 5.3%, respectively.

Progress in urinary schistosomiasis control measures in Iran.

During a ten-year field survey from 1980 to 1989, 1518 cases of Schistosoma haematobium were detected in Khoozestan province, the only area in south-west Iran where urinary schistosomiasis is prevalent. Most of the cases were detected by urine examination and a few by bladder biopsies. The incidence of infection was 0.653% in 1980, 0.021% in 1988 and 0.042% in 1989. Strict control measures, including mass chemotherapy and mollusciciding, were used to help reduce the prevalence of urinary schistosomiasis. The 20-29-year-old age group was the most infected; there were no sex differences.

Explanatum explanatum (Creplin, 1847) Fukui, 1929, in buffaloes in the Ahwaz area, southwest Iran.

Explanatum explanatum (Creplin, 1847) Fukui 1929, was found in three indigenous buffaloes in the west Ahwaz area, southwest Iran for the first time. The presence of this amphistome in bile ducts caused remarkable fibrosis of submucosa ad hyperplasia of the endothelial layer, such that villus-like structures and protuberances were formed in large bile ducts of the infected livers.

Prevalence of toxoplasmosis in humans and domestic animals in Ahwaz, capital of Khoozestan Province, south-west Iran.

From 1984 to 1988 sera from 1806 apparently healthy persons and 1096 patients suspected of toxoplasmosis in and around Ahwaz city, capital of Khoozestan province, Iran were examined by immunofluorescent antibody test (IFA). Prevalence of Toxoplasma antibodies at a titre of < or = 1:20 was found in 49.6% of healthy persons compared to 72.3% in suspected patients. The seropositivity rate was lowest in the new-born and children under 6 months of age and highest in persons 14-19 years old. Blood samples from 142 cattle, 138 sheep and 130 goats, collected from Ahwaz abattoir, were tested by the latex-agglutination test (LAT). The infection rates were 14.8, 13.8 and 13.1% respectively. The consumption of uncooked meat seems to be the most probable cause of the high infection rate of toxoplasmosis in the area.

Capillaria philippinensis infection in Khoozestan Province, Iran: case report.

Intestinal capillariasis was diagnosed in a 30-year-old fisherman from Malihan Village, Khoozestan Province, Iran. He reported a more than two month history of diarrhea, borborygmus, muscular atrophy, and debility. Eggs, larvae, and adults of Capillaria philippinensis were found in his feces. Treatment with mebendazole 400 mg per day for 25 days resulted in clinical and parasitological cure. The patient had reported eating a few small viable fish almost two months before clinical symptoms appeared. This is the first report of this infection from the Middle East.