Linguatula serrata is parasitic zoonoses with cosmopolitan distribution. The adults are seen in the nasal and airways of canines as the definitive host while larval stages more in mesenteric lymph nodes ruminants as intermediate hosts. Prevalence of the parasite in ruminants of different areas of Iran and the world has been done mostly based on slaughterhouse investigations. So the goal of the study was to determine seroprevalence of linguatula serrata infection in goats in Ilam and Ahvaz (slaughtered animals) by indirect enzyme-linked immunosorbent assay (ELISA) to provide epidemiological information of the parasite for control measurements. In the study Excreatory-secretory antigen were prepared by incubation the larvae in RPMI media. Up to1172 goat sera of five regions of Ilam and Ahvaz were examined by ELISA. Data were analyzed by using statistical software (SPSS). There was significant difference in prevalence of the infection among five different areas of the Ilam province. The infection in goat in the city of Dehloran(South) 19/2%, in the city of Ivan (North) 63/4%, in city of Chardavol (East) 16/9%, in city of Mehran (West) 9/9% and in the center ( Ilam) 65/8%. Also the infection in goat in the city of Ahvaz 38/5%. The infection rate of the parasite in male ( Ilam) 32/2 % and females 24/6% was significantly associated with sex of the animals, while infection rate of the parasite in males 36/7% and in females 39/7% from Ahvaz was not significantly associated with sex of the animals. Seroprevalence of the infection in Ahvaz was increased significantly with the increase of ages. While there was not significant difference in prevalence of the infection of ages in Ilam's animals. There was not significant difference in prevalence among goat herds with dogs (26/7%) and those without dogs (30/8%). The results shows wasn’t significant in prevalence between groups (resident herds 29/1% and nomadic herds 26/9%). There was statistical significant difference between type of livestock and the infection. There was significant difference in prevalence among goat herds with have drugs (22/67%) and those without drugs (32/97%). Seroprevalence of the infection in summer (29/9%), in autumn (31/7%) and in winter (23/1%) were not sinificant difference between the seasons. Considering of the high linguatula serrata infection in goats of the areas, control measurements is very necessary.

<p>Linguatula serrata is one of the parasites with zoonotic importance in dogs. Herbivorous intermediate hosts such as sheep have a consequential role to survive and dispersal of the parasite. Aim of the study was conducted to determine prevalence of the infection in sheep from Ilam province and Ahvaz. In the study during 8 months from April to November 2015, 804 lymph node samples were taken from slaughtered sheep of Ilam province (five different geographical area) and Ahvaz (canter of Khuzestan province). Each sample (about 40 gram) the samples (of each animal about 40 grams) with registered separately region, gender, age were transferred to the laboratory. Lymph node status with regard to color (natural, hemorrhagic (red), black) or of consistency (normal, soft, hard) were determined. Positive samples were detected by collecting of Linguatula serrata nymphs. Nymphs collected from each sample were counted and transferred to Ethanol 70% then measured. A total of 619 samples of sheep from Ilam, 99 samples (16%) were infected with Linguatula serrata nymphs. The highest prevalence in the southern (19%) and the lowest (11.7%) in the eastern region of the province was determined. Statistically significant difference between the prevalence of the infection was no observed in sheep of different regions of the province (P&lt;0.05). Nymphal prevalence rate in female with 25.6% (52 out of 203) was revealed more than in male with 11.3% (47 out of 416). Relationship between sex and infection rate was statistically significant (P&lt;0.05). Based on age groups, the highest infection was related to 3&ge; year's groups with 23.4% and the lowest with 11.3% in under 1 year's groups. The relationship between age and the infection statistically at P&lt;0.05 was significant. Classification of the prevalence based on color of lymph nodes, most of the infection rates were seen in the black (dark) lymph nodes with 45.6% compared to red lymph nodes with 7.1%. Based on consistency of the lymph nodes, more of the infection rates were seen in the soft lymph nodes with 30.7% compared to normal lymph nodes with 7.6%. Relationship between color, consistency of lymph nodes and infection rate was statistically significant (P&lt;0.05). Of the total lymph nodes sample of 185 sheep in Ahvaz, 13 (7%) were infected with Linguatula serrata. Significant difference was seen between the prevalence of Linguatula serrata infection in sheep of Ilam province (16%) with prevalence in Ahwaz (7 %) at (P&lt;0.05). Size length ranges of the nymphs (fixed in 70% ethanol) were 2-12.3 mm with an average 5.64 mm. Due to the state of infection in sheep of the areas mentioned, control measures to reduce infection in the definitive hosts are necessary.</p>

Linguatula serrata is a zoonosis with world-wide distribution. Dogs and other carnivores are its main hosts and herbivores and some rodents are its intermediate hosts. It does not have any clinical symptoms in goat and other intermediate hosts and diagnosis and the prevalence of this parasite studies are mostly based on slaughterhouse and necropsy observations or histopathology tests. Therefore, the purpose of the present study was the evaluation of indirect haemaglutination test for serological detection of Linguatula serrata infection in goats using the parasite nymphal excretory-secretory and somatic antigens. The somatic antigen was made of collected lymph from mesenteric lymph nodes of goats and homogenizing them in25 lymph ml PBS- RPMI-1640 and the excretory-secretory antigen was made by growing 25 lymph ml PBS- RPMI-1640. An initial evaluation to determine the best antigen from among excretory-secretory, added somatic (with DTT or PMSF), and simple antigens was made. The obtained results indicated that simple antigens especially its excretory-secretory antigens had the best results and were better than the added ones. To evaluate the indirect hemagglutination, 40 positive serum samples from infected goats with parasite lymph and 40 negative serum samples of under 3 month-old goats in five different concentrations from 1/2 to 1/32 were used and sensitivity, specificity, positive predictive value, and negative predictive value with somatic antigens were estimated 7/5%, 97/5%, 75%, 51%, respectively, and for excretory-secretory antigens 83%, 85%, 86%, 81%, respectively. The results of serum prevalence of infection of Linguatula serrata in the goats slaughtered in Ahvaz with indirect hemagglutination test and excretory-secretory parasite antigen showed that 158 samples (57%) out of 281 examined serums were positive. Thus, indirect hemagglutination test using excretory-secretory antigens of Linguatula serrata nymphs can be evaluated to monitor pollution in goats, especially in epidemiologic studies. Considering the amount of infected goats in the region, control and prevention ways are necessary.

Oestrus ovis (sheep nasal bot fly) is an important parasite of domesticated small ruminants. Several cases of respiratory and ophtalmomyiasis manifestations of human myiasis caused by O.ovis larvae have been observed in Iran and some countries. Aim of this study was to evaluate Oestrus ovis larval excretory-secretory (ES) and somatic (S) antigens for immunodiagnosis of the infection in goats by counterimmunoelectrophoresis (CIE) and indirect haemaglutination (IHA). Third and second larval stages of Oestrus ovis were collected from cut horns of goats after tagging and blood sampling in Behbahan slaughterhouse. The collected larvae (L2 and L3) were washed several times with phosphate buffered saline (PBS) supplemented by antibiotics. For preparing of somatic antigens, larval stage (L2 and L3) were homogenized separately with PBS and dithiothreitol (DTT) by sonicator and centrifuged prior to collect supernatants for further steps. Excretory-secretory antigens (ES) of larval stages (L2 and L3) were prepared in cell culture flask containing RPMI media with antibiotic supplemented with 0.5% Co2, incubated for 24-48 hours. The media were centrifuged, after collecting the supernatants, filtered them, determined protein concentration according to Bradford assay. In the present study CIE and IHA methods were evaluated with 30 positive sera (from infected goats) and 30 negative sera (from indoor kids) to detection of Oestrus ovis infection in goats. Results of evaluation of the tests revealed the sensitivity and specificity rates of IHA test with ES antigens of second stage larvae, 91%, 85% and with somatic antigens 82% and 86% respectively. The sensitivity and specificity rates of IHA test were 86%, 100% for ES antigens and 23% and 92% respectively for somatic antigens of third stage larvae. Results of CIE with the antigens in comparison of IHA revealed that the CIE test were not of immunodiagnostic value for Oestrus ovis infection in live goats since sensitivity of CIE with ES antigens of L2 and L3 were 15.79% and 12% respectively. 206 sera samples from goats of the area (Behbahan) were evaluated for serodiagnosis of Oestrus ovis infection by IHA using ES antigens of L2 and L3. Infection rates in the animals were determined 59.7% and 43.2% with each antigens separately and 36.4% with both larval antigens. Results of the present study revealed that IHA rapid, inexpensive method with Oestrus ovis ES larval antigens can be used to detect the infection, especially in big herd goats for treatment, control and decrease of economic loss in the animals consequently reduction of the myiasis in humans.

Linguatula serrata causes a common disease (linguatulosis) in humans and some animals. In life cycle of the parasite, definitive hosts are some carnivores (dogs). Ruminants, certain mammals, and human are intermediate and dead end hosts respectively. Considering to role of ruminants to survive and transmission of the parasite to definitive hosts and human, the goal of the study was to determine seroprevalence of Linguatula serrata nymph infection in sheep from Kermanshah province by indiret enzyme-linked immunosorbent assay (ELISA) . Blood samples (n= 450) were taken from sheep of five different area or cities of Kermanshah province. The collected sera were tested by ELISA using excretory-secretory antigens of L. serrata nymph and anti IgG sheep conjugate. Data were classified according to sex of the animals, area, ages (1-<2, 2-<3, 3-≥4 years) and then analyzed using statistical software (SPSS). Seroprevalence of L. serrata infection was out of 125 (27.8%) in sheep from Kermanshah province. There was significant difference in prevalence of the infection among five different area of the province (P<0.05). The infection rate in sheep from northern area (cities) (Oramanat with mean of 45.3%) was high and those from southern area (Eslam Abade Gharb with 9.5%) low, there was significant difference statistically in prevalence among the areas. Of 225 males and 225 females, out of 68 (30.2%) and 57 (25.3%) were infected. Infection with the parasite was not significantly associated with sex of the animals (p>0.05). Seroprevalence was increased significanty with the increase of ages (Lower infection (9.3%) in (1-<2) and higher (46.9%) in (3-≥4) years old of the animals (P<0.05). This is the first report in assessment of L. serrata infection in sheep by ELISA. Prevalence of Linguatula serrata infection in the sheep of Kermanshah is relatively high, it must be consider to all programs for control the infection in major definitive and intermediate hosts.

Linguatula serrata causes a common disease (linguatulosis) in humans and some animals. In life cycle of the parasite, definitive hosts are some carnivores (dogs). Ruminants, certain mammals, and human are intermediate and dead end hosts respectively. Considering to role of ruminants to survive and transmission of the parasite to definitive hosts and human, the goal of the study was to determine seroprevalence of Linguatula serrata nymph infection in sheep from Ilam province by indiret enzyme-linked immunosorbent assay (ELISA) and relationship between presence or absence of dog in the sheep herds and the infection. Blood samples (n= 589) were taken in three seasons (spring, summer and autumn) from sheep of five different area or cities of Ilam province. The collected sera were tested by ELISA using excretory-secretory antigens of L. serrata nymph and anti IgG sheep conjugate. Data were classified according to sex of the animals, area, ages (1-<2, 2-<3, 3-≥4 years) and sheep herds with or without dogs, and then analyzed using statistical software (SPSS). Seroprevalence of L. serrata infection was out of 192 (32.1%) in sheep from Ilam province. There was significant difference in prevalence of the infection among five different area of the province (P<0.001). The infection rate in sheep from southern area (cities) (Dehloran and Abdanan with mean of 44.95%) was high and those from northern area (Ilam and sirvan with 23.15%) low, there was significant difference statistically in prevalence among the areas. Of 298 males and 300 females, out of 97 (32.5%) and 95(31.7%) were infected. Infection with the parasite was not significantly associated with sex of the animals (p>0.05). Seroprevalence was increased significanty with the increase of ages (Lower infection (14.5%) in (1-<2) and higher (41.4%) in (3-≥4) years old of the animals (P<0.05). There was significant difference in prevalence among sheep herds with dogs (59.5%) and those without dogs (22.3%) (P<0.001). This is the first report in assessment of L. serrata infection in sheep by ELISA. The highest Seroprevalence of the infection in spring (43.6%) and lower of them in summer (18.2%) were significanty different (P<0.05). Prevalence of Linguatula serrata infection in the sheep of Ilam is relatively high, it must be consider to all programs for control the infection in major definitive and intermediate hosts.

Linguatula serrata, is a cosmopolitan zoonotic parasite. Adult of L. serrrata parasitize the nasopharynx of canids (Final hosts). The larval form occurs in the mesenteric lymph nodes of herbivores (Intermediate hosts) including sheep. Infected vegetables, fruits, and water resources with eggs of the mature parasite excreted via carnivores’(especially stray dogs) nasopharyngeal secretions or feces is the main source of infecting human beings. However, consumption of infected improperly cooked viscera of the intermediate hosts including sheep, goats, cattle, or other herbivores containing the larval stages of this parasite is the other potential source of infection of human beings. Goal of this study was the isolation and identification of excretory secretory and somatic antigens from L. serrata by SDS-PAGE and immunoblotting.
The parasites were collected and washed by PBS supplemented by antibiotic for several times. For preparing somatic antigens, parasites were sonicated and centrifuged prior to collect supernatant. For preparing excretory secretory antigens viable parasites were transferred to the sterile medium. The samples were centrifuged and supernatants were collected. In order to prepare polyclonal antibody, two rabbits were immunized by L. serrata antigens. The sera of infected sheep with different infection degrees were collected too. Somatic and excretory-secretory proteins were isolated with sodium dodecyl sulphate (SDS-PAGE) and stained with coomassie blue. Immunogenic properties of the resulting proteins were determined using immunoblotting. The total extract of somatic antigens analyzed by SDS-PAGE revealed 18 proteins that bands of 25, 28, 36, 45, 48, 67, 75, 91, 100 and bond above 180 KDa were immune dominant in infected sheep. In excretory-secretory antigens 9 bands of protein were detected that bands of 38, 58 and above 180 KDa were immune dominant. Probably the most immunogenic protein band in infection with somatic and excretory secretory antigenis in sheep was a bove 180 KDa that can be used for further evaluations in immunodiagnostic and protection studies in sheep and other animals.