Molecular prevalence and phylogenetic analysis of hemotropic Mycoplasma species in cats in different regions of Iran.

BACKGROUND: Hemotropic Mycoplasma species (hemoplasmas) cause hemolytic anemia in cats worldwide and are recognized as emerging zoonotic pathogens. There is no comprehensive study on the prevalence and species diversity of hemoplasmas in domestic cat populations in different regions in Iran. Thus, the aims of the present study were to provide data on the prevalence and molecular characterization of hemotropic Mycoplasma species in apparently healthy cats from six Iranian provinces with different climates. In addition, potential risk factors associated with hemoplasmosis in cats were assessed. RESULTS: Mycoplasma spp. DNA was detected in the blood of 56 / 361 cats (15.5%) using genus-specific PCR. Further examinations with species-specific PCR and Sanger sequencing showed that 38 cats (10.5%) tested positive for Candidatus Mycoplasma haemominutum (CMhm), 8 cats (2.2%) tested positive for Mycoplasma haemofelis (Mhf), and 2 cats (0.6%) tested positive for Candidatus Mycoplasma turicensis (CMt). Co-infection with CMhm, and Mhf was observed in 7 cats (1.9%). One cat (0.3%) showed mixed infection with CMhm, Mhf, and CMt. There were statistically significant relationships between Mycoplasma positivity and being female, living in shelter (cattery), and being over 3 years old (P < 0.05). No significant association was observed for the cat breed and sampling localities. CONCLUSIONS: Current study findings revealed that hemoplasma infections are common among Iran cat populations. Considering the impact of such emerging zoonotic pathogens on the One Health, routine screenings, increasing public awareness, effective control, and prophylactic strategies for minimizing infection in cats and subsequently in human are strongly recommended.

Molecular investigation of Leishmania in sandflies (Diptera: Psychodidae) and rodents (Mammalia: Rodentia) in Nahavand, west of Iran.

Cutaneous leishmaniasis caused by different species of Leishmania is transmitted by Phlebotominae sandflies. This disease remains a public health concern in Iran. Therefore, the present study aimed to examine Leishmania infection in sandflies and reservoir rodents in six rural regions of Nahavand, located in western Iran. From May to October 2022, sandflies and rodents were collected and identified at the species level. Additionally, rodents' skin lesions and earlobe specimens were collected separately for microscopic and molecular examination. All specimens were tested for Leishmania DNA by PCRs targeting the parasite's ITS-2 and 18S rRNA gene and positive were Sanger sequenced. A total of 3396 sandflies belonging to seven subgenera and 11 species, i.e., Phlebotomus papatasi (42.7%), P. major (20.6%), P. mascitti (0.3%), P. neglectus (0.2%), P. alexandri (0.2%), P. turanicus (0.3%), Sergentomyia murgabiensis (18.1%), S. dentata (10.5%), S. theodori (5.8%), S. antennata (1.1%), and S. pawlowski (0.1%) were identified. Based on the species population, 29 pools of sandflies were examined for the presence of Leishmania DNA using conventional PCR (cPCR), and individual DNAs were tested when positive. Leishmania major DNA was detected in two P. papatasi and Leishmania sp. in one P. major individual sandfly. This is the first report of Leishmania infection in sandflies from Hamadan province. The captured rodents (n = 61) belonged to four families and seven species, i.e., Arvicola amphibius (37.7%), Mus musculus (29.5%), Microtus socialis (13.1%), Apodemus sylvaticus (11.5%), Talpa davidiana (4.9%), Apodemus witherbyi (1.6%), and Rattus norvegicus (1.6%). Microscopic and molecular examinations of the rodent lesions and earlobes scored negative results. The presence of Leishmania in the Phlebotominae sandflies in Nahavand indicates a potential threat to humans and animals in the region. Regular monitoring and examination of the sandflies' population and timely diagnosis and treatment of new patients are strongly recommended.

Molecular identification, risk factor assessment, and phylogenetic analysis of tick-borne pathogens in symptomatic and asymptomatic cattle from South-Eastern Iran.

Tick-borne pathogens (TBPs) represent a substantial threat to cattle globally, exerting adverse impacts on production, health, and economic viability. This study delves into the prevalence and implications of TTBPs in cattle sourced from resource-limited smallholder livestock farms situated in southeastern Iran, proximate to Afghanistan and Pakistan. Blood and tick specimens were systematically collected from a cohort of 230 cattle, comprising 150 asymptomatic and 80 symptomatic individuals. Genomic DNA isolated from blood samples underwent rigorous examination for the presence of key TBPs, including Anaplasma marginale, A. phagocytophilum, A. bovis, A. centrale, Babesia bigemina, and Theileria annulata, utilizing multiple genetic markers. Nucleotide sequence analysis facilitated the reconstruction of phylogenetic relationships. The study also evaluated various potential risk factors, such as clinical status, gender, age, breed, tick infestation, and management practices, to elucidate their associations with TTBPs. Among the cattle cohort, a staggering 87.8% (202/230) tested positive for at least one pathogen. Prevalence statistics encompassed A. marginale (72.2%), T. annulata (68.3%), A. phagocytophilum/A. platys-like complex (66.1%), A. centrale (16.7%), B. bigemina (10.0%), and A. bovis (6.1%). Remarkably, mixed infections involving two, three, and four pathogens were detected in 23%, 52.1%, and 2.2% of animals, respectively. Notably, all asymptomatic cattle were positive for at least one TBP. Tick infestation was observed in 62.2% (143/230) of cattle, predominantly caused by Hyalomma anatolicum (82.5%), Rhipicephalus (Boophilus) annulatus (13.1%), and R. sanguineus sensu lato (4.4%). Risk factors linked to TBPs encompassed tick infestation, older age, and crossbred animals. Clinical presentations among symptomatic cattle encompassed fever, anemia, weight loss, anorexia, jaundice, and enlarged superficial lymph nodes. This study underscores the pivotal role of asymptomatic carriers in the propagation of TTBPs within endemic regions. Furthermore, it emphasizes the potential for the implementation of molecular diagnostics to unmask subclinical infections, thereby affording the opportunity for targeted interventions aimed at ameliorating the burden of TTBPs in resource-constrained smallholder dairy farms.

Neospora caninum and Toxoplasma gondii infections in one-humped camels (Camelus dromedarius) in central desert of Iran.

The protozoan parasite Neospora caninum infects carnivores as definitive and a wide range of mammals as intermediate hosts. This parasite is regarded as an important cause of abortion in cattle worldwide, causing significant economic losses. Although there is serological evidence of infection in Old World camelids, the significance of N. caninum in these animal species is still poorly understood. The aim of this study was to use molecular and histological methods to detect N. caninum in the blood and tissues of 100 slaughtered one-humped camels (Camelus dromedarius) in Iran. For this, genomic DNA was extracted from blood, brain, portal lymph node and liver of the camels, and nested-PCR assay followed by sequencing were performed. Besides, paraffin-embedded tissue sections were stained with hematoxylin and eosin (H&E) and studied microscopically. In addition, immunohistochemical staining for N. caninum was attempted on brain samples with positive PCR results. All animals were tested for antibodies against N. caninum and Toxoplasma gondii by whole tachyzoite-agglutination tests. N. caninum DNA was detected in blood, brain, and portal lymph node, but not in the liver of two (2%) camels. Histopathological examination revealed cysts resembling N. caninum in brain samples of one of these camels; however, immunohistochemical staining for N. caninum and T. gondii did not allow a morphological identification. IgG antibodies to N. caninum and T. gondii were detected in 36% and 35% of the camels, respectively. This study provides the first insight into direct detection of N. caninum in C. dromedarius in Iran. Further molecular studies on aborted fetuses, stillborn animals and cases of perinatal mortality are needed to understand the possible involvement of N. caninum in cases of reproductive failure. As the definitive hosts of N. caninum are domestic and wild canids, producers should be advised to monitor and limit exposure of their camelids to these species and their feces.

Hepatic Capillaria hepatica (Bancroft, 1893) infection in cat (Felis catus)-histopathological findings and first report from Iran.

Capillaria hepatica (syn. Calodium hepaticum) is a globally distributed nematode with a high affinity to the liver of a wide range of mammalian hosts, including humans. Documented reports of the nematode in cats and associated histopathology are rare. Here, we describe a case of C. hepatica infection in a 5-year-old male stray cat from Iran. At post-car accident necropsy, all body parts appeared normal except for the liver, in which a few yellowish-white granulomatous nodules were observed through the capsule and in the organ. Histopathological examination of the tissue revealed a large number of clustered parasite eggs in the parenchyma. The barrel-shaped, un-embryonated eggs (55.19 × 28.37 µm), with inconspicuous caps at both ends, were covered with striated shells. The presence of ova in the liver tissue had resulted in the development of hepatic inflammation with hepatocellular necrosis associated with the development of multifocal granulomas. As predators of small rodents, the cats might have a significant role in the epidemiology of C. hepatica. Infection of hosts through ingestion of embryonated eggs in contaminated water, food, or soil is of major importance in the epidemiology of C. hepatica. Since the rare reports of feline infection have come mainly from accidental detection of the parasite, any hepatic disease presenting difficulties to find an etiological agent may virtually be associated with the infection with this little-known nematode.

Vector-borne pathogens in dogs of different regions of Iran and Pakistan.

Canine vector-borne diseases (CVBDs) are highly prevalent in tropical and subtropical countries, mainly due to favorable climate conditions and reduced adoption of preventive measures. This study aimed to provide a comprehensive overview on the prevalence of CVBDs in Iran and Pakistan where limited data are available. Blood samples were collected from 403 dogs from six provinces in Iran and Pakistan to assess the presence of pathogen DNA (i.e., Anaplasma spp., Coxiella burnetii, Ehrlichia spp., Rickettsia spp., Babesia spp., Hepatozoon spp., filarioids, and Leishmania spp.). Sera were also screened by an immunofluorescence antibody test for the detection of antibodies against Leishmania infantum. In total, 46.9% of dogs scored positive to Hepatozoon canis being the most frequently detected (41.4%), followed by Anaplasma platys (6.4%), Ehrlichia canis (3.4%), Rickettsia spp. (2.2%), Babesia vogeli (1.0%), and L. infantum (0.3%). A seroprevalence of 9.6% to anti-L. infantum IgG was also recorded. Data reported herein demonstrate that dogs from Iran and Pakistan are at a high risk of CVBDs, particularly of canine hepatozoonosis. Effective control strategies are advocated for minimizing the risk of infection in animals and humans, also in consideration of the zoonotic potential of some pathogens detected.

Molecular Detection of Zoonotic Pathogens in the Blood and Tissues of Camels (Camelus dromedarius) in Central Desert of Iran.

Dromedary camels (Camelus dromedarius) play a major economic role in many countries in Africa and Asia. Although they are resistant to harsh environmental conditions, they are susceptible to a wide range of zoonotic agents. This study aimed to provide an overview on the prevalence of selected zoonotic pathogens in blood and tissues of camels in central Iran. Blood, liver, portal lymph node, and brain were collected from 100 apparently healthy camels at a slaughterhouse in Qom city to assess the presence of DNA of Brucella spp., Trypanosoma spp., Coxiellaburnetii, and Bartonella spp. PCR products were sequenced bidirectionally and phylogenetic analyses were performed. Eleven percent of camels tested positive for Brucellaabortus (3%) and Trypanosomaevansi (8%). Coxiellaburnetii and Bartonella spp. DNA was not detected. Our data demonstrate that camels from Iran contribute to the epidemiology of some zoonotic pathogens. Performing proper control strategies, such as vaccination of camels and humans in contact with them, test-and-slaughter policy, and education of the general population is necessary for minimizing the risk of zoonotic infection.

Anaplasma ovis infection in sheep from Iran: molecular prevalence, associated risk factors, and spatial clustering.

The aim of this cross-sectional study was to determine the molecular prevalence and associated risk factors in sheep populations of Iran. To this end, between March 2017 and February 2018 jugular vein blood samples were collected from 1842 apparently healthy sheep from 327 herds in nine provinces in four ecological zones of Iran. A specific nested-PCR targeting the msp4 gene of A. ovis was employed. Fourteen variables were subjected to logistic regression analyses (univariate and multivariate) to specify the potential risk factors for infection. Statistically significant variables in univariate analyses (P ≤ 0.20) were assessed by multivariable logistic regression to control the confounding factors. Anaplasma ovis DNA was detected in 51.1% of herds (167/327) and 28.3% of animals (521/1842). Among geographical zones, herd and animal prevalence was highest in the Persian-Gulf zone (P < 0.001), and among provinces, Lorestan (in west) and Khuzestan (in south-west) had the highest prevalence (P < 0.001). Analysis of factors associated with A. ovis infection revealed that distance from other farms (OR = 2.52, P < 0.001), presence of other animal species in the farm (OR = 2.03, P = 0.046), season (OR = 1.40, P = 0.005), breed (OR = 3.762, P < 0.001), and age of sheep (OR = 1.20, P = 0.049) are potential risks in Iran. The spatial scan statistic in SaTScan recognized two high risks clusters for A. ovis infection in central (Semnan province) and the Persian-Gulf (Khuzestan province) zones amongst the study areas (P < 0.001). Sequence and phylogenetic analysis of the msp4 gene confirmed the detection of A. ovis. This research is the largest study focusing on ovine anaplasmosis in Iran and shows that infected sheep are present in all geographic zones, bioclimatic areas, and provinces.

Molecular identification and phylogenetic analysis of Dipetalonema evansi (LEWIS, 1882) in camels (Camelus dromedarius) of Iran.

Despite the economic importance of camels, the parasites that affect them have not received adequate attention so far and molecular studies are scarce compared to other livestock. In this study, we characterized peripheral blood microfilariae in 200 healthy one-humped camels (Camelus dromedarius) from south-east Iran by microscopy and molecular tools to receive a more detailed insight into prevalence and species that affect them. Moreover, adult specimens of the filarial nematode Dipetalonema evansi were collected from the carcass of an infected animal. Microscopic examination was performed on Giemsa-stained blood smears, and blood was also spotted on Whatman FTA(®) cards for DNA analysis. Genomic DNA was extracted, and PCR was carried out for the detection of filaroid helminths, followed by sequence analysis of positive samples. Four samples were positive for microfilariae by microscopy, while 16 animals (8 %) were positive by PCR. Sequence analysis revealed D. evansi in all cases. Phylogenetic analysis of a cytochrome C oxidase subunit I (COI) sequence of filaroid nematodes showed that most species in a single genus cluster in the same clade; however, D. evansi and D. gracile are not monophyletic and branch rather at the base of the tree. Further studies on the life cycle of D. evansi, specifically the identification of intermediate host(s), have become feasible with the provision of the first specific COI sequences in this study.

Identification and characterization of a cathepsin L-like cysteine protease from Rhipicephalus (Boophilus) annulatus.

The tick Rhipicephalus (Boophilus) annulatus is one of the most important ectoparasites of bovines and is responsible for the transmission of different pathogens such as Babesia and Anaplasma. Cysteine proteases are involved in several host-tick interactions including invasion of host tissues, immune evasion, pathogen transmission, embryogenesis and blood digestion. In this study, the gene encoding R. annulatus cathepsin L-like enzyme (RaCL1) was cloned into pTZ57R/T vector, sequenced and analyzed using bioinformatics approaches. The nucleotide length of RaCL1 was 999 bp. Bioinformatics analysis showed 332 amino acids with an approximate molecular weight of 36.3 kDa which contained a signal peptide sequence (18 amino acids), pro-region (97 amino acids) and mature enzyme (217 amino acids). Multiple sequence alignment of the RaCL1 revealed high similarity to cathepsin L-like cysteine proteases from other tick species such as Rhipicephalus (Boophilus) microplus and Amblyomma variegatum. Based on bioinformatics analyses, results of this work suggest that RaCL1 can be a suitable candidate for the development of vaccine against R. annulatus.

Molecular detection and identification of hemotropic Mycoplasma species in dogs and their ectoparasites in Iran.

Hemotropic Mycoplasma species are vector-borne bacteria that attach and grow on the surface of erythrocytes in various mammals, yet reports of canine hemoplasmosis in Iran are scarce. The aim of this study was molecular detection and identification of hemoplasmas in the blood of dogs (n = 370) from five provinces of Iran and ectoparasites infesting them including Ctenocephalides canis and Pulex irritans fleas, Rhipicephalus sanguineus sensu lato ticks, Heterodoxus spiniger lice and Hippobosca longipennis keds. Hemotropic Mycoplasma spp. pathogens were detected using genus-specific conventional PCRs, and subsequently identified using species-specific PCRs for Mycoplasma haemocanis (Mhc), and Candidatus Mycoplasma haematoparvum (CMhp). Sanger sequencing was then performed to confirm the species. Correlation of infection and risk factors (geographical area, keeping condition, body condition, sex, age, ectoparasite infestation) were analyzed. In total, 210 dogs (56.7%) were tested PCR-positive for hemotropic Mycoplasma spp. Species-specific PCR and sequencing revealed infection with Mhc in 17.8%, with CMhp in 7.02% and co-infection in 31.9% of dogs. Flea infestation, poor body condition, and being older than 3-years-old correlated with hemoplasmosis. In ectoparasites, DNA of hemoplasmas were detected only in fleas i.e. Mhc in P. irritans, CMhp in P. irritans and C. canis, and co-infection in C. canis. To our knowledge, this is the first large-scale molecular epidemiology study of canine hemoplasmosis in Iran. Considering the high prevalence of canine hemoplasmosis all over the country including potentially zoonotic CMhp, effective ectoparasite control strategies, regular examination of dogs, successful chemoprophylaxis and public awareness strategies are advocated.

First record of nasopharyngeal myiasis caused by Cephalopina titillator (Clark, 1816) in camel (Camelus dromedarius Linnaeus, 1758) in Uzbekistan.

Nasopharyngeal myiasis caused by the camel nasal bot, Cephalopina titillator, is very common in old world camelids and is usually found at necropsy or during meat inspection. Herein we report massive infection with C. titillator in a 9 years old female one-humped camel slaughtered on February 18, 2024 in the village of Kizil Uy, Nukus District, Republic of Karakalpakstan, northwestern Uzbekistan. A total of 69 larvae: 20 first stage larva (28.9%), 31  second stage larva (44.9%), and 18 third stage larva (26.0%) were detected in the nasal passages and pharynx of the camel. Morphological and morphometrical characters of all larval stages are illustrated and detailed in this article. To our knowledge this is the first record of camel nasal bot infestation in Uzbekistan. Future epidemiological studies are needed to shed light on the prevalence, seasonal fluctuation, clinical impact and economic burden of nasopharyngeal myiasis in dromedary camels of the country.

Impaired efficacy of fenbendazole and ivermectin against intestinal nematodes in adult horses in Iran.

This study aimed to evaluate the efficacy of fenbendazole and ivermectin on strongyles and Parascaris sp. infecting adult riding horses in three regions with different climates. During 2021 and 2022 fecal specimens were randomly collected from 483 horses older than three years in 31 equestrian clubs in Hamedan (n = 217), Yazd (n = 146) and Tabriz cities (n = 120). Eggs were counted by McMaster technique, and the strongyle larvae were identified using coproculture, PCR and sequencing. Horses with strongyles and Parascaris egg counts ≥150 were enrolled in fecal egg count reduction (FECR) examination following treatment with ivermectin or fenbendazole. In total, 26.5% of examined horses were positive with at least one parasite. Infection rates varied in three cities i.e., 25.8% in Hamedan, 28.8% in Yazd, 25% in Tabriz. Fifty-seven horses had FECR measured. FECR below <90% was observed for IVM-strongyle in two horses in Tabriz, for FBZ-strongyle in two horses in Tabriz and two horses in Hamedan, for IVM-Parascaris in one horse in all three cities, and for FBZ-Parascaris in one horse in Yazd. Furthermore, FECR 90-100% was observed in IVM-Parascaris and FBZ -Parascaris groups in Tabriz. Data herein presented demonstrate different degrees of resistance of strongyles and Parascaris infecting horses in Iran against both ivermectin and fenbendazole. Since non-principled use of anthelminthics is common among horse owners, urgency of test-and-treatment strategy should be educated and implemented by policy-making organizations. Evaluating efficacy of different anthelminthics and choosing the most effective treatment in each region is suggested.

Molecular identification of hemoparasites in ixodid ticks in Iran.

A total of 386 ticks were processed in order to investigate the occurrence of selected tick transmitted pathogens (i.e., Theileria, Babesia, Hepatozoon and Cytauxzoon) in ixodid ticks in six provinces of Iran (East Azerbaijan, Gilan, Kermanshah, Khuzestan, Sistan & Baluchestan and Tehran). Ticks identified as Dermacentor marginatus, Hyalomma aegyptium, Hy. anatolicum, Hy. asiaticum, Hy. marginatum, Ixodes ricinus, Rhipicephalus annulatus and R. sanguineus sensu lato were collected from sheep and cattle. Conventional PCR and sequencing results revealed DNA of Theileria ovis in three R. sanguineus sensu lato pools and one D. marginatus pool from sheep in Kermanshah and East Azerbaijan, T. annulata in one Hy. asiaticum pool from cattle in Kermanshah, and He. canis in an individual female Hy. aegyptium in Kermanshah. Data herein indicate the role of R. sanguineus complex and D. marginatus in the epidemiology of ovine theileriosis in western and northwestern Iran. Potential role of Hyalomma aegyptium in the transmission of He. canis is discussed. Considering non-principled movement of livestock across the country and increasing reports about the resistance of ticks to common acaricides, test-and-treatment of infected livestock, vaccination of the livestock against endemic tick-borne pathogens, and the use of non-chemical tick management strategies are recommended.

Wildlife parasitology: sample collection and processing, diagnostic constraints, and methodological challenges in terrestrial carnivores.

Wild terrestrial carnivores play a crucial role as reservoir, maintenance, and spillover hosts for a wide parasite variety. They may harbor, shed, and transmit zoonotic parasites and parasites of veterinary importance for domestic hosts. Although wild carnivores are globally distributed and comprise many different species, some living in close proximity to human settlements, only a few studies have investigated parasites of wild terrestrial carnivores using non-specific techniques. Access to samples of wild carnivores may be challenging as some species are protected, and others are secretive, possibly explaining the data paucity. Considering the importance of wild carnivores' health and ecological role, combined with the lack of specific diagnostic methodologies, this review aims to offer an overview of the diagnostic methods for parasite investigation in wild terrestrial carnivores, providing the precise techniques for collection and analysis of fecal, blood, and tissue samples, the environmental impact on said samples, and the limitations researchers currently face in analyzing samples of wild terrestrial carnivores. In addition, this paper offers some crucial information on how different environmental factors affect parasite detection postmortem and how insects can be used to estimate the time of death with a specific highlight on insect larvae. The paper contains a literature review of available procedures and emphasizes the need for diagnostic method standardization in wild terrestrial carnivores.

Gastrointestinal parasites of cats in the Middle East (2000-2023): A literature review.

BACKGROUND/OBJECTIVE: Gastrointestinal parasites in stray cats are of significant clinical and zoonotic importance and pose a potential threat to public health. Hence, the current review has practical implications as it aims to present the diversity and the prevalence of cat parasites in Middle Eastern countries. METHODS: The research process was rigorous and thorough, spanning from 2000 to 2023. Data collection and analysis were conducted utilizing multiple international and national databases, including Google Scholar, Scopus, PubMed, and Irandoc. This meticulous approach ensures the reliability and validity of the findings, providing a solid foundation for further research and preventive measures. RESULTS: A total of 159 articles were collected and comprehensively reviewed. Among these, 85 articles were from Iran, 25 from Türkiye, 20 from Iraq, 11 from Egypt, five from Qatar, three from Cyprus, three from Kuwait, and one each from the UAE, Saudi Arabia, Syria, Lebanon, Jordan, Israel, and Palestine. Toxocara cati was the most prevalent nematode, exhibiting the highest prevalence in Iran, ranging from 8% to 90%, followed by Egypt, with a range of 8.23% to 58.7%. In comparison, the lowest prevalence was recorded in Qatar at 8%. Hydatigera taeniaeformis, a common cestode in the feline, showed the highest prevalence in Qatar, with a range of 73.6% to 75.8%, while the lowest was in Kuwait at 0.8%. In Iran, the prevalence ranged from 0.9% to 64.7%. Toxoplasma gondii, a cosmopolitan protozoan, showcased the highest prevalence in Lebanon at 78.1% and the lowest in the UAE at 0.8%. In Iran, the prevalence ranged between 1.2% and 90%. Additionally, noteworthy patterns regarding the distribution and potential risk factors associated with these parasites were observed. While gender variations were not significant, environmental conditions such as high humidity and absence of sunlight exposure, along with the age and behavior of cats, were among the risk factors associated with the prevalence of parasites. CONCLUSION: The prevalence of parasites among stray cats in Iran and other Middle Eastern countries remains notably high. This emphasizes the urgent need for a series of control and preventive measures. Implementing effective anti-parasitic strategies and emphasizing the development of multiple diagnostic methods are essential.

Eimeria leuckarti in equid coprolites from the Sassanid Era (2nd-6th century CE) excavated in Chehrabad Salt Mine archaeological site, Iran.

OBJECTIVE: This study reports coccidian oocysts in an equid coprolite dated to the Sassanid Empire (2nd-6th century CE) recovered in Chehrabad Salt Mine archaeological site, Iran. METHODS: Between 2015 and 2017, an archaeoparasitological investigation led to the discovery of an equid coprolite in the Chehrabad Salt Mine archeological site, (Douzlakh), western Iran. Samples were rehydrated using trisodium phosphate solution and were examined by light microscopy. RESULTS: Seven oocysts of Eimeria leuckarti (Flesch, 1883) were identified; they were in various stages of sporulation. CONCLUSION: This is the first report of ancient coccidian oocysts from equids. The importance of this observation is discussed, and current knowledge of eimeriid oocysts at archaeological sites is reviewed. SIGNIFICANCE: The observations of E. leuckarti increases current knowledge of parasite biodiversity in ancient Iran when it rested along the Silk Road, a network of trade routes connecting the East and West that was central to economic, cultural, political, and religious interactions between these regions, and to livestock movement that could contribute to the transmission of the parasites from/to other regions. LIMITATIONS: The contextual information about animal species present in and around the Salt Mine during its working periods, including Achaemenid dynasty (6th to 4th century BCE) and Sassanid era (2nd to 6th century CE), is very limited and does not allow secure conclusions regarding the host origin of the coprolites. SUGGESTIONS FOR FURTHER RESEARCH: Application of molecular biology tools to identify the correct host origin of the coprolites and to detect more parasite species is advocated.

Biliary parascarosis in a foal.

Migration of Ascaris lumbricoides through the papilla of Vater in humans, and entry into the biliary tree, is well-recognised. Ascaris suum and Toxocara vitulorum have been recovered from the liver of swine and buffalo. We necropsied a Persian Kurdish filly at age 6 months, weighing ∼100 kg. Death evidently was caused by oleander (Nerium oleander) intoxication. An 8-cm adult male Parascaris was found at the lobar-left hepatic bile duct junction. We suggest that the nematode entered anteriorly into the hepatic tree, via the duodenum, major duodenal papilla, bile duct, left hepatic duct and finally the lobar duct. Considering the brief 4-h elapsed time between death and necropsy, and the 18-cm distance from the major duodenal papilla to the location of the parasite, we conclude that entry into the biliary tree likely occurred ante-mortem. We advise consideration of Parascaris infection in differential diagnosis of equine hepatic and pancreatic dysfunction.

Lungworms of Non-Ruminant Terrestrial Mammals and Humans in Iran.

With over 300 terrestrial and aquatic mammalian species, Iran is considered a country with an ample mastofauna. Although many studies have assessed the distribution of gastrointestinal helminth parasites in animals and humans in Iran, lungworms have not received adequate attention. Following a previous article in which we reviewed the diversity and prevalence of lungworm infections in pastoral and wild ruminants of Iran, this report compiles the available scientific information about the occurrence of lungworms in non-ruminant mammals and humans from 1980 to 2022 to provide insights into the epidemiology of these infections. International and national scientific databases were searched, and twenty-six articles in peer-reviewed journals, one conference paper, and one D.V.M. thesis were included in the study. In total, 10 species belonging to seven genera, including Dictyocaulus, Deraiophoronema, Protostrongylus, Crenosoma, Eucoleus, Aelurostrongylus, and Metastrongylus, were reported in the respiratory tract or feces of humans, domestic animals (i.e., camels, equids, dogs, and cats), and wildlife species (i.e., hedgehogs, wild boars, and hares). Most of the studies (22/28) were performed using post-mortem examinations. The overall prevalence of respiratory nematode infection varied according to animal species in camels (14.83%), equids (13.31%), dogs (5%), wild boars (45.66%), hedgehogs (42.57%), and hares (1.6%). In addition, pulmonary capillariasis caused by Eucoleus aerophilus was reported in a 9 year old child. The prevalence of lungworm species in domestic camels, equids, and dogs, combined with a lack of labeled anthelmintic products, supports the need to improve our understanding of these important nematode parasites and inform the development of sustainable control strategies. From a zoo and wildlife medicine point of view, there is a shortage of information about the presence and prevalence of lungworm infections in the majority of mammalian species, pending epidemiological studies that integrate classical parasitology and molecular methods.