In vitro feeding of Hyalomma excavatum and Hyalomma marginatum tick species.

The rearing of ticks is an important technique for studies aiming to elucidate the course and pathogenesis of tick-borne diseases (TBDs). TBDs caused by protozoans (Theileria, Babesia) and bacteria (Anaplasma/Ehrlichia) impose a serious constraint upon livestock health and production in tropical and sub-tropical regions where the distributions of host, pathogen, and vector overlap. This study focuses on Hyalomma marginatum, one of the most important Hyalomma species in the Mediterranean region, being a vector of the virus that causes Crimean-Congo haemorrhagic fever in humans, together with H. excavatum, a vector of Theileria annulata, an important protozoan of cattle. The adaptation of ticks to feeding on artificial membranes allows the creation of model systems that can be put to use examining the underlying mechanisms of pathogen transmission by ticks. Silicone membranes, in particular, offer researchers the flexibility to adjust membrane thickness and content during artificial feeding. The aim of the present study was to develop an artificial feeding technique using silicone-based membranes for all developmental stages of H. excavatum and H. marginatum ticks. Attachment rates after feeding on silicone membranes for females H. marginatum and H. excavatum were 8.33% (8/96) and 7.95% (7/88), respectively. The use of cow hair as a stimulant increased the attachment rate of H. marginatum adults in comparison to other stimulants. The engorgement of H. marginatum and H. excavatum females took 20.5 and 23 days with average weights of 307.85 and 260.64 mg, respectively. Although both tick species could complete egg-laying, and this was followed by hatching of larvae; their larvae and nymphs could not be fed artificially. Taken together, the results of the present study clearly indicate that silicone membranes are suitable for feeding of H. excavatum and H. marginatum adult ticks, supporting engorgement, laying of eggs, and hatching of the larvae. They thus represent a versatile tool for studying transmission mechanisms of tick-borne pathogens. Further studies are warranted to examine attachment and feeding behaviours in order to increase the success of artificial feeding of larvae and nymphal stages.

Evaluation of Cases with Suspected Canine Leishmaniasis History: A Five-year Retrospective Study (2016-2021)

Objective: This study aimed to evaluate the polymerase chain reaction (PCR) and immunofluorescence antibody test (IFAT) results of suspected samples with canine leishmaniasis (CanL) that were sent to the Parasitology Department Laboratories of the Veterinary Faculty in Aydin Adnan Menderes University. Methods: The age, gender, and breed of the dogs to be evaluated for CanL were recorded, and IFAT was performed using 80 blood serum samples collected from them. Additionally, after the isolation of genomic DNA of 27 blood samples, PCR of these samples was performed using primers that amplify the 145 bp kDNA region of Leishmania species. Results: Thirty-seven (46.25%) of the serum samples were seropositive in at least one dilution (1/64 or 1/128) according to IFAT. Five (18.5%) of the twenty-seven samples were positive for Leishmania DNA according to PCR. According to IFAT, 38.7% of male dogs and 59% of female dogs were positive. The highest number of seropositive samples were detected in dogs aged 3-5 years (11/27). Conclusion: Considering the zoonotic potential of leishmaniasis, which is considered endemic in the region, and the high positivity of the IFAT/PCR results, veterinarians should use advanced diagnostic methods, especially serological and molecular tests, in dogs with suspected CanL. The data obtained show that the risk of infection caused by Leishmania spp. is high in the region. Therefore, it is important to routinely ensure the control of CanL to protect both human and animal health.

Evaluation of Orally Administered Anthelmintic Treatment Options for Dentostomella translucida in Naturally Infected Mongolian Gerbils (Meriones unguiculatus)

Objective: This study aimed to evaluate the efficacy of eprinomectin, moxidectin and fenbendazole for treating Dentostomella translucida infections in naturally infected Mongolian gerbils (Meriones unguiculatus). Methods: A total of 28 gerbils were placed in individually numbered cages to determine the individual animal parasite load. Eggs per gram (EPG) counts were used to estimate the efficacy of the drugs. The day before the anthelmintic administration was denoted as day 0, and the EPG counts were determined by the McMaster technique from the stool removed from the cage bottom on days 7, 14, 21 and 28. The animals were assigned to one of four treatment groups according to their day 0 EPG counts. The orally administered drugs in the treatment groups were eprinomectin (15 mg/kg), moxidectin (0.4 mg/kg) and fenbendazole (12 mg/kg) for groups 1-3, respectively. The fourth group served as the control (without any drug administration). Results: Treatment efficacy was evaluated based on weekly EPG counts. The values decreased to zero in the fenbendazole group at 4 weeks of follow-up after treatment, and no parasite was found in any of the repeated examinations. The eprinomectin and moxidectin groups exhibited a fluctuating EPG state on both individual and group basis. Conclusion: D. translucida, which is known as the specific parasite of gerbils, can easily affect other members of the animal colony; thus, the control of its presence in gerbil breeding units is necessary. Therefore, the reported effective drug treatments are important for the fight against the investigated parasitic infection.

Investigation of Phlebotominae (Diptera: Psychodidae) Fauna, Seasonal Dynamics, and Natural Leishmania spp. Infection in Mugla, Southwest of Turkey.

Due to its geographical location, Mugla province is one of the most frequently used places by refugees. Although leishmaniasis have been previously reported in this region, there is a lack of information on the etiological agent and possible vectors. The main objectives of this study were; i) to investigate the sand fly fauna, ii) to reveal the natural Leishmania spp. infection in wild caught sand flies using molecular tools, and iii) to determine the annual seasonal dynamics of the sand flies in Mugla region. Totally, 2093 specimens belonging to 15 species [12 Phlebotomus, three Sergentomyia; 51 unidentified] were collected during the one-year (June 2016- June 2017) period. Of the collected sand flies, 1928 (92.12%) were caught by the Centers for Disease Control (CDC) light traps, while 165 (7.88%) of them were caught by sticky traps. Phlebotomus major sensu lato (s.l.), the potential vector of visceral leishmaniasis (VL) and canine leishmaniasis (CanL) in the Mediterranean and Aegean region, was detected in all sampling locations and found as the dominant taxon (n=1035; 49.45%) of the study area and followed by Phlebotomus tobbi (n=371; 17.72%). During the sampling period, sand fly activity was started in March and peaked in August. Sand fly population size reduced dramatically between mid-September and early October. The number of collected specimens was peaked in August, while there is only one sample collected both in November and March. The majority of the sand flies (78.66%) were collected at an altitude range of 200-400 m. Seventy-two monospecific pools were screened for the presence of Leishmania DNA by real time ITS1 PCR and 24 (nine P. major s.l., eight P. tobbi, two P. papatasi, two S. minuta, one P. alexandri, one P. similis, and one Phlebotomus (Transphlebotomus spp.) of them (33.8%) were found positive (L. infantum, L. tropica, and L. major). To the best of our knowledge, the presence of fifteen sand fly species and their distribution, seasonal dynamics, molecular detection of Leishmania parasites in Mugla province was reported for the first time. The presence of vector species in the study area, appropriate temperature and humidity conditions, long sand fly activity season, and presence of Leishmania parasite suggests that there is a serious risk in the transmission of leishmaniasis in Mugla.

Comparison of protectiveness of recombinant Babesia ovis apical membrane antigen 1 and B. ovis-infected cell line as vaccines against ovine babesiosis.

Babesiosis is a disease complex caused by unicellular Babesia parasites and among them, malignant ovine babesiosis caused by B. ovis has a devastating economical impact on the small ruminant industry. The control of disease is mainly based on chemotherapy and preventing animals from tick infestation and to date no vaccine is available against ovine babesiosis. The requirement for vaccination against B. ovis infection in endemically unstable regions is necessary for implementation of effective disease control measures. The aim of the present study was to evaluate the effectiveness of different immunisation protocols against disease in sheep experimentally vaccinated with recombinant B. ovis apical membrane antigen-1 (rBoAMA-1) and/or live, a B. ovis-infected cell line. Sheep were divided into four experimental groups, plus a control group. Animals were immunised either with the B. ovis stabilate, or with rBoAMA-1, or with both rBoAMA-1 and the B. ovis stabilate. Western blots and ELISAs indicated that immunisation with rBoAMA-1 resulted in generation of a specific response against the recombinant protein, but the degree of antibody response did not correlate with the level of induced protection against challenge. The strongest immune response was induced in animals co-immunised with the live B. ovis stabilate plus rBoAMA-1. Both the hematological and parasitological findings indicated that this co-immunisation regimen has vaccine potential to limit losses incurred by ovine babesiosis in endemic countries.

Vector and reservoir surveillance study in a canine and human leishmaniasis endemic area in most western part of Turkey, Karaburun.

Leishmaniasis is an arthropod borne disease that is endemic in 102 countries and one and half million new cases are reported each year. Sand flies are the one and only proven vectors of the disease and dogs are the main reservoirs in urban areas. Karaburun peninsula is located in most western part of Turkey and is reported to be an endemic area for human and canine leishmaniasis. The most recent study was undertaken more than 15 years ago in The peninsula and no clear data available for vectors or reservoirs. Thus, we aimed to update the information regarding sand fly diversity, infection status of reservoirs and vectors in the study area. Sand flies were collected using CDC light traps at 13 different sites of Karaburun and species identification was made using previously published keys. Monospecific pools were generated using midguts with blood retention and were screened for the presence of Leishmania spp. DNA by molecular techniques. A non-invasive conjunctival swab sampling was performed to identify the infection status among reservoirs and species typing of the causative agent was also undertaken using ITS1 PCR. Three out of 30 pools were found positive for Leishmania infantum that were generated using guts of Phlebotomus tobbi (n:36). Among all sampled dogs (44) and cats (19), 11 and one of them were found positive for L. infantum, respectively. There was a decrease in the number of P. papatasi during the study period, while increase was observed in the number of P. tobbi. The presence of proven vectors and reservoirs as well as Leishmania DNA in cats was shown in the present study. Sand fly fauna is updated and Leishmania DNA presence in cats was reported in the study area for the first time.

Dentostomella translucida (Gerbil Pinworm) Infection in Mongolian Gerbil (Meriones Unguiculatus) Schulz and Krepkorgorskaja, 1932.

The popularity of Mongolian gerbils (Meriones unguiculatus) as pets as well as experimental animals is continuously increasing. Mongolian gerbils are the main natural hosts of the nematode Dentostomella translucida, also referred to as pinworm, threadworm, or seatworm. D. translucida (Oxyuridae) was recently detected in the necropsy of a gerbil housed at the experimental animal production center of Adnan Menderes University Faculty of Veterinary Medicine. Mongolian gerbils are the main natural hosts of D. translucida, also referred to as pinworm. During necropsy in this animal, 26 parasites were collected from the small intestine and microscopically examined. The length of the female and male parasites was determined as 15.6-25.4 (mean, 18.3) and 10.2-16.8 (mean, 13.3) mm, respectively. The mean esophageal length in the female and male parasites was 397.3 and 325.3 mµ, respectively. The vulva of the females was close to the front end and was approximately 8.7 mm in length. The males had a single spiculum of approximately 342 mµ in length. There were seven papillae around the cloaca. During postmortem examination, eggs were also detected in feces collected from the gerbil's rectum. The eggs were spindle-shaped, slightly asymmetric, and had a diameter of 117-128 × 45-49 mµ (mean, 120 × 48 mµ). D. translucida, which is a parasite specific to gerbils, may easily affect other members of the animal colony. Thus, controling its presence in gerbil breeding units is essential.

Eustigmaeus johnstoni Zhang - Gerson (Acari: Stigmaeidae) a Parasitic Mite Species Detected in Phlebotomus papatasi (Diptera: Psychodidae) Specimens Collected from Aydin, Turkey.

OBJECTIVE: We aimed to report the presence of the parasitic mite Eustigmaeus johnstoni Zhang - Gerson (E. johnstoni) on the sand fly species Phlebotomus papatasi (P. papatasi) collected in Aydin province, Turkey. METHODS: Sand flies were collected from nature by CDC light traps in Nazilli town in Aydin province and were separated according to collection localities. Head and genital areas were cut and mounted for species identification, and midgut contents were checked for the possible presence of Leishmania parasites under a light microscope. Mites detected in sand flies were diagnosed on the species level based on the literature. RESULTS: A total of 360 female and 378 male sand flies were caught. During dissection and preparation, two female E. johnstoni parasitic mite specimens belonging to the genus Eustigmaeus were detected on the abdominal pleura of P. papatasi specimens. CONCLUSION: To our knowledge, this is the first study to report the presence E. johnstoni on P. papatasi in Turkey. Previous studies showing the relationship between parasitic mites and sand flies have indicated that this relationship is not accidental. More studies are needed to understand this relationship for obtaining more detailed information.