Sami Gökpinar

In-vivo efficacy of toltrazuril on experimentally induced Toxoplasma gondii tissue cysts in lambs: A novel strategy for prevention of human exposure to meat-borne toxoplasmosis

The aim of the present study was to investigate in vivo efficacy of toltrazuril on Toxoplasma gondii tissue cysts following induction of chronic toxoplasmosis in 4-week-old lambs (n=27) by inoculation of 1×10(5) T. gondii ME 49 strain oocysts (day 0). Beginning at the 15th day after inoculation, lambs in Group T20 and Group T40 were given toltrazuril orally 2 times, once every week (Baycox 5%, Bayer Animal Health) at a dose of 20 mg/kg and 40 mg/kg, respectively. Positive control (PC) lambs were not given any therapy, and 2 clinically healthy non-infected lambs were used as negative controls (Group NC). Two out of 9 lambs in PC group (oocyst inoculated but non-treated) were killed on toltrazuril treatment days (day 15 and 22) to evaluate the tissue cyst presence in their muscles. On day 90, the remaining 25 lambs were necropsied, and samples from the brain and 11 different muscle groups were collected. The tissues were examined for the presence of tissue cysts by histopathology, immunohistochemistry, nested-PCR and percoll gradient centrifugation. Anti-T. gondii antibodies were screened by IFAT throughout the experiment. The increased T. gondii seropositivity beginning from the 15th day of inoculation remained steady at Day 45 and Day 90 in Groups PC while it was significantly lower at Day 90 in toltrazuril receiving groups. In toltrazuril treated groups, histopathological findings included degenerative changes in the cyst wall, complete macrophage invasion to the cysts, and reduction or removal of the cysts in toto. Four out of 9 lambs (44.4%) in both toltrazuril treated group (Group T20 and T40) did not contain tissue cyst in any examined tissues while all positive control animals had T. gondii tissue cysts at least in one muscle group. The toltrazuril treatment efficacy on the cyst presence was determined as 44.4%. The number of the cysts in the musculature was significantly different between non-treated and toltrazuril treated lambs (X(2)=6.613; p=0.037). For the total number of cysts, the positive control lambs had higher number of cysts compared to both toltrazuril treated lambs (T20 and T40) (X(2)=5.629; p=0.018 and X(2)=5.629; p=0.018, respectively) while there were no differences between Group T20 and Group T40 (X(2)=0.000; p=1.000). According to PCR results, the brain and M. semitendinosus were positive in all 7 control lambs while 12 out of 18 lambs were positive in toltrazuril treated lambs. In conclusion, the results are promising as the toltrazuril treated lambs had markedly less parasite counts compared to those of untreated lambs. Further research should be conducted to reveal if toltrazuril treatment in sheep could be used as a strategy to minimize the cyst exposure of humans through consumption of raw or undercooked mutton.

Antiparasitic efficiency of Artemisia absinthium on Toxocara cati in naturally infected cats.

OBJECTIVE The first aim of the present study was to determine the efficiency of A. absinthium extract on cats naturally infected with Toxocara cati. The second aim was to determine the efficiency of the extract on the embryonic development of T. cati eggs in vitro. METHODS Artemisia absinthium extract was orally administrated to cats at the doses of 300 mg/kg and 600 mg/kg body weight in Group 1 and 2, respectively. It was given only once a day and the treatment continued 7 consecutive days. The faeces of the cats were examined both macroscopically and microscopically by flotation procedure with saturated salt solution pre-, during and post- treatment period. The faecal analysis was maintained during 8 days after completing the extract administration. The alteration of faecal egg numbers was performed by using the McMaster technique. RESULTS The faecal egg numbers per gram were decreased gradually in cats in the trial groups. In the treatment period, the activities of ALT, AST, ALP, urea and creatinine were located within the physiological ranges in cats. In in vitro trials with A. absinthium extract, the embryonic development of T. cati eggs was identical in all groups (treatment and control). A. absinthium extract did not inhibit larval development in eggs in in vitro trials. CONCLUSION This plant extract may be an alternative choice in the treatment of parasitic diseases in future.

Prevalence of Intestinal Parasites in Hamsters and Rabbits in Some Pet Shops of Turkey

OBJECTIVE In this study, we aimed to determine the parasite species carried by hamsters and rabbits purchased from some commercial pet shops in Turkey. METHODS For this purpose, the fecal samples of clinically healthy Syrian hamsters, dwarf hamsters, and crossbred rabbits were collected from 22 pet shops randomly selected in Ankara and Kirikkale provinces, located in Central Anatolia Region of Turkey. The fecal samples were examined with centrifuge flotation technique using saturated salt solution. RESULTS Parasitic infection rate was 57.5% in dwarf hamsters, 54.9% in Syrian hamsters, and 56.3% in crossbreed rabbits. Trichurid eggs were the most prevalent parasite in the feces of Syrians hamsters (28.1%). The other parasites of Syrian hamsters were as follows: Eimeria spp. oocysts (15.4%) and the eggs of H. nana (11.2%), Syphacia spp. (11%). and Aspiculuris spp. (5.6 %). Only trichurid eggs were observed in the fecal samples of dwarf hamsters (51.5%). Oocysts of Eimeria spp. (52.7%) and the eggs of P. ambiguus (3.6%) were detected in the feces of rabbits. CONCLUSION Within the scope of this study, the detection of H. nana eggs, a zoonotic parasite, in the feces of Syrian hamster was quite remarkable for public health.

Role of NETs in the difference in host susceptibility to Toxoplasma gondii between sheep and cattle

The main aim of this study was to compare extracellular traps (NETs) formation by polymorphonuclear neutrophils (PMNs) of cattle and sheep when exposed to T. gondii tachyzoites in vitro. The effects of parasite concentrations and different incubation periods on NETs development in cattle and sheep PMNs were studied. The effect of NET structures on host cell invasion by tachyzoites was also studied. This is the first report of NETs development by sheep and cattle PMNs against T. gondii in vitro. T. gondii-induced extracellular DNA production from PMNs was dependent on tachyzoite concentrations and incubation time in both sheep and cattle. Many nuclear and cytoplasmic changes were observed in sheep and cattle PMNs after exposure to T. gondii tachyzoites. The typical appearance of NETs, with MPO, NE and histone (H3) attached to extracellular DNA, was observed. Tachyzoites were entrapped within this structure. Myeloperoxidase (MPO) activity was higher in the cattle PMN-tachyzoite co-cultures than sheep. NETs structures released from sheep PMNs caused mechanical immobilisation of T. gondii tachyzoites, however, NET structures released from cattle PMNs may be lethal to tachyzoites. Bovine MPO may have a lethal effect on T. gondii tachyzoites in vitro during a 3h incubation. Besides other mechanisms that effect on host susceptibility to T. gondii in sheep and cattle, extracellular traps formation as a part of immunological reactions may be play a role in host susceptibility to T. gondii.

The relationship between seropositivity and tissue cysts in sheep naturally infected with Toxoplasma gondii

Skeletal muscles (tongue, masseter, leg, intercostal, and diaphragmatic muscles) and brain samples of 100 sheep at slaughter were analysed for the presence of T. gondii tissue cysts along with serum IgG titres. Two methods of isolation by percoll gradient centrifugation and tissue microarray (TMA) technique with immunoperoxidase staining were used. Seropositivity was detected in 88% (88/100) of sheep sera analysed by indirect fluorescent antibody test. Tissue cysts were observed in 46 (52.3%, 46/88) and 15 (17%, 15/88) of the seropositive sheep with the isolation technique and TMA and immunoperoxidase staining, respectively. The diameters of the tissue cysts were 25–58 × 25–62 (mean 34 × 36) µm. The relationship between the presence of tissue cysts and seropositivity in sheep was statistically significant at 1/16 (P < 0.01) and at 1/64 and 1/128 (P < 0.001) serum dilutions.

Prevalence of Intestinal Parasites in Cats and Their Importance in Terms of Public Health

OBJECTIVE This study aimed to detect the prevalence of intestinal parasites in cats by fecal examination. METHODS A total of 100 fecal samples were collected from owned and stray cats in the Kırıkkale province. Epidemiological data on the sampled cats (e.g., age, race, and sex) were recorded. The samples were macroscopically investigated for the presence of cestode proglottids. The fecal smears were stained with Giemsa and Carbol fuchsin stains. The samples were prepared by centrifugal flotation with a saturated sugar solution. The slides were examined using light microscopy. RESULTS Parasites were detected in 47% of feces of cats examined. Protozoa (Isospora spp. and Cryptosporidium spp.) oocysts, gravid proglottid of cestode (Joyeuxiella spp.), and nematode (Toxocara spp. and hookworm) eggs were present in the fecal samples. In this study, 48.9% of cats were infected with one species, and 44.6% and 6.3% of cats were infected with two and three species, respectively. The more prevalent parasite species were Isospora spp. (65.9%) and Toxocara spp. (48.9%). The parasites were found to be more common in stray cats. CONCLUSION Intestinal parasites were highly prevalent in the cats examined in this study. House cats can have significant parasitic infections, particularly because they are allowed outdoors. House cats should be regularly examined by a veterinarian.

Seroprevalence of Neospora caninum in Dairy Cattle Raised in Çiçekdaği District of Kırşehir Province

OBJECTIVE Neospora caninum is one of the most important causes of abortion in cattle worldwide and causes significant economic losses in the meat and dairy industries. This study aimed to determine the seroprevalence of N. caninum in dairy cattle raised in Çiçekdağı district of Kırşehir province. METHODS One hundred sixteen serum samples collected from dairy cattle were analyzed for N. caninum antibodies by a commercial Enzyme-Linked ImmunoSorbent Assay (ELISA) kit (VMRD c-ELISA). RESULTS The seropositivity rate was 18.1% in the cattle examined. The seroprevalence of N. caninum was 23.4% in dairy cattle with fertility problems, 33.3% in cattle with a history of abortion, and 7.8% in clinically healthy dairy cattle (p=0.006). Cattle breeds with highest seropositivity rates were Holstein, Simmental, and Brown Swiss (p=0.008). CONCLUSION Control measures should be taken for both dogs as final host of the parasite cattle to prevent the spread of neosporosis in cattle in Çiçekdağı district.

Prevalence of Haemoproteus spp. in Tumbler Pigeons (Columba livia domestica) in Kirikkale Province, Turkey

OBJECTIVE Haemoproteus spp. are common blood parasites of pigeons. They have been reported in pigeons in many regions worldwide, including Turkey. Pigeon breeding is a popular hobby in Kirikkale province, and there is no information about the prevalence of Haemoproteus spp. The present study aimed to determine the prevalence of Haemoproteus spp. in tumbler pigeons in Kirikkale province (Kırıkkale and Yahsihan district). METHODS Blood samples were taken from the wing vein of pigeons (n: 173) through microcapillary (with/heparin) tubes between February and March 2016. Blood smears were stained with 5% Giemsa solution. Ectoparasites of the pigeons were collected in separate sealed boxes. Epidemiological data of the sampled pigeons (age and sex) were obtained from the breeders. RESULTS In total, 23 (%13.2) of 173 pigeons were infected with Haemoproteus spp. Parasite was detected in 73.9% of pigeons over 1 year old and 26.1% of pigeon under 1 year age. Haemoproteus spp. was observed in 56.2% of females (13/23) and 43.4% of males (10/23), Sex-related differences were not observed (p = 0.821). Ectoparasites of the pigeons were identified as Columbicola spp. CONCLUSION To the best of our knowledge, this is the first study in Kirikkale province that reported the prevalence of Haemoproteus spp. in pigeons.