Arginine, symmetric and asymmetric dimethylarginine levels in canine leishmaniasis.

The study aimed to investigate the levels of arginine, symmetric dimethylarginine (SDMA), and asymmetric dimethylarginine (ADMA) in dogs with canine leishmaniasis (CanL) and their relationship with some renal and cardiovascular parameters. A total of 60 dogs were enrolled, including 40 with CanL and 20 healthy controls. The CanL group was divided into four stages based on clinical and laboratory findings. The levels of plasma arginine, SDMA, and ADMA were determined by high performance liquid chromatography (HPLC). The data from the healthy group were compared with those from the CanL group, and according to the stages. In dogs with CanL, systolic and diastolic blood pressure, plasma creatinine, cystatin-C, phosphorus, potassium, and low-density lipoprotein concentrations, the urine protein/creatinine ratio, the amount of nitric oxide, and creatine kinase-MB activity were higher, while the high-density lipoprotein concentration was lower compared to healthy controls. The concentration of arginine was low (p < 0.05) and the levels of ADMA (p < 0.001) and SDMA (p < 0.05) were high in dogs with CanL. There were no statistically significant differences in arginine concentration among the different stages of CanL. However, the concentration of plasma ADMA was higher in all stages of CanL compared to the healthy group, and the concentration of plasma SDMA was higher in Stage IV compared to the healthy group and Stage III. The present study demonstrates for the first time a decrease in arginine concentration and an increase in ADMA concentration in dogs with CanL. The increase in SDMA concentration in dogs with CanL was consistent with previous studies. However, compared to other renal parameters, SDMA exhibited limited performance distinguishing between clinical stages of CanL. These findings could be a source for future diagnostic and therapeutic studies to explain the renal and cardiovascular pathophysiology of CanL. Additional clinical studies that include treatment and patient follow-up with an assessment of the acute phase response are needed to provide a more detailed understanding of the changes observed in dogs with CanL.

Oxidative Status and Lipid Profile among Dogs at Different Stages of Visceral Leishmaniasis.

OBJECTIVE: The purpose of the study was to investigate the oxidative status and lipid profile among dogs at different stages of visceral leishmaniasis (VL). METHODS: Dogs with VL were divided into four groups according to the classification reported by the Leishvet group: stage I (n=9), stage II (n=11), stage III (n=6), and stage IV (n=6); these dogs were compared to healthy control dogs (n=14). The lipid profile [high-density lipoprotein (HDL), low-density lipoprotein (LDL), cholesterol, and triglyceride levels) and oxidative status [total antioxidant capacity (TAC), total oxidant status (TOS), and paraoxonase-1 (PON-1) activity] were evaluated. RESULTS: Compared to the control dogs, significant increases in the TOS and the LDL level and decreases in PON-1 activity and the HDL level were determined among the dogs at all stages of VL (p<0.05). No significant differences were found in the TAC and the cholesterol and triglyceride levels among the groups. CONCLUSION: Increased oxidative stress and alterations in lipid profile were observed among dogs with VL. However, no significant differences were detected between dogs at different stages of the disease. Therefore, changes in the TOS, PON-1 activity, and HDL and LDL levels in dogs at all stages of VL should be considered in the diagnosis of the disease and planning of the treatment.

Seroprevalence of Canine Leishmaniasis in Northern Cyprus.

OBJECTIVE: Canine leishmaniasis (CanL) is an important public and veterinary health problem in Mediterranean Basin countries. In this study, we aimed to determine the seroprevalence of CanL in several provinces of Northern Cyprus. METHODS: The seroprevalence of CanL was determined by the indirect fluorescent antibody (IFA) test in dog sera. In total, 281 dogs were randomly selected from Nicosia (n=80), Trikomo (n=58), Famagusta (n=60), Morphou (n=30), and Kyrenia (n=53), consistent with a statistically representative number of the regional dog population. RESULTS: Ten (3.55%) out of 281 dogs were found to be seropositive by the IFA test. CanL seropositivity differed between cities as follows: 1.72% (1/58) in Trikomo, 13.20% (7/53) in Kyrenia, 1.67% (1/60) in Famagusta, and 3.33% (1/30) in Morphou. No seropositive dog was found in Nicosia. The symptoms in 37 out of 281 dogs were generalized lymphadenopathy, weight loss, alopecia, exfoliative dermatitis, and epistaxis. Four out of 10 seropositive dogs showed at least one clinical symptom that could be related with CanL. CONCLUSION: CanL seroprevalence was found to be 3.55% (10/281) in Northern Cyprus. Seropositive dogs, in particular, had lived in areas that exhibited rural as well as urban characteristics.

Evaluation of conjunctival swab sampling in the diagnosis of canine leishmaniasis: A two-year follow-up study in Çukurova Plain, Turkey.

The diagnosis of canine leishmaniasis (CanL) in symptomatic and asymptomatic dogs is a very important and problematic public health issue in Turkey. A longitudinal study was carried out on dogs in selected villages in the Çukurova Plain in Turkey, from July 2011 to June 2013, where cutaneous (CL) and visceral (VL) leishmaniasis is endemic. The study aimed to determine the prevalence of CanL and to evaluate the early diagnostic performance of the non-invasive conjunctival swab nested PCR (CS n-PCR) test in comparison with the Indirect Fluorescent Antibody Test (IFAT). The consecutive blood and CS samples from a representative number of dogs (80-100 dogs/each survey) were collected in a cohort of 6 villages located in the area. Clinical symptoms, demographic and physical features about each dog were noted and lymph node aspiration samples were obtained from selected dogs with lymphadenopathy. In four surveys during the period, a total of 338 sets (blood and CS) of samples from 206 dogs were obtained, such that 83 dogs were sampled more than once. In the cross-sectional analysis, the CanL prevalence was found to be 27.18% (between 7.14% and 39.13%) by IFAT and 41.74% (between 29.03% and 46.66%) by CS n-PCR. The isolated strains were identified as Leishmania infantum MON-1 (n=9) and MON-98 (n=2) by MLEE analysis. Genetic studies targeting the Hsp70 and ITS1 regions performed on 11 dog isolates also showed two clear separate groups. According to IFAT results, 24 of the 83 dogs sampled more than once showed seroconversion (n=19) or a four-fold increase in Ab titers (n=5), while 17 were positive in the initial screening. Forty-two dogs stayed negative during the whole period. The natural Leishmania exposure rate was detected as 31.14% in the study area. CS n-PCR only detected Leishmania infection earlier than IFAT in 8 dogs. No statistical difference was found after the analysis of demographical and physical data. The results indicated that (i) circulation of the dog population is very common in settlements in the Çukurova Plain, but the disease prevalence is high and stable, (ii) the performance of CS n-PCR for detecting Leishmania-dog contact is higher than IFAT, (iii) and some of the parasites isolated from dogs have different zymodemes and/or genotypes from previous human and sand fly isolates; suggesting the probability of two different cycles of leishmaniasis in this particular area. This hypothesis should be supported by future studies targeting vectors and reservoirs.

Detection of Leishmania major and Leishmania tropica in domestic cats in the Ege Region of Turkey.

Leishmaniosis is a group of diseases caused by different species of Leishmania parasites in mammalian species. The aim of the present study was to investigate the presence of Leishmania spp. DNA in cats using real time polymerase chain reaction (RT-PCR) assays targeting internal transcribed spacer (ITS1) and heat-shock protein 70 gene (Hsp70) regions with Leishmania species-specific primers and probes. Blood samples were collected from 147 cats (73 female; 74 male) in the endemic regions for zoonotic visceral leishmaniasis in the western provinces of Turkey and analyzed using two RT-PCR assays. Additionally, Hsp70 RT-PCR products were sequenced. ELISA assays for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV) were also carried out for 145 of the 147 samples. Overall, 13/147 (8.84%) cats were positive for Leishmania by RT-PCR (4 L. major and 9 L. tropica). FIV and FeLV antibody and/or antigen was detected in 4 and 5 cats among Leishmania DNA positives, respectively. To the best of our knowledge, this study is the first to investigate and report the presence of L. major and L. tropica infections in a large group of domestic cats in Turkey. The results obtained indicate that species identification of Leishmania is essential for epidemiological understanding and that clinical signs alone are not indicative for leishmaniosis in cats, as it is in dogs. This study suggests that extensive research should be carried out in cat populations in order to fully understand the role of cats in the epidemiology of the disease.

A serological investigation of Bartonella henselae infection in cats in Turkey.

Bartonella henselae is the causative agent of cat scratch disease (CSD) in humans. Cats are the main reservoir of this bacterium and may infect humans through scratches and bites. The purpose of this study was to determine the B. henselae seroprevalence in cats in Turkey. A total of 298 cats blood samples were collected from six different provinces of Turkey. Sera were tested for the presence of anti-B. henselae IgG antibodies by indirect fluorescent antibody test (IFA). The seroprevalence of B. henselae was 27.9% (83/298) for the cats examined in this study. The seroprevalence of cats by province was significantly higher in Bursa (41.3%), Adana (33.9%), Aydin (27.5%) and Burdur (32.3%) than in Kayseri (17.9%) and Istanbul (12.5%). Statistically significant differences were not observed between cat sexes and living conditions of cats. The results revealed that B. henselae is an important zoonotic pathogen in Turkey.

Failure of combination therapy with imidocarb dipropionate and toltrazuril to clear Hepatozoon canis infection in dogs.

Current treatments with imidocarb dipropionate for infected dogs with Hepatozoon canis do not always provide parasitological cure. The objective of this study is to determine whether concomitant use of toltrazuril may potentiate the effect of imidocarb dipropionate in the management of H. canis infection (HCI). Twelve dogs were determined to have naturally HCI based on clinical signs, identification of the parasite in blood smears, and serologic assay. The animals were allocated randomly to one of two groups (n = 6 in each group). Dogs in Imi group were given imidocarb dipropionate at a dose of 6 mg/kg body weight subcutaneously in two injections 14 days apart. Imi plus Toltra group was given imidocarb dipropionate as dose mentioned above and toltrazuril at 10 mg/kg/day orally for the first five treatment days. Clinical findings, blood counts and parasitaemia levels in blood before and 14, 28 and 56 days after the initial treatment were performed to evaluate treatment response. The overall clinical efficacy of imidocarb dipropionate with and without toltrazuril was 83.3% and 66.7%, respectively; with a mean recovery time of 21.0 and 25.6 days, respectively. A substantial main effect of time on mean PCV, Hb, WBC, neutrophil and PLT and gradual reduction of parasitaemia were significantly observed in both groups (P < 0.05), whereas no significant difference was noticed between the studied protocols. The parasitologic cure rate at the end of eight weekly observation period was 16.6% and 33.3% in Imi and Imi plus Toltra groups, respectively. Similar clinicopathologic and parasitologic responses were observed in both treated groups; thus, it was concluded that toltrazuril does not reveal additional benefit to imidocarb therapy in dogs with HCI.

Seroprevalence of Bartonella vinsonii subsp. berkhoffii in urban and rural dogs in Turkey.

The seroprevalence of Bartonella vinsonii subsp. berkhoffii was investigated in stray urban dogs and shepherd and farm guard dogs from rural areas sampled from 10 provinces of Turkey. Sera from 855 dogs were examined for the presence of anti-B. vinsonii subsp. berkhoffii antibodies by indirect fluorescent antibody test. Overall, 56 (6.6%) of the 855 dogs examined, including 16 (3%) of the 522 stray dogs and 40 (12%) of the 333 rural dogs, were seropositive. This is the first report on prevalence of antibodies to B. vinsonii subsp. berkhoffii in dogs in Turkey.

[Investigating the seroprevalance of leishmaniasis in four dog shelters in Antalya and its districts]. / Antalya Ili ve Ilçelerindeki Dört Köpek Barinaginda Leishmaniasis Seroprevalansinin Arastirilmasi.

Human visceral leishmaniasis (HVL) is endemic along the Aegean and Mediterranean coasts, while it occurs sporadically in other regions of Turkey. A relationship between canine leishmaniasis (CanL) and HVL has been detected and dogs have been found to be the reservoir of Leishmania infantum. In this epidemiologic study, the seroprevalence of CanL was investigated in dog shelters of four districts (Kepez, Kemer, Alanya and Gazipasa) of Antalya, in which visceral leishmaniasis and cutaneous leishmaniasis were observed according to the official records of The Ministry of Health of Turkey. Blood specimens of 176 dogs were examined and seropositive, borderline, seropositive, and negative results were detected in 14 (7.95%), 24 (13.63%), and 138 (78.4%) dogs, respectively. Weight and alopecia, onychogryphosis, and skin ulcers around the nose were observed in only two (14.2 %) dogs of seropositive cases. In conclusion, our data indicate that there is a potential danger for humans because L. infantum carrying dogs were detected in four districts of Antalya in the present study and also the appropriate vector spp. for this parasite has been reported in a previous one. Therefore, it would be advisable to perform new studies in order to determine the seroprevalence of CanL in dogs and the population size of vector spp. of L. infantum in other districts of Antalya.

[Therapeutic efficacy of buparvaquone (buparvon) in cattle with theileriosis]. / Theileriosisli Sigirlarda Buparvaquone (Buparvon)'un Terapotik Etkinligi.

The aim of this study was to examine the efficacy of buparvaquone (Buparvon, ALKE, Istanbul) in the treatment of theileriosis in cattle. The causative agent T. annulata causes direct and indirect gross economical loss in Turkey. Theileriosis was microscopically diagnosed by determining the presence of piroplasms in erythrocytes in thin blood smears stained with Giemsa stain. Buparvaquone was administered intramuscularly to the cattle with theileriosis using a single dose of 2.5 mg/kg. Clinical and laboratory examinations in cattle with theileriosis were carried out before and on the first, third and seventh days after treatment. As a result, it was concluded that a single deep intramuscular dose of 2.5 mg/kg buparvaquone was effective in the treatment of cattle which are naturally infected with T. annulata. However, further studies are needed using control groups of the same breed and age including suitable numbers of naturally infected and experimentally infected cattle.

A parasitological, molecular and serological survey of Hepatozoon canis infection in dogs around the Aegean coast of Turkey.

Canine hepatozoonosis is caused by the tick-borne protozoon Hepatozoon spp. The prevalence of the infection in the Aegean coast of Turkey was investigated by examination of blood smear parasitology and polymerase chain reaction (PCR) using blood samples from 349 dogs collected from Central Aydin, Kusadasi, Selcuk, Central Manisa, Bodrum and Marmaris within the Aegean coast of Turkey. The indirect fluorescent antibody test (IFAT) for the detection of Hepatozoon canis antibodies was also used to detect the exposure rate to H. canis. PCR amplifying a 666bp fragment of 18S rRNA gene of Hepatozoon spp. was used in the epidemiological survey. The prevalence of Hepatozoon spp. infection was 10.6% by blood smear parasitology and 25.8% by PCR. IFAT revealed that 36.8% of serum samples were positive for antibodies reactive with Hepatozoon spp. The PCR products of 18S rRNA gene of Hepatozoon spp. isolated from six infected dogs, one isolate originating from each of the six different locations, were sequenced. The results of sequence analysis indicate that they are closely related to Indian and Japanese isolates of H. canis. This is the first epidemiological study on the prevalence of H. canis infection in the dog, in Turkey.

Dogs with Hepatozoon canis respond to the oxidative stress by increased production of glutathione and nitric oxide.

Canine hepatozoonosis is a disease caused by the tick-borne protozoan Hepatozoon spp. It has been reported in the United States, southern Europe, the Middle East, Africa and the Far East. In Turkey, canine hepatozoonosis was reported for the first time in 1933. In the present study, serum glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO) and ceruloplasmin levels were analysed in 14 dogs infected with Hepatozoon canis as well as in 10 healthy dogs. Blood smears were prepared from peripheral blood and ticks were collected for identification in the laboratory. Rhipicephalus sanguineus was found only on diseased dogs. No ticks were observed on healthy dogs. The diagnosis of H. canis is made mainly by the detection of gametocytes within neutrophils and monocytes. The haematological diagnosis was confirmed using PCR analyses by amplifying a partial 18S rRNA gene sequence of Hepatozoon spp. Infection was detected in 14 animals. Compared to controls, the serum GSH, MDA and NO levels in infected animals increased significantly (p<0.05, <0.01 for MDA), whereas the concentrations of ceruloplasmin in diseased animals remained unaltered. The results of the present study suggest that in dogs infected with H. canis increased levels of GSH, MDA and NO may be related to host's defences against parasitic infection.

Clinical and serological follow-up in dogs with visceral leishmaniosis treated with allopurinol and sodium stibogluconate.

Seven dogs with parasitologically proven clinical visceral leishmaniosis (Leishmania infantum infection) were treated with a combination of allopurinol and sodium stibogluconate. The dogs received first orally 15 mg/kg of allopurinol every 12 h until the clinical signs improved, in the following 1 month period allopurinol at same dose and subcutaneously 30 mg/kg of sodium stibogluconate combination were given daily and at the end of the combined treatment, allopurinol was continued alone at the same dose till the end of 8 months. During the treatment period, dogs were supported by additional proteins, vitamins, and minerals. A long acting insecticide (collar or drop) was also used in order to prevent further parasite transmission. Follow-up was maintained by clinical, clinicopathological evaluation, and parasitological examination of lymph node, serology using the indirect immunofluorescent antibody test (IFAT). Before treatment commenced, the most important clinical signs were exfoliative dermatitis, ulcerations, peripheral lymhadenopathy, pale mucous membranes, weight loss, and ocular lesions. Clinicopathological findings included commonly anaemia, hyperproteinaemia, hyperglobulinaemia and hypoalbuminaemia. Before the treatment, amastigotes were seen in six of the seven dogs by examination of lymph node aspiration, and IFAT-titers were positive in all dogs. At the end of 8 months treatment, remission of clinical signs, restoration to normal of clinicopathological abnormalities were noticed. Lymph node aspiration was performed on three out of the seven dogs at the end of the treatment because of the very small sizes of the lymph nodes, and no amastigotes were observed. Although the mean IFAT-titer of the dogs were significantly (P < 0.001) lower compared with pretreatment, IFAT-titers of dogs were still positive. No relapses occurred during treatment period and a 6-24-month duration after the end of therapy. Based on the above results, long-term use of allopurinol combined with sodium stibogluconate together with support treatment concluded to have enough therapeutic efficacies in the treatment of dogs with visceral leishmaniosis. Observations of the cases for possible relapses were still going on and insecticide application was carefully carrying on in order preventing a possible re-infection.

Serum and hair levels of zinc and other elements in dogs with visceral leishmaniasis.

The aim of this study was to determine the zinc, iron, copper, calcium, phosphorus, and magnesium levels in blood serum and zinc and copper levels in hair of dogs with canine visceral leishmaniasis. The serum zinc and iron levels were found to be significantly lower in diseased dogs than those of healthy controls. Serum copper levels were significantly higher, whereas no significant differences were observed for calcium, phosphorus, and magnesium. There were no significant differences in the zinc and copper levels in hair. Our results show that the serum zinc, iron, and copper levels are altered in canine leishmaniasis.