Molekularna detekcija ehrlichia canis u populaciji pasa -kućnih ljubimaca u Severnoj Makedoniji

Canine monocytic ehrlichiosis (CME) is a widespread, tick-borne, canine disease, caused by an obligate intracellular bacterium, Ehrlichia canis. The main vector, a brown-dog tick, Rhipicephalus sanguineus, is widely distributed, especially in areas with tropic, subtropic, or Mediterranean climates (Central and South America, Eastern and Western Asia, Africa, Australia and Southern Europe). The study performed in 2012, by Stefanovska et al., determined a seroprevalence of 18.7% of E. canis among the Macedonian dog population. Up to date, the presence of E. canis, using molecular diagnostic methods, has not been investigated in Macedonia. Therefore, this study aimed to confirm the presence of E. canis, in the pet-dog population on the territory of the city of Skopje, North Macedonia, using a highly sensitive multiplex Real-Time PCR method (qPCR). Whole blood samples from 80 dogs of different breeds and ages, with clinical symptoms of CME and positive serology result for the presence of antibodies against E.canis, were collected for analyses. Out of 80 dogs, 36 (45%) were found as positive. The present work reports the first molecular detection of E. canis in pet dogs on the territory of the city of Skopje, Macedonia.Monocitna erlihioza pasa (CME) je široko rasprostranjena bolest pasa, koja se prenosi krpeljima, a uzrokuje je obligatno-intracelularna bakterija Ehrlichia canis. Glavni vektor, smeđi krpelj pasa, Rhipicephalus sanguineus, široko je rasprostranjen, posebno u oblastima sa tropskom, subtropskom ili mediteranskom klimom (Centralna i Južna Amerika, Istočna i Zapadna Azija, Afrika, Australija i Južna Evropa). U Studiji izvedenoj 2012. godine, Stefanovska i saradnici su utvrdili da među severnomakedonskom populacijom pasa, seroprevalencija E. canis iznosi 18,7%. Do danas, prisustvo E. canis, koristeć i molekularne dijagnostičke metode, nije istraženo u Severnoj Makedoniji. Stoga je ova studija imala za cilj da potvrdi prisustvo E. canis u populaciji kuć nih ljubimaca na teritoriji grada Skoplja u Severnoj Makedoniji, koristeć i visoko osetljivu multiplex Real-Time PCR (qPCR). Za analize su prikupljeni uzorci pune krvi od 80 pasa različitih rasa i uzrasta, sa kliničkim simptomima CME i pozitivnim serološkim rezultatom na prisustvo antitela protiv E.canis. Od 80 pasa, 36 (45%) je ocenjeno kao pozitivno. Ovaj rad izveštava o prvom molekularnom otkrivanju E. canis kod pasa kuć nih ljubimaca na teritoriji grada Skoplja, Severna Makedonija

Serological profile and pleurisy lesions associated with Actinobacillus pleuropneumoniae in pig farms in North Macedonia

Actinobacillus pleuropneumoniae (App) is one of the most important swine respiratory pathogens that causes porcine pleuropneumonia and massive financial losses in pig industry. The objective of this study was to investigate App infection in five pig herds from North Macedonia experiencing clinical respiratory infections by serological testing and a slaughterhouse pleurisy evaluation system (SPES). In total, 250 blood samples were taken from pigs aged 6, 10, 14, 18, and 22 weeks. Ten animals per age category from each farm were sampled and analyzed for presence of antibodies against App. At the slaughterhouse, 50 lungs per herd from slaughtered age pigs were scored by the SPES for the presence of lesions associated with chronic pleurisy. The overall seroprevalence to App was 65.6%, ranging from 34% to 98% at the farm level. The highest seroprevalence was found in 6-week-old pigs in all farms, while significant differences were observed between farms in the 10-, 14-, 18-, and 22-week-old pig groups. Pleurisy associated with App was found in 26.4% of all examined lungs, with the mean SPES score being 0.75 (0.14 - 1.10). The percentage of SPES scores of 0, 1, 2, 3, and 4 in all lungs were 73.4%, 1.6%, 8.8%, 8.4%, and 7.6% respectively. A significant difference in mean SPES score was obtained between two farms. High seroprevalences of App detected on the tested farms were probably due to constant natural infection. The highest seroprevalences, measured in the youngest pigs, could be due to maternallyderived antibodies. Higher seroprevalence against App and lower SPES scores in some farms suggests immunity resulting from infection by corresponding field serotypes

Immunohistochemical investigation of FIPV3-70 antigen expression in the ileum of cats with effusive feline infective peritonitis

One of the most common infectious causes of cat mortality is feline infective peritonitis (FIP), along with panleukopenia and viral upper respiratory tract infections. FIP is more likely to affect cats whose immune system is weak or suppressed. It is thought that the infection of macrophages and monocytes plays a major role in the pathogenic process. In order to set a definitive diagnosis for this infectious disease, a histopathological examination of tissues, and feline coronavirus (FCoV) detection by immunohistochemistry (IHC) is necessary. In this investigation, 15 cats between the ages of 5 and 24 months with clinical suspicion of FIP, underwent post-mortem necropsy, pathohistological and immunohistochemical examination. The results showed that all the cats had abdominal effusion with pyogranulomas throughout the abdominal serosa. Ten out of fifteen cats were FIP positive using immunohistochemical methods. This method also showed the antigen deposition in the macrophages thus confirming their role in the pathogenesis of FIP

First case of Strongyloides stercoralis infection in a dog in the Republic of Macedonia

A six-month-old Pomeranian male dog was referred due to a month long history of unformed, soft faeces and mild weight loss. Stool analyses by direct faecal smear, Zinc sulphate flotation and the Baermann concentration method revealed an infection with Strongyloides stercoralis. The dog was initially treated once with a combination drug of praziquantel, pyrantel and febantel (½ Drontal® Plus Tablets for puppies and small dogs; Bayer; i.e. 31.5 mg/kg bodyweight of febantel). The treatment was repeated after 12 days with the same dosage for 3 consecutive days. The stool analyses performed 14 days and 3 months after the second treatment were negative for S. stercoralis larvae. The results suggest that a repeated treatment with Drontal® Plus Tablets is effective against S. stercoralis in dogs and has no adverse effects

Immunohistochemical detection of estrogen receptors in canine mammary tumors

Mammary tumors are among the most common neoplasms in intact female dogs.They have a complex morphology, usually affecting middle age and older bitches. Almost 50% of the mammary tumors in dogs are malignant neoplasms. Prognosis is based on several factors: stage, age, tumor size, metastasis, histopathology, ovariectomy status and hormone-receptor activity. Immunohistochemical (IHC) measurement has become increasingly an important diagnostic and prognostic parameter, with the development of monoclonal antibodies against nuclear estrogen and progestin receptors. The aim of this study was to detect the presence of ER receptors in malignant canine mammary tumors and to identify their association with the clinical course of the tumor. Mammary tumor samples have been obtained by mastectomy from dogs presented at our clinic. Detailed clinical examination, CBC and basic serum biochemical profile were performed in all patients. Surgery was the only treatment. Histopathological examination and immunohistochemical detection of estrogen α receptors (ERα) was performed on 8 formalin-fixed, paraffin-embedded tissue samples, using the PT LINK immunoperoxidase technique. Histopathological examination of the mammary tumor samples (n=11) revealed tubular adenocarcinoma (n=6,54.5%) and ductal adenocarcinoma (n=3, 27.3%), one patient with benign adenoma and one with mastitis. Patients with positive ER tumors are alive, without remission, while 3 of the patients that were ER negative died due to lung metastases. According to our results, it can be concluded that the appearance and development of canine mammary tumors is highly connected with ovarian steroid hormones and that immunostaining of the tumors may be used as a good prognostic parameter in these patients

Effects of doxycycline treatment on hematological and blood biochemical parameters in dogs naturally infected with Ehrlichia canis

Ehrlichia canis, the etiologic agent of canine monocytic ehrlichiosis (CME), is mainly transmitted by the brown dog tick Rhipicephalus sanguineus. Clinical signs of the disease can be various, depending on the stage of the disease. Typical changes in hematological and blood biochemical parameters are: severe thrombocytopenia, mild to marked non regenerative anaemia and hypoalbuminemia. In order to present the effects of the treatment protocol on several hematological and biochemistry parameters, 34 Ehrlichia canis positive dogs were compared before and after treatment with doxycycline 10mg/kg/day, in duration of four weeks. Besides the clinical sings and laboratory findings, diagnosis was confirmed by antibody tests (Bionote, Korea, AGROLABO S.p.A., Italy). The most common clinical sings were depression, lethargy, pyrexia, vomiting and anorexia. Hematological analyses were performed on the automatic hematology analyzer Exigo ЕоѕVet (Sweden), while biochemistry analyses (alanine aminotransferase, aspartate aminotransferase, urea, creatinine, albumin, total protein, globulin and alkaline phosphatase) were performed using the automatic analyzer ChemWell 2910 (Awareness Technology, INC, USA). Statistically significant difference (p<0.05) in hematology changes was present regarding the red blood cells count, platelet count, hematocrit and hemoglobin before and after treatment. Hypoalbuminaemia (Mean 19.21 ±4.96 g/l) was the only serum biochemistry parameter with significant change before and after treatment, as well. Treatment with doxycycline in patients with E.canis resulted in significant increase of hematology parameters (red blood cells, hemoglobin, haematocrit and platelets), as well as albumins in serum

IMMUNOHISTOCHEMICAL DETECTION OF ESTROGEN RECEPTORS IN CANINE MAMMARY TUMORS

ABSTRACT Short Communication Mammary tumors are among the most common neoplasms in intact female dogs.They have a complex morphology, usually affecting middle age and older bitches. Almost 50% of the mammary tumors in dogs are malignant neoplasms. Prognosis is based on several factors: stage, age, tumor size, metastasis, histopathology, ovariectomy status and hormone-receptor activity. Immunohistochemical (IHC) measurement has become increasingly an important diagnostic and prognostic parameter, with the development of monoclonal antibodies against nuclear estrogen and progestin receptors. The aim of this study was to detect the presence of ER receptors in malignant canine mammary tumors and to identify their association with the clinical course of the tumor. Mammary tumor samples have been obtained by mastectomy from dogs presented at our clinic. Detailed clinical examination, CBC and basic serum biochemical profile were performed in all patients. Surgery was the only treatment. Histopathological examination and immunohistochemical detection of estrogen α receptors (ERα) was performed on 8 formalin-fixed, paraffin-embedded tissue samples, using the PT LINK immunoperoxidase technique. Histopathological examination of the mammary tumor samples (n=11) revealed tubular adenocarcinoma (n=6,54.5%) and ductal adenocarcinoma (n=3, 27.3%), one patient with benign adenoma and one with mastitis. Patients with positive ER tumors are alive, without remission, while 3 of the patients that were ER negative died due to lung metastases. According to our results, it can be concluded that the appearance and development of canine mammary tumors is highly connected with ovarian steroid hormones and that immunostaining of the tumors may be used as a good prognostic parameter in these patients