Kongenitalna kifoza u torakalnom delu kičme usled prisustva hemivertebre kod šteneta mopsa

Congenital spine deformation caused by vertebral malformation is reported in different brachycephalic dog breeds, but also in other large dog breeds. The most severe clinical signs are due to spinal cord compression. Depending on localisation of the lesion, clinical signs can manifest differently. A five-month-old pug was brought for autopsy to the Department of Forensic Veterinary Medicine, Faculty of Veterinary Medicine, University of Belgrade. The first clinical signs, including general weakness and difficulty to jump appeared when he was three and a half months old. They became more severe until the dog was four months old. Radiography and myelography showed severe kyphosis and spinal cord compression in the T8-T9 region. After euthanasia, an autopsy was performed. Pathomorphological examination showed severe deformation of the spine, kyphosis and a wedge-shaped T8 vertebra. Although, literature data indicates that clinical signs can be latent until five or six months of age, this case shows that even in young puppies, three to four months old, serious clinical signs can be present. This is of great clinical importance, not just for veterinarians, but also for brachycephalic dog owners and breeders.Kongenitalne deformacije kičme uzrokovane malformacijama na vertebrama javljaju se kod različitih brahicefaličnih rasa, ali se mogu javiti i kod velikih rasa pasa. Najteži klinički simptomi se javljaju usled kompresije kičmene moždine. U zavisnosti od lokalizacije lezije, simptomi se mogu različito manifestovati. Leš mopsa starosti pet meseci donešen je zbog obdukcije na Katedru za sudsku veterinarsku medicinu, Fakulteta veterinarske medicine Univerziteta u Beogradu. Prvi klinički simptomi, u vidu opšte slabosti i poteškoća pri skakanju, javili su se kada je štene bilo staro tri i po meseca. Simptomi su u četvrtom mesecu postali mnogo ozbiljniji. Nativno rendgensko i mijelografsko snimanje pokazalo je uznapredovali stepen kifoze i kompresije kičmene moždine u T8-T9 regionu kičme. Nakon eutanazije, pas je obdukovan. Patomorfološki nalaz ukazao je na ozbiljnu deformaciju kičme, izraženu kifozu i klinast oblik T8 pršljena. Iako literaturni podaci ukazuju da klinički simptomi mogu biti nezapaženi sve do pet ili šest meseci starosti, ovaj slučaj pokazuje da čak kod mlade štenadi, starosti tri do četiri meseca, mogu biti prisutni ozbiljni simptomi. Ova činjenica je od velikog značaja, ne samo za veterinare, već i za odgajivače i vlasnike brahicefaličnih rasa pasa

Kliničko-epizootiološko ispitivanje piroplazmoze konja na prostoru Centralnog Balkana i njen uticaj na oštećenje DNK domaćina

Equine piroplasmosis is a significant vector-prone disease with a high degree of widespread and economic impact on horse industry throughout the world. The causative agents of equine piroplasmosis are protozoas Theileria equi and Babesia caballi. The disease can be present subclinical in horses, but also it can be accompanied by a serious clinical picture. The goal of this doctoral dissertation was the comprehensive clinical and epizootiological examination of the equine piroplasmosis on the central Balkan region, where we would investigate the presence of parasites by molecular detection methods, to follow the haematological and biochemical parameters, the parameters of oxidative stress and at the end, the degree of host cells DNA damage due to parasites presence. In order to gain insight into the prevalence of the disease, we conducted a molecular epidemiological study involving 165 horses, from eight locations from three countries (Serbia, Montenegro, Bosnia and Herzegovina). First, we did a PCR method for the detection of a 450 bp long segment of the 18S rRNA gene which is present in all Theileria and Babesia species. For the definitive species identification, a multiplex PCR was done. In order to confirm obtained results, sequencing of positive samples was made. The overall prevalence of T. equi and B. caballi in the study area was 32.1% and 1.8% respectively. In Serbia a prevalence of 40.2% was established for T. equi and 0.6% for B. caballi, in BiH 16.7% for T. equi and 4.2% for B. caballi, and in Montenegro 8.3% for T. equi and 8.3% for B. caballi. Results of the sequence analysis demonstrated 100% correspondence to multiplex PCR diagnosed Babesia species...Piroplazmoza konja predstavlja značajno vektorski prenosivo oboljenje sa velikom raširenošću i ekonomskim uticajem na konjarstvo širom sveta. Uzročnici piroplazmoze konja su protozoe Theileria equi i Babesia caballi. Oboljenje može biti prisutno kod konja i supklinički, ali može biti praćeno i ozbiljnom kliničkom slikom. Cilj ove doktorske disertacije je bio opsežno kliničko- epizootiološko ispitivanje priplazmoze konja na prostoru centralnog Balkana, gde bi ispitivali prisustvo parazita molekularnim metodama detekcije, pratili hematološke i biohemijske parametre, parametre oksidativnog stresa i na kraju stepen oštećenja DNK ćelija domaćina usled prisustva parazita. U cilju sticanja uvida u prevalenciju oboljenja sproveli smo molekularnu epizootiološku studiju koja je uključivala 165 konja, sa osam lokaliteta sa prostora tri države (Srbija, Crna Gora, Bosna i Hercegovina). Prvo smo uradili PCR metodu za detekciju 450 bp dugog segmenta 18S rRNK gena koji je prisutan kod svih Theileria i Babesia vrsta. Za definitivnu detekciju vrste je urađen multipleks PCR. Da bi potvrdili dobijene rezultate urađeno je sekvenciranje pozitivnih uzoraka. Ukupna prevalencija T. equi i B. caballi na ispitivanom prostoru iznosila je 32,1% i 1,8%, respektivno. U Srbiji je ustanovljena prevalencija od 40,2% za T. equi i 0,6% za B. caballi, u BiH 16,7% za T. equi i 4,2% za B. caballi, a u Crnoj Gori 8,3% za T. equi i 8,3% za B. caballi. Rezultati sekvenciranja su potvrdili 100% podudarnost sa vrstama ustanovljenim na osnovu metode multipleks PCR..

Clinical and epizootiological examination of equine piroplasmosis on the central balkan region and its effect on host DNA damage

Пироплазмоза коња представља значајно векторски преносиво обољење са великом раширеношћу и економским утицајем на коњарство широм света. Узрочници пироплазмозе коња су протозое Theileria equi и Babesia caballi. Обољење може бити присутно код коња и супклинички, али може бити праћено и озбиљном клиничком сликом. Циљ ове докторске дисертације је био опсежно клиничко- епизоотиолошко испитивање приплазмозе коња на простору централног Балкана, где би испитивали присуство паразита молекуларним методама детекције, пратили хематолошке и биохемијске параметре, параметре оксидативног стреса и на крају степен оштећења ДНК ћелија домаћина услед присуства паразита. У циљу стицања увида у преваленцију обољења спровели смо молекуларну епизоотиолошку студију која је укључивала 165 коња, са осам локалитета са простора три државе (Србија, Црна Гора, Босна и Херцеговина). Прво смо урадили PCR методу за детекцију 450 бп дугог сегмента 18S rRNК гена који је присутан код свих Theileria и Babesia врста. За дефинитивну детекцију врсте је урађен мултиплекс PCR. Да би потврдили добијене резултате урађено је секвенцирање позитивних узорака. Укупна преваленција T. equi и B. caballi на испитиваном простору износила је 32,1% и 1,8%, респективно. У Србији је установљена преваленција од 40,2% за Т. еqui и 0,6% за B. caballi, у БиХ 16,7% за Т. еqui и 4,2% за B. caballi, а у Црној Гори 8,3% за Т. еqui и 8,3% за B. caballi. Резултати секвенцирања су потврдили 100% подударност са врстама установљеним на основу методе мултиплекс PCR...Equine piroplasmosis is a significant vector-prone disease with a high degree of widespread and economic impact on horse industry throughout the world. The causative agents of equine piroplasmosis are protozoas Theileria equi and Babesia caballi. The disease can be present subclinical in horses, but also it can be accompanied by a serious clinical picture. The goal of this doctoral dissertation was the comprehensive clinical and epizootiological examination of the equine piroplasmosis on the central Balkan region, where we would investigate the presence of parasites by molecular detection methods, to follow the haematological and biochemical parameters, the parameters of oxidative stress and at the end, the degree of host cells DNA damage due to parasites presence. In order to gain insight into the prevalence of the disease, we conducted a molecular epidemiological study involving 165 horses, from eight locations from three countries (Serbia, Montenegro, Bosnia and Herzegovina). First, we did a PCR method for the detection of a 450 bp long segment of the 18S rRNA gene which is present in all Theileria and Babesia species. For the definitive species identification, a multiplex PCR was done. In order to confirm obtained results, sequencing of positive samples was made. The overall prevalence of T. equi and B. caballi in the study area was 32.1% and 1.8% respectively. In Serbia a prevalence of 40.2% was established for T. equi and 0.6% for B. caballi, in BiH 16.7% for T. equi and 4.2% for B. caballi, and in Montenegro 8.3% for T. equi and 8.3% for B. caballi. Results of the sequence analysis demonstrated 100% correspondence to multiplex PCR diagnosed Babesia species..

Crevna limfangiektazija kod pasa, teško do dijagnoze - 4 slučaja

Intestinal lymphangiectasia is an uncommon disease which can cause severe, chronic protein-losing enteropathy in dogs. Four dogs were presented at the Belgrade Clinic for Small Animals with clinical signs of chronic diarrhea, lethargy, anorexia, vomiting and weight loss. Abnormal physical examination findings included dehydration, signs of pain on abdominal palpation, and ascites. The most important clinicopathological findings were lymphopenia and hypoproteinemia with hypoalbuminemia. Abdominal ultrasound revealed intestinal abnormalities in all dogs. To establish an undoubted diagnosis of intestinal lymphangiectasia, endoscopy and histopathology were conducted.Crevna limfangiektazija predstavlja retko oboljenje kod pasa koje može izazvati ozbiljne, hronične enteropatije sa gubitkom proteina. Na Klinici za male životinje u Beogradu su primljena četiri psa sa simptomima hronične dijareje, letargije, anoreksije, povraćanja i gubitka na težini. Opštim pregledom su ustanovljene promene u smislu dehidratacije, bolnosti na palpaciju abdomena i ascitesa. Najznačajnije kliničko-patološke promene su bile limfopenija i hipoproteinemija sa hipoalbuminemijom. Ultrazvučnim pregledom abdomena su ustanovljene promene na crevima kod svih pasa. Da bismo postavili nesumljivu dijagnozu crevne limfangiektazije urađena je endoskopija i histopatologija

Improved DNA-based identification of cervidae species in forensic investigations

The main reasons for wildlife forensic research are animal poaching, illegal trade, and falsified game meat products. Small trace amounts, old and degraded materials present the most common samples in revealing criminal activities in this field. This is the reason why it is crucial to use adequate and reliable methods and samples to identify animal species killed outside the hunting season or species protected by law. In this study, different endpoint PCR and real-time PCR protocols were compared in the identification of three Cervidae species (Capreolus capreolus, Cervus elaphus, Dama dama) from old and damaged material found in an enclosed area where the animals were kept. From a total of 129 samples, end point PCR provided results for 119 samples, while real-time PCR was successful in all cases. Also, we created and tested a protocol for simultaneous analyses of different types of samples, which is of great importance as when the amplification is carried out simultaneously it is more cost efficient and speeds up the process

Efficacy of oral fluralaner for the treatment of canine generalized demodicosis: a molecular-level confirmation

BackgroundCanine generalized demodicosis is a common parasitic disease caused by the proliferation of Demodex mites. The introduction of isoxazoline class treatments in veterinary dermatology has resulted in apparently effective treatment of generalized demodicosis. The objective of this study was to evaluate the effectiveness of fluralaner for the treatment of canine generalized demodicosis using real-time PCR for the detection and quantification of Demodex DNA.MethodsTwenty privately owned dogs with clinical symptoms of generalized demodicosis and deep skin scrapings positive for Demodex canis mites were enrolled in the study. Following diagnosis (day 0) each dog was treated with fluralaner at the recommended commercial dose for tick and flea treatment (25-56 mg/kg) based on body weight. Clinical and mite count assessments, and hair sampling for molecular analyses were performed on days 0, 28, 56, 84 and 112. Demodex DNA was detected and quantified using real-time PCR.ResultsA single oral dose of fluralaner reduced Demodex mite counts in skin scrapings by an average of 98.9% in all dogs by day 28. No mites were recovered from skin scrapings from any treated dog by day 56, at which time the dog was considered to be clinically cured, with total hair regrowth. There were significant differences among examined dogs in qPCR cycle threshold (Ct) values on days 0, 28, 56, 84 and 112. Demodex DNA levels decreased (increasing Ct values) throughout the study. Mite DNA was present on day 112, possibly from dead mites, at values significantly lower than in samples taken on days 0, 28 and 56. Based on qPCR testing of diluted samples, the Demodex mite population was reduced by approximately 1000-fold on day 112.ConclusionsOral administration of fluralaner at the recommended dose to dogs with generalized demodicosis is highly effective for reducing Demodex mite populations and resolving clinical signs of generalized demodicosis. The presence of mite DNA may indicate that treatment did not kill all Demodex mites

Lafora disease in a Chihuahua dog: a case report

The aging process is still not fully understood, although it has been studied for centuries. One of the processes in the brain during aging is the accumulation of extracellular and intracellular deposits of amyloid and lipofuscin. Deposits of various polyglucosan bodies (PGBs) are also found in brain tissue. The accumulation of the Lafora bodies (LB), a type of PGBs, can cause the Lafora disease (LD). Initial signs of the disease in humans are tonic-clonic seizures with blindness and myoclonus seizures. Normally, all haematological and biochemical indices are within the reference range in dogs with this disease. In this case, a 7-year-old Chihuahua dog with tonic-clonic seizures was presented. According to history, neurological examination, and blood test, toxic, metabolic, and infectious causes for the seizures were ruled out. The dog was started on phenobarbital 2.5 mg/kg twice daily per os. Two years later, the dog died due to complications caused by a duodenal foreign body. Postmortem examination revealed hyperaemic meninges and an enlarged, oedematous brain with flattened gyri and narrowed sulci. Histopathological examination revealed multifocal to diffuse, randomly distributed PGBs that were positive on periodic acid-Schiff (PAS) staining. The diagnosis of LD in dogs is of great importance because they are a good experimental model for neurological studies of neurodegenerative diseases in humans

Ultrazvučni pregled abdomena pasa

Ultrazvučni pregled, poznat i kao ultrasonografija, je neinvaziv- na tehnika snimanja koja omogućava uočavanje unutrašnjih telesnih struktura beleženjem odjeka ili refleksije ultrazvučnih talasa. Za razli- ku od rendgenskih zraka koji su potencijalno opasni, ultrazvučni talasi se smatraju sigurnim. Svaki veterinar kliničar treba da bude svestan mogućnosti, ali i ograničenja ove dijagnostičke procedure. On mora da zna na koja pitanja ovaj vid pregleda može dati odgovore, ali i da bude svestan činjenice da negativan ultrazvučni nalaz ne isključuje uvek oboljenje. Najbitniji segment svakog ultrazvučnog pregleda je sistematičnost. Kod svakog pacijenta se treba držati istog redosleda pregleda organa. Ultrazvučni pregled abdomena psa obuhvata pre- gled hepatobilijarnog sistema, želuca, slezine, bubrega, nadbubrež- nih žlezdi, mokraćne bešike, genitalnih organa, creva, limfnih čvorova i krvnih sudova. Standardizovano, precizno i konkretno pisanje izve- štaja, nakon obavljenog ultrazvučnog pregleda, treba da bude velika pomoć veterinaru pri postavljanju dijagnoze ili planiranju sledećih di- jagnostičkih procedura

Molekularna detekcija Babesia spp. u krpeljima uzorkovanim sa asimptomatskih pasa na području određenih beogradskih opština

Babesiosis of domestic animals is a vector transmissible and clinically significant disease, caused by protozoa of genus Babesia and Theileria. Possible causative agents for this disease in dogs in Europe are: Babesia canis, B. gibsoni, B. vogeli and B. microti-like. Diagnostics of babesiosis of dogs was for a long time based on the visual inspection of stained blood smear under a microscope, while today there have been increasingly used molecular methods of detection in precise, species diagnostics. The objective of this work was molecular detection of the cause of babesiosis of dogs in the ticks sampled from asymptomatic dogs in the region of some Belgrade municipalities, all for better understanding of epizootiological situation. From three sites in Belgrade, there were collected 49 ticks, sampled from the dogs with no symptoms. There was carried out the determination of the ticks, and after that, DNA was isolated for molecular examination. First, theret was performed Polymerase Chain Reaction (PCR), for determining the species of the genus Babesia, and after that there was also carried out the determining of polymorphism in the length of restriction fragments (RFLP) for the purpose of the causative agent species determination. Out of the total number of the examined ticks, 18,34% were positive on Babesia spp. By RFLP method, in two cases (4,08%) B. Gibsoni was identified, while in 7 cases (14,92%) there were no restriction sites for the used enzymes, what suggests that most likely it was B. canis. The ticks positive on the cause of babesiosis were: Dermacentor reticulatus (4 cases), Rhipicephalus sanguineus (4 cases) i Ixodes ricinus (1 case). This work confirms the presence of Babesia spp. in the ticks sampled from asmptomatic dogs on the territory of Belgrade as well as the significance of PCR-RFLP method in diagnostics and identification of the causative agent of babesiosis in dogs. For the first time in Serbia, there was determined the presence of B. gibsoni in ticks (Species Rhipicephalus sanguineus).Babezioza domaćih životinja predstavlja vektorski prenosivo i klinički veoma značajno oboljenje, uzrokovano protozoama rodova Babesia i Theileria. Uzročnici koji mogu da izazovu oboljenje kod pasa u Evropi su: Babesia canis, B. gibsoni, B. vogeli i B. microti-like. Dijagnostika babezioze pasa se dugo vremena bazirala na vizuelizaciji uzročnika pregledom obojenog krvnog razmaza pod mikroskopom, dok se danas sve više koriste molekularne metode detekcije u preciznoj, specijskoj dijagnostici. Cilj ovog rada je bila molekularna detekcija različitih uzročnika babezioze pasa u krpeljima uzorkovanim sa asimptomatskih pasa na prostoru određenih beogradskih opština, radi boljeg razumevanja epizootiološke situacije. Sa tri lokacije u Beogradu je prikupljeno 49 krpelja, uzorkovanih sa pasa bez simptoma bolesti. Izvršena je determinacja krpelja, a nakon toga je izolovana DNK za molekularna ispitivanja. Prvo je urađena reakcija lančane polimerizacije (PCR) za utvrđivanje vrsta iz roda Babesia, a nakon toga i određivanje polimorfizma u dužini restrikcionih fragmenata (RFLP) u cilju specijske identifikacije uzročnika. Od ukupnog broja ispitanih krpelja 18,34% je bilo pozitivno na Babesia spp. RFLP metodom je u dva slučaja (4,08%) identifikovana B. gibsoni, dok u 7 slučajeva (14,92%) nije bilo restrkicionih mesta za korišćene enzime, što ukazuje da se najverovatnije radilo o B. canis. Pozitivni krpelji na uzročnike babezioze su bili Dermacentor reticulatus (4 slučaja), Rhipicephalus sanguineus (4 slučaja) i Ixodes ricinus (1 slučaj). Ovaj rad potvrđuje prisustvo Babesia spp. u krpeljima uzorkovanim sa asimptomatskih pasa na teritoriji grada Beograda, kao i značaj PCR-RFLP metode u dijagnostici i identifikaciji uzročnika babezioze pasa. Prvi put u Srbiji je utvrđeno prisustvo B. gibsoni kod krpelja (vrsta Rhipicephalus sanguineus)

A molecular and haematological study of Theileria equi in Balkan donkeys

Equine piroplasmosis in donkeys has been recognised as a serious problem of major economic importance. The present molecular study is the first investigation of the presence of Theileria equi and Babesia caballi in Balkan donkeys and of the possible haematological alterations related to it. A total of 70 apparently healthy donkeys from Serbia were included in this study. The overall prevalence of T. equi infection in donkeys tested with multiplex PCR was 50%. There was no B. caballi-positive sample. Infections in donkeys included in this study seem to be associated with decreased red blood cell count, haemoglobin concentration, haematocrit and platelet count, and with increased white blood cell count, mean corpuscular haemoglobin and mean corpuscular haemoglobin concentration. Altered haematological parameters in donkeys can lead to a decrease in working capacity and production performance. Further molecular research and long-term monitoring of equine piroplasmosis is needed in Serbia and throughout Europe