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

Characterization of sex - specific DNA sequences for the species and sex determination of protected birds for forensic examinations

Најзначајније претње које утичу на смањење бројности многих угрожених врста јесу криволов, илегалан лов, као и незаконита трговина и транспорт дивљих животиња. Када је у питању економски аспект, ове криминалне радње уједно представљају једну од најпрофитабилнијих илегалних трговина на свету. ДНК анализе из области форензичких испитивања дивљих животиња омогућавају идентификацију угрожених животињских и биљних врста које су предмет илегалне трговине. Због недвосмислене идентификације и могућности да се користе веома мали (па чак и деградирани) биолошки узорци, из различитих извора (нпр. ткива, кости, перје, љуске јаја, длаке, крљушти), молекуларно-генетичке технике постају све важније у откривању илегалног лова и промета дивљих животиња. Ова релативно нова дисциплина сада постаје кључно средство за борбу против илегалне трговине животињама и помоћ агенцијама за спровођење закона и управљање природним ресурсима. Циљ ове докторске дисертације je развијање молекуларно-генетичке методе за истовремено одређивање врсте и пола птица анализом CHD гена, затим дизајнирање специфичних прајмера за одабране врсте, као и њихова провера и могућност коришћења. Као материјал за ово истраживање коришћени су узорци перја пореклом од птица из Београдског зоолошког врта и узорака угинулих заштићених врста птица које су обдуковане на Катедри за судску ветеринарску медицину и законске прописе Факултета ветеринарске медицине, Универзитета у Београду (ФВМ УБ). Додатно, коришћени су и узорци из архиве Катедре за биологију ФВМ УБ. Кoришћeњeм Polymerase Chain Reaction (PCR) тeхникe и сета прајмера обележених флуоресцентним бојама CHD1F/CHD1R и P1/P8 вршена је амплификација дела Chromo Helicase DNAbinding (CHD1) гена и визуелизација продуката хоризонталном електрофорезом на агарозном гелу и методом капиларне електрофорезе. За добијање ампликона за секвенцирање, у циљу креирања нових прајмера за одређивање пола одређених врста, коришћен је необележени сет прајмера 2550F/2718R и P2/P8. Такође, извршено је и секвенцирање амплификата девет врста птица, у циљу добијања потпунијих информација о њима. Након секвенцирања и анализе добијених секвенци, креиран је нови пар прајмера за врсту Cacatua alba. Taкође, добијене секвенце су депоноване у online генску базу (NCBI, https://www.ncbi.nlm.nih.gov). Одређивање пола методом гел електрофорезе било је успешно код 60 јединки птица од укупно 71 испитиваног узорка. За птице из реда Strigiformes и птице из фамилије Fringilidae добијени резултати нису били поуздани, односно, производи амплификације добијени хоризонталном електрофорезом на агарозном гелу нису били јасни да би могли бити сигурни да ли су испитиване јединке мужјаци или женке. Такође, методом гел електрофорезе није било могуће утврдити прецизну величину CHD1 ампликона. За разлику од гел електрофорезе, капиларна електрофореза се показала успешном код свих испитиваних птица када је у питању детерминација пола, јер је омогућила добијање недвосмислених резултата код свих анализираних узорака. Када је у питању идентификација врсте коришћењем капиларне електрофорезе, ова метода се такође показала врло успешном. Код свих испитиваних јединки одређене су величине фрагмента CHD гена на W и Z хромозомима које служе при идентификацији врсте. Добијени резултати указују да се капиларна електрофореза може користити као врло поуздана, брза и исплатива метода у циљу идентификације како врсте, тако и пола птица. Гел електрофореза је врло корисна за одређивање пола, али за све узорке код којих пол није могао бити детерминисан овом методом, капиларна електрофореза је била успешна. Секвенцирањe обезбеђује утврђивање односа између врста и креирање нових и поузданијих прајмера. Применом наведених метода могуће је из остатака ткива утврдити врсту птице која је предмет форензичке анализе и потом на основу законске регулативе одредити степен њене заштите. На овај начин доприноси се истрази у спровођењу поступка против починиоца кривичног дела илегалног транспорта дивљих и угрожених птица. Добијени резултати су од великог значаја и за даљи развој ових метода у функцији ветеринарске форензике и за смањење стопе илегалне трговине животињама у Републици Србије, али и у целом свету

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

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)

Molekularna detekcija i ispitivanje zastupljenosti uzročnika Dirofilarioze pasa na teritoriji grada Beograda

Dirofilariosis in dogs is a parasitic disease caused by the strains Dirofilaria immitis and D. Repens. Dirofilaria immitis parasitizes in the right side of the heart and pulmonary artery, causing considerable functional impairment of the heart, while adult forms of D. repens mostly parasitize in subcutaneous tissue causing minor skin damage. In this investigation, during the period from May to November 2015, 60 dogs of different race from the territory of Belgrade were examined. Ater the clinical examination, the blood was sampled for microfilariae detection by Knott test, as well as for molecular detection of the agent. Main clinical signs were: fatigue during exersise, shortness of breath and dry cough, anorexia, rapid heartbeat and skin changes. Microfilariae were detected in 22 dogs (37%) by modified Knott test. By molecular genetic method there was determined the prevalence of D. immitis 42%, and D. repens 22%. In this investigation 7 dogs were negative on Knott test, but by PCR method they showed a positive reaction on the presence of species of the genus Dirofilaria. On the basis of the obtained results, the application of molecular-genetic methods is justified in accurate and reliable species identification of Dirofilaria genus.Dirofilarioza pasa je parazitsko oboljenje izazvano vrstama Dirofilaria immitis i D. repens. D. immitis parazitira u desnoj strani srca i plućnoj arteriji, dovodeći do značajnog funkcionalnog oštećenja srca, dok adultni oblici D. repens uglavnom parazitiraju u subkutanom tkivu dovodeći do manjih oštećenja kože. U ovom istraživanju ispitano je 60 pasa, različitih rasa, sa teritorije grada Beograda, u periodu od maja do novembra 2015. godine. Nakon kliničkog pregleda, krv je uzorkovana za detekciju mikrofilarija modifikovanim Knott-ovim testom, kao i za molekularnu detekciju uzročnika. Glavni klinički znaci su bili: brzo zamaranje tokom treninga, otežano disanje i suv kašalj, anoreksija, ubrzan rad srca i promene na koži. Mikrofilarije su detektovane kod 22 psa (37%) modifikovanim Knott-ovim testom. Molekularno genetskom metodom utvrđena je prevalencija D.immitis 42%, a D. repens 22 %. U sprovedenom istraživanju je bilo 7 pasa koji su bili negativni na Knott testu, dok su metodom PCR pokazali pozitivnu reakciju na prisustvo vrsta iz roda Dirofilaria. Na osnovu dobijenih rezultata primena molekularno-genetskih metoda potvrđuje svoju opravdanost u preciznoj i pouzdanoj specijskoj identifikaciji roda Dirofilaria

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

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

First report of Polycystic kidney disease occurrence in Persian cats in Serbia

Polycystic kidney disease (PKD) is an inherited autosomal disorder in cats, mostly diagnosed in Persian cats. Renal cysts can be diagnosed by ultrasound, but cats must be at least 16 weeks old. The goals of this study were to assess the occurrence of PKD in Serbia using a randomly selected group of Persian cats, to compare the diagnostic efficacy of ultrasound and genetic tests, and to measure haematological and selected biochemical parameters. We examined 70 cats of Persian breed, between 4 months and 8 years of age. Complete blood count and selected biochemical parameters were measured, renal ultrasound was performed. Swabs of the oral cavity were obtained for genetic testing. Percentage of PKD positive cats identified by genetic testing was 48.6%, whilst only 18.6% were detected through ultrasound. Animals that were polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) positive and ultrasound negative ranged from 4 months to 3.5 years. All haematological and biochemical parameters were within the the normal range values in all examined cats. Genetic methods proved to be the most effective for reliable and early diagnosis of PKD in Persian cats. DNA analysis can be used right after birth, and excludes the need for other diagnostic procedures, such as ultrasound