ISSN 1580-4003 THE SCIENTIFIC JOURNAL OF THE VETERINARY FACULTY UNIVERSITY OF LJUBLJANA SLOVENIAN VETERINARY RESEARCH SLOVENSKI VETERINARSKI ZBORNIK Volume > 44 4 Slov Vet Res • Ljubljana • 2007 • Volume 44 • Number 4 • 97-137 THE SCIENTIFIC JOURNAL OF THE VETERINARY FACULTY UNIVERSITY OF LJUBLJANA SLOVENIAN VETERINARY RESEARCH SLOVENSKI VETERINARSKI ZBORNIK Volume / 44 4 Slov Vet Res • Ljubljana • 2007 • Volume 44 • Number 4 • 97-137 The Scientific Journal of the Veterinary Faculty University of Ljubljana SLOVENIAN VETERINARY RESEARCH SLOVENSKI VETERINARSKI ZBORNIK Previously: RESEARCH REPORTS OF THE VETERINARY FACULTY UNIVERSITY OF LJUBLJANA Prej: ZBORNIK VETERINARSKE FAKULTETE UNIVERZA V LJUBLJANI 4 issues per year / izhaja štirikrat letno Editor in Chief / glavni in odgovorni urednik: Gregor Majdič Technical Editor / tehnični urednik: Matjaž Uršič Assistant to Editor / pomočnica urednika: Malan Štrbenc Editorial Board / Uredniški Odbor: Vesna Cerkvenik Flajs, Vojteh Cestnik, Polona Juntes, Matjaž Ocepek, Zlatko Pavlica, Uroš Pestevšek, Modest Vengušt, Milka Vrecl, Olga Zorman Rojs, Veterinary Faculty University of Ljubljana / Veterinarska fakulteta Univerze v Ljubljani Editorial Advisers / svetovalca uredniškega odbora: Gita Grecs-Smole for bibliography (bibliotekarka), Leon Ščuka for statistics (za statistiko) Reviewing Editorial Board / ocenjevalni uredniški odbor: Ivor D. Bowen, Cardiff School of Biosciences, Cardiff, Wales, UK; Rudolf Cabadaj, University of Veterinary Medicine, Košice, Slovakia; Antonio Cruz, Departement of Clinical Studies, Ontario Veterinary College, Guelph, Ontario, Kanada; Gerry M. Dorrestein, Duch Research Institute for Birds and Exotic Animals, Veldhoven, The Netherlands; Wolfgang Henninger, Veterinärmedizinische Universität Wien, Austria; Simon Horvat, Biotehniška fakulteta, Univerza v Ljubljani, Slovenia; Josef Leibetseder, Veterinärmedizinische Universität Wien, Austria; Louis Lefaucheur, INRA, Saint-Gilles, France; Bela Nagy, Veterinary Medical Research Institute Budapest, Hungary; Peter O'Shaughnessy, Institute of Comparative Medicine, Faculty of Veterinary Medicine, University of Glasgow, Scotland, UK; Milan Pogačnik, Veterinarska fakulteta, Univerza v Ljubljani, Slovenia; Peter Popelka, University of Veterinary Medicine, Košice, Slovakia; Detlef Rath, Institut für Tierzucht, Forschungsbericht Biotechnologie, Bundesforschungsanstalt für Landwirtschaft (FAL), Neustadt, Germany; HansPeter Sallmann, Tierärtzliche Hochschule Hannover, Germany; Marko Tadic, Veterinarski fakultet, Sveučilište u Zagrebu, Croatia; Frank J. M. Verstraete, University of California Davis, Davis, California, US Slovenian Language Revision / lektor za slovenski jezik: Viktor Majdič Address: Veterinary Faculty, Gerbičeva 60, 1000 Ljubljana, Slovenia Naslov: Veterinarska fakulteta, Gerbičeva 60, 1000 Ljubljana, Slovenija Tel.: +386 (0)1 47 79 100, 47 79 129, Fax: +386 (0)1 28 32 243 E-mail: slovetres@vf.uni-lj.si Sponsored by the Slovenian Research Agency Sofinancira: Agencija za raziskovalno dejavnost Republike Slovenije ISSN 1580-4003 Printed by / tisk: Birografika Bori d.o.o., Ljubljana Indexed in / indeksirano v: Agris, Biomedicina Slovenica, CAB Abstracts, IVSI Urlich's International Periodicals Directory http://www.vf.uni-lj.si/veterina/zbornik.htm SLOVENIAN VETERINARY RESEARCH SLOVENSKI VETERINARSKI ZBORNIK Slov Vet Res 2007; 44 (4) Letter to Editor Useh NM. Constraints of blackleg control in Nigeria................................................101 Case Reports Nemec A, Pavlica Z, Juntes P, Crossley DA. Advanced periodontal disease in a Yorkshire terrier with concurrent nasal cavity malignancy.................................................................103 Plavec T, Tozon N, Kotnik T. Generalized symmetric alopecia and hyperoestrogenism associated with concurrent lymphoma, sertoli cell tumour and seminoma in a Samoyed ......................................109 Seliskar A, Zdovc I, Zorko B. Nosocomial Klebsiella oxytoca infection in two dogs.........................115 Kataria AK, Kataria N, Gahlot AK. Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert of India....................................................................123 Cestnik V. Prof. dr. Milan Pogacnik - Doctor honoris causa..........................................133 Subject Index Volume 44, 2007............................................................... 135 Author Index Volume 44, 2007 ............................................................... 137 Slov Vet Res 2007; 44 (4): 101-2 UDC 636.2.09:579.852.11:616-036.22:615.83(669) Letter to Editor CONSTRAINTS OF BLACKLEG CONTROL IN NIGERIA N. M. Useh Department of Veterinary Pathology and Microbiology, Faculty of Veterinary Medicine, Ahmadu Bello University, Zaria, Nigeria E-mail: nickuseh@yahoo.com Sir, blackleg (also known as symptomatic anthrax, blackquarter or emphysematous gangrene) is a disease of cattle, sheep and other ruminants (1). In Nigeria, the disease was first reported in 1929 and has remained a major problem of cattle in the country (2). Infective spores of Clostridium chauvoei ingested during grazing are lodged in the gastrointestinal tract (GIT), livers and spleens of healthy cattle (3) and remain latent until their germination is triggered by punctured wounds (4). During growth, the organism is known to produce neuraminidase (sialidase), an enzyme that plays a key role in the pathogenesis of blackleg (5). Although Agba and Princewill (1986) (6) put the economic losses of cattle due to blackleg in Nigeria at less than a million United States Dollars (USD) annually, the current losses to the disease may approximate about 4.3 million USD annually (7). This is because of increased annual outbreaks over the years and the deregulation of Nigeria's economy. This letter to the Editor highlights the constraints associated with the control of blackleg of cattle in Nigeria and the possible ways of ameliorating the constraints. The nomadic Fulani pastoralists of rural Nigeria, who own about 70-80% of livestock in the country, migrate from one place to another in search of grazing pasture for their livestock (8). It therefore follows that as they migrate, they encounter soils with high proportion of clostridial spores that constitute a health hazard. It is common knowledge that the best control strategy against blackleg is vaccination (9). Most times, potent vaccines are difficult to come by in Nigeria, because of the inability to maintain the cold chain. The nomads a times purchase the vaccines on their own from veterinary shops, without any machinery in place to maintain the cold chain. This practice has made it difficult to effectively control the disease in Nigeria. In line with this, therefore, some state governments in Nigeria do not vaccinate animals routinely against blackleg, because the no- mads do not request for it, except in times of disease outbreaks. Even in the face of outbreaks, the attitude of the nomads in the control of disease spread to neighbouring herds leaves much to be desired. They do not report the outbreaks and may chose to move away from the area. The drug of choice for treating blackleg is penicillin (7) but the nomads prefer the use of herbal remedies to treat the disease. They may report outbreaks to veterinarians and government officials only if the herbal remedies do not achieve the desired therapeutic results, accompanied by an upsurge of mortality which they can not control. Two herbal remedies (Tamarindus indicus and Combretumfragrans) are preferred to penicillin by the nomads for treating blackleg (10). The side effects of herbal preparations have been identified and they include: inappropriate dosing (11), intoxication leading to death of treated animals (12) or the problem of partial efficacy associated with some herbal remedies (13). The control of blackleg has remained a major problem in Nigeria, since the nomads of rural Nigeria who are key players in the livestock industry are not settled and continue to move from one Clostrid-ium infected soil to the other in search of grazing pasture. It is recommended that animal ranches (settled farms) should be established in areas free of clostridial spores. This is because of the danger posed by this on the health of animals and the role of the spores in the pathogenesis of blackleg. To effectively control the disease in cattle, vaccination of the animals using potent vaccines has been advocated. It is concluded that, while government should be prevailed upon to revive the available grazing reserves to settle the Fulanis, there is the need to encourage the use of the herbal remedies, as they are cheaper, effective and available in Nigeria. Research should be conducted to establish the dosage regiments, therapeutic index and side effects of these herbs. 102 N. M. Useh References 1. Armstrong HL, MacNamee JK. Blackquarter in deer. J Am Vet Med Assoc 1950; 117: 212-4. 2. Osiyemi T I O. The aetiology and data on seasonal incidence of clinical blackleg in Nigerian cattle. Bull Anim Health Prod Africa 1975; 23: 367-70. 3. Kerry JB. A note on the occurrence of Clostridium chauvoei in the spleens and livers of normal cattle. Vet Rec1964; 76: 396. 4. Mohammed OE, Tageldin MH, El-Sanousi SM. Some observations on the pathogenicity of blackleg. Bull Anim Hlth Prod Afri 1990; 38: 355. 5. Useh NM , Nok AJ, Ajanusi OJ, Balogun EO, Oladele SB, Esievo KAN. In vitro production of neuraminidase by Clostridium chauvoei (Jakari strain). Vet Arh 2004; 74: 289-98. 6. Agba MI, Princewill TJT. Combined clostridial vaccine for controlling blackquarter in Nigeria. In: XIV International Congress of Microbiology. Manchester, 1986: 120-1. 7. Useh N M. The possible role of clostridial neuramini-dase in the pathogenesis of blackleg in Zebu cattle. Zaria, Nigeria : Ahmadu Bello University, 2006: 172 str. Ph. D. Dissertation. 8. Suleiman H. Policy issues on pastoral development. In: Pastoralism in Nigeria: past, present and future. Proceedings of the National Conference on Pastoralism, Lagos, Nigeria, 1988. 9. Kijima-Tanaka M, Ogikubo Y, Kijima A, Sasaki Y. Flagella based enzyme-linked immunosorbent assay for evaluation of the immunity in mice vaccinated with blackleg vaccine. J Microbiol Meth 1998; 32: 79-85. 10. Abdu PA, Jagun AG, Gefu JO, Mohammed AK, Al-awa CBI, Omokanye AT. A survey of ethnoveterinary practices of agropastoralists in Nigeria: In: Gefu JO, Abdu PA, Alawa CB, eds. Ethnoveterinary practices: research and development. Zaria: National Animal Production Research Institute; Ahmadu Bello University, Nigeria, 2000. 11. Gueye E. Ethnoveterinary medicine against poultry diseases in African villages. World Poultr Sci J 1999; 55: 187-98. 12. Nomoko M. Cassia sieberiana De (Caesalpiniace'es). Le Flamboyant 1997; 43: 4. 13. Sonaiya E B. The context and development of small holder rural poultry production in Africa. In: Proceedins of CTA seminar on smallholder rural poultry production. Thessaloniki, Greece, 1990: Vol. 1: 35-52. Slov Vet Res 2007; 44 (4): 103-8 UDC 636.7.09:616.31-006 Case Report ADVANCED PERIODONTAL DISEASE IN A YORKSHIRE TERRIER WITH CONCURRENT NASAL CAVITY MALIGNANCY Ana Nemec1*, Zlatko Pavlica1, Polona Juntes2, David A. Crossley1 1 Clinic for Surgery and Small Animal Medicine, 2Institute of Pathology, Forensic and Administrative Veterinary Medicine, Veterinary Faculty, Gerbiceva 60, 1000 Ljubljana, Slovenia Corresponding author, E-mail: ana.nemec@vf.uni-lj.si Summary: An eleven-year-old male Yorkshire terrier weighing 2.6 kg with a primarily indoor lifestyle was presented for cardiopulmonary examination due to a 6-month history of difficulty breathing. No cardiopulmonary abnormalities were detected but the history included bilateral serous nasal discharge and oral abnormalities were evident. Examination under general anaesthesia confirmed advanced periodontal disease with oronasal fistulae detected at the maxillary canine teeth. Dental treatment, including repair of the oronasal fistulas, appeared to resolve the respiratory signs but the discharge reappeared at the left nostril 1 month later. There was no evidence of persistent or recurrent oronasal fistula, so rhinoscopy was performed identifying a mass in the left nasal cavity. Histopathological examination identified the biopsy specimen as a low-grade predominantly papillary-cystic adenocarcinoma combined with transitional carcinoma. Key words: periodontal diseases; rhinitis; nose neoplasms - pathology; adenocarcinoma; dogs Introduction Nasal discharge is a common problem in dogs (1). It may be the result of nasal or paranasal disorders or be related to systemic disease (1). In older animals chronic nasal discharge is commonly due to extension of dental disease to involve the nasal cavities (oronasal fistula or tooth root abscessation) or neoplasia (1, 2). Other differentials include fungal infection, chronic foreign bodies, allergic and nonspecific rhinitis as well as systemic disease (1, 3). Both fungal rhinitis and nasal foreign bodies tend to be seen in dogs that spend a lot of time outdoors (1). Except for viral infections and systemic diseases, the nasal discharge in animals with nasal conditions is usually unilateral or may become bilateral with disease progression (1). The case presented here illustrates some of the difficulties in diagnosis and treatment of nasal conditions in dogs. Received: 3 July 2007 Accepted for publication: 17 August 2007 Clinical case Presentation An eleven-year-old male Yorkshire terrier weighing 2.6 kg with a primarily indoor lifestyle was referred to the Clinic for Surgery and Small Animal Medicine of the Veterinary Faculty in Ljubljana for cardiopulmonary examination in January 2007. On presentation at the clinic the dog had a six month history of difficulty breathing, especially during the night and the owner mentioned a serous nasal discharge, more pronounced from the left nostril. The dog had been treated with furosemide (Edemid; Lek Ljubljana, Slovenia; 1mg/kg/day p.o.) and ramipril (Tritace; Aventis Pharma, Austria; 0.5 mg/kg/day p.o.) for one month prior to referral. No other problems were reported in the history. Complete blood count (CBC) values were within normal limits. On auscultation no cardiac abnormalities were detected, lung sounds and pulse were normal, but there was a pronounced expiratory stertor due to partial nasal airflow obstruction. Oral examination revealed the presence of advanced periodontal disease. As the dog's problem appeared to be related to the upper airway, cardiopulmonary examinations were post- 104 A. Nemec, Z. Pavlica, P. Juntes, D. A. Crossley poned and the previously prescribed cardiologic treatment was discontinued pending the results of upper aerodigestive tract examination. The dog was scheduled for general anaesthesia the next day to permit a more thorough oral examination and treatment of the dental disease. Anaesthesia The dog was premedicated with methadone (Hep-tanon; Pliva, Croatia; 0.4 mg/kg i.m.) and carprofen (Rimadyl; Pfizer Animal Health S.A., UK; 4 mg/kg i.v.) prior to induction of anaesthesia using propofol (Propofol 1% Fresenius; Fresenius Kabi, Austria; 7.5 mg/kg i.v.). Following endotracheal intubation, anaesthesia was maintained with isoflurane (Forane; Abbott Laboratories Ltd., GB) given to effect (approximately 1.5%) in oxygen (2 l/min) using a Mapleson F anaesthetic circuit. Amoxicillin and clavulanic acid (Synulox; Pfizer Italia S.r.l., Italia; 20 mg/kg s.c.) was administered preoperatively as the start of a 10 day course of treatment (20 mg/kg/12h p.o.), carprofen treatment also being continued for 5 days (4 mg/ kg/day p.o.) to maintain analgesia during the postoperative period. During anaesthesia body temperature, respiratory rate, inspired and expired isoflu-rane, heart rate, ECG, pulse oximetry, end tidal CO2 and blood pressure with Doppler manometer were monitored. During the procedure and recovery from anaesthesia fluid homeostasis was maintained by administration of Ringer's lactate solution 26 ml/h (Hartman's solution; B.Braun Melsungen AG; Germany) i.v. Oralfindings The dog's oral cavity was assessed by means of periodontal examination and recording, plus radiography of disease affected areas. Tooth presence, probing depth, periodontal attachment loss, furcation involvement and tooth mobility were graded. Supra- and subgingival scaling were then performed, followed by polishing and gingival lavage with water, prior to extraction of compromised teeth. Oral examination revealed advanced periodontal disease with generalised plaque and calculus accumulation. Many teeth were already missing. Oronasal fistulae were detected palatal to both maxillary canine teeth with deep periodontal pockets being present buccally; probing depths were 10 mm on the left and more than 12 mm on the right. Generalised gingival recession of about 2 mm was detected affecting those mandibular premolar and molar teeth that were still present. Of the remaining incisor teeth, only the left maxillary third incisor tooth was stable, the rest being highly mobile. There was generalized bleeding on periodontal probing, due to the extent of gingivitis and periodontitis. Dental treatment Left and right infraorbital and mental nerve blocks were performed using bupivacaine (Mar-caine 0.5%; AstraZeneca, UK; 0.05 ml per site). Gross deposits were removed from the teeth and the oral cavity rinsed thoroughly prior to extraction of compromised teeth. Extraction was performed after sectioning of multirooted teeth (using cutting burs in a high-speed dental handpiece with copious water spray). A combination of closed elevation and luxation of mobile single-rooted teeth/tooth segments, and open extraction technique, raising muco-gingival access flaps with or without alveolar bone removal as required to facilitate elevation/luxation of the remaining teeth/roots. All the right maxillary incisor teeth, the canine, the right maxillary first, second and third premolar teeth, the left maxillary second incisor tooth, all the mandibular incisor teeth and the right mandibular canine tooth were extracted. There was extensive bleeding while raising mucogingival access flaps in the severely inflamed gingival tissues leading to significant blood loss (estimated from the number of swabs used and their degree of blood saturation to about 40 ml) resulting in a blood pressure drop which required use of a plasma expander (6% HES; Fresenius Kabi Deutschland GmbH, Germany; 10 ml bolus and Hartman's solution 50 ml/kg/h). In view of this, and a drop in the body temperature (to 35 °C), it was decided not to complete the treatment in a single session but to stage it with the aim of performing extractions of the remaining compromised teeth at a later date. Second presentation and dental treatment One and a half months after the initial treatment the dog was generally well, however, there was still a serous nasal discharge from the left nostril and an oronasal fistula was still present palatal to the left maxillary canine tooth. All CBC values were still within normal limits so, two months after the first treatment (March 2007), the dog was re-examined under general anaesthesia, using the same protocol as previously. Oral examination revealed severe plaque accumulation, however, gingival recession previously affecting the mandibular premolar and molar teeth has healed only after scaling and polishing, as had the mucogingival access flaps, al- Advanced periodontal disease in a Yorkshire terrier with concurrent nasal cavity malignancy 105 though the monofilament resorbable suture material (Biosyn 5-0; United States Surgical, USA) was still present. After thorough oral cleaning (scaling and polishing), the left maxillary canine tooth, left mandibular canine tooth and left mandibular first premolar tooth were extracted using the techniques described previously, the oronasal fistula being closed with a single layer mucogingival flap. A thorough examination of the larynx, pharynx, soft palate and caudal nasal cavity using a dental speculum, mirror and retractor was performed, but no additional abnormalities were found. Due to the presence of the oronasal fistula the dog was maintained on amoxicillin and clavulanic acid (20 mg/ kg/12 hours p.o.) for 10 days following the dental treatment, with carprofen (4 mg/kg/day) also being given for the first 4 days. Further presentations and diagnostic procedures One month after the second dental treatment (April 2007) the owner reported reappearance of the serous nasal discharge from the left nostril, it having stopped shortly after the previous treatment, and mild difficulties breathing. As no recurrence of the oronasal fistula was detected on clinical examination, the owner agreed to have another examination under general anaesthesia, but the owner scheduled this for 1 month later (May 2007). At this time all CBC values, urea, creatinine, alkaline phosphatase and alanine aminotransferase were still within normal limits. Examination under general anaesthesia (induced and maintained as previously, but without antibiotics and carprofen) revealed no abnormalities in the oral cavity. Radiographs of the head (lateral, open-mouth and intra-oral occlusal dorsoventral projections) were obtained but were not diagnostic. Rhinoscopy with a 2.7 mm rigid endoscope passed via the nostrils was performed revealing no abnormalities in the right nasal cavity, however, in the left nasal cavity at a depth of approximately 3 cm there was a mass estimated to be 1 cm3 in size, appearing to be based caudally. The surrounding nasal tissues were visibly inflamed. Nasal flush was performed to clear any discharge before biopsy to obtain material for histopathology. As the drainage lymph nodes were small no attempt was made to perform fine needle aspiration at this stage, invasive biopsy remaining an option if the nasal biopsy confirmed neoplasia. The dog was discharged with a course of meloxicam for analgesia (Metacam; Boehringer Ingelheim Vetmedi-ca GmbH, Germany; 0.1 mg/kg/day p.o.). Histopathology results andfurther treatment Histopathology results revealed an inflamed low-grade malignant nasal tumour, composed of two distinct subtypes predominant papillary and cystic adenocarcinoma with mucus secretion and formation of small cysts (Figure 1) and a smaller part of the transitional carcinoma, which is also referred to as respiratory epithelial carcinoma or nonkerati-nizing squamous cell carcinoma (Figure 2) (4). The adenocarcinomatous part was mostly composed of Figure 1: Papillary cystic adenocarcinoma. Part of the tumour shows a less well differentiated tall columnar epithelium with mild cellular and nuclear pleomorphism. A group of pleomorphic epithelial cells can be seen in the middle (arrow) and a mitotic figure in the left lower corner (arrow). There is abundant lymphocytic infiltrate and neu-trophils in the stroma. HE staining, x200 Figure 2: Transitional carcinoma. Cellular and nuclear pleomorphism is clearly evident. Abundant lymphocytic infiltration can be seen in the stroma. HE staining, x200 106 A. Nemec, Z. Pavlica, P. Juntes, D. A. Crossley well differentiated cuboidal or tall columnar epithelial cells with mild pleomorphism, only limited areas showing greater pleomorphism. The cells of a transitional carcinoma subtype revealed greater variability of cell shape and size, from smaller basaloid cells to larger tall columnar and spindle-shaped or polygonal cells with moderate amount of pale eosi-nophilic cytoplasm and hyperchromatic nuclei containing one or two, and rarely several, small nucleoli. Mitotic figures were rare in both parts. The dog was much better while on meloxicam and the owner was advised to proceed with staging. In view of the final diagnosis the owner was offered referral for a CT scan prior (3, 5) to possible radiation therapy. Discussion Periodontal disease is the most common chronic infectious disease in dogs affecting a majority of the mature population (6, 7), with small breeds being predisposed to it (8). Tumours of the nasal and paranasal sinuses are rare in most domestic species but are recognised most frequently in dogs. The prevalence, however, is only 0.3 to 2.4%, with medium to large dolichocephalic breeds being more often affected. The higher risk associated with a long nose may be related to the larger surface area of nasal epithelium and with the filtering capability, although a genetic basis in some breeds is also suspected (9). Incidence of nasal tumours also increases with age of the dog, the mean age being reported as 9 to 10 years (10, 11). Despite the low prevalence, however, Tasker (12) reports that neoplasia is the most common diagnosis in dogs with persistent nasal disease (one third of cases), where as periodontal disease is only recognized as the cause in 10% of cases. Ade-nocarcinoma is the most frequent malignant nasal tumour recognized in dogs (9, 10) with transitional carcinoma being the second most common, both being rare or not reported in other animal species (4). Acinic cell carcinoma or even neuroendocrine carcinoma can not been ruled out completely in this case as differentiation requires immunohistochemistry, which was not performed. As neuroendocrine carcinoma is an uncommon sinonasal tract neoplasm with aggressive clinical behaviour (13, 14) this differential diagnosis was not consistent with clinical and histomorphological findings in the presented case. The diagnostic approach to a patient with nasal discharge includes obtaining a complete history supported by thorough physical examination and routine blood tests to rule out systemic disease (1). If the results are normal as in the presented case, complete oral examination is the next step as well as nasal swabs for cytology and culture if fungal disease is suspected, before proceeding to imaging and rhinoscopy with biopsy sampling (1). However, blood test results may be normal in dogs with nasal neo-plasia as paraneoplastic disorders associated with nasal tumours are rare in dogs (10). As advanced periodontal disease with oronasal fistulae at the maxillary canine teeth was detected in this case, the chronic serous bilateral nasal discharge was suspected to be of dental origin, especially when the nasal discharge completely disappeared on the right side after the first dental treatment (extraction of the right maxillary canine tooth and closure of the fistula at that site), but remained on the untreated site. As the discharge from the left nostril also temporarily disappeared after extraction of the left maxillary canine tooth and antibiotic and anti-inflammatory treatment, dental disease must have had an influence on the nasal discharge, though the improvement may have been due to suppression of secondary bacterial infection in the nasal cavity with the use of antibiotics (11). After unilateral recurrence of the signs, inadequate healing of the oronasal fistula was considered the most likely differential diagnosis (15). Once this was ruled out further investigation was required to identify the cause. Nasal neoplasia most often presents initially with unilateral nasal discharge, epistaxis, epiphora and facial deformity occurring in more advanced cases (11). When there is only serous discharge and expiratory stertor, as our case, chronic rhinitis and nasopharyngeal dysfunction have also to be considered. It is impossible to say, what the primary disease was or if there is any link between the two diseases in the present case as periodontal disease is extremely common in older small breed dogs and adenocarcinoma, although not a common condition, is seen most often in older dogs. Both conditions have chronic courses, clinical signs persisting for months (1, 6, 10, 11). It is well established that chronic inflammation and/or infection with certain organisms (particularly toxin producing spirochetes) prediposes to carcinoma in certain sites but there have not been any reports suggesting a link between periodontitis and nasal carcinoma (16, 17). Oral lichen planus, a chronic inflammatory disease seen in man is reported to be clinically associated with Advanced periodontal disease in a Yorkshire terrier with concurrent nasal cavity malignancy 107 development to oral cancer, most likely due to oxidative and nitrative DNA damage caused by chronic inflammation (17-19). Therefore it is possible that the two diseases in the reported case could be related as it has been established that cancer can be promoted and/or exacerbated by inflammation and infections and vice versa (17, 20, 21). The cytokines produced by activated innate immune cells are reported to be important components in this linkage (20, 22-24), many of them are also elevated in periodontal disease (25). TNF-a can contribute to tumour initiation by stimulating production of genotoxic molecules, such as nitric oxide (NO) and reactive oxygen species (ROS), that can induce DNA damage and mutations (20). Stimulated polymorphonuclear leucocytes (PMN), increased in periodontal disease, can also produce reactive oxygen and nitrogen species (ROS, RNS) (17, 25). In periodontal disease there is also a marked production of NO in gingival tissues (26-28). Additionally, activation of toll-like receptors (TLR) on host macrophages or directly on tumour cells by bacterial lipopolysaccharides (LPS), which leads to the activation of NF-kB signaling pathway can enhance tumour development and to a lesser extent tumour regression as well (17, 20, 29). Bacteria can add to cancer development also by production of carcinogenic metabolites (16). After diagnosing the nasal mass, the dog was placed on meloxicam as the drug not only relieves pain and inflammation but also has the potential to slow the progression of some tumours this effect being suggested to be through the selective inhibition of cyclooxygenase-2 (COX-2), which is upregulated in epithelial nasal tumours (30, 31). Whilst side effects may occur with long term use of meloxicam (32), the risk is so low that it does not outweigh the potential benefits of its use. The owners of the dog have been recommended to consider more definitive treatment for the tumour, i.e. radiation therapy, as this currently provides the best results for nasal carcinomas (10). References 1. Cooke K. Sneezing and nasal discharge. In: Ettinger SJ, Feldman EC, eds. Textbook of veterinary internal medicine. 6th ed. StLouis, Missouri: Elsevier Saunders, 2005: 207 - 210. 2. MacEwen EG, Withrow SJ, Patnaik AK. Nasal tumors in the dog: retrospective evaluation of diagnosis, prognosis, and treatment. J Am Vet Med Assoc 170 (1), 1977: 45-8. 3. Lefebvre J, Kuehn NF, Wortinger A. Computed tomography as an aid in the diagnosis of chronic nasal disease in dogs. J Small Anim Pract 46 (6), 2005: 280-5. 4. Dungworth DL, Hauser B, Hahn FF, Wilson DW, Haenichen T, Harkema JR. Histological classification of tumours of the respiratory system of domestic animals. WHO International Histological Classification of tumours of domestic animals, Second Series, Volume VI. Washington: Armed Forces Institute of Pathology, 1999: 16-23. 5. Park RD, Beck ER, LeCouteur RA. Comparison of computed tomography and radiography for detecting changes induced by malignant nasal neoplasia in dogs. J Am Vet Med Assoc 201 (11), 1992: 1720-4. 6. DeBowes LJ. The effects of dental disease on systemic disease. Vet Clin North Am Small Anim Pract 28 (5), 1998: 1057-62. 7. DeBowes LJ, Mosier D, Logan E, Harvey CE, Lowry S, Richardson DC. Association of periodontal disease and histologic lesions in multiple organs from 45 dogs. J Vet Dent 13 (2), 1996: 57-60. 8. Harvey CE, Shofer FS, Laster L. Association of age and body weight with periodontal disease in North American dogs. J Vet Dent 11 (3), 1994: 94-105. 9. Hayes HM, Jr., Wilson GP, Fraumeni HF, Jr. Carcinoma of the nasal cavity and paranasal sinuses in dogs: descriptive epidemiology. Cornell Vet 72 (2), 1982: 168-79. 10. Fox LE, King RR. Cancers of the respiratory system. In: Morrison WB, eds. Cancer in dogs and cats. Medical and surgical management. 2nd ed. Jackson, Wyoming: Teton NewMedia, 2002: 497 - 512. 11. Ogilvie GK, LaRue SM. Canine and feline nasal and paranasal sinus tumors. Vet Clin North Am Small Anim Pract 22 (5), 1992: 1133-44. 12. Tasker S, Knottenbelt CM, Munro EA, Stonehewer J, Simpson JW, Mackin AJ. Aetiology and diagnosis of persistent nasal disease in the dog: a retrospective study of 42 cases. J Small Anim Pract 40 (10), 1999: 473-8. 13. Babin E, Rouleau V, Vedrine PO, et al. Small cell neuroendocrine carcinoma of the nasal cavity and paranasal sinuses. J Laryngol Otol 120 (4), 2006: 289-97. 14. Tarozzi M, Demarosi F, Lodi G, Sardella A, Carrassi A. Primary small cell carcinoma of the nasal cavity with an unusual oral manifestation. J Oral Pathol Med 36 (4), 2007: 252-4. 15. Holmstrom SE, Frost Fitch P, Eisner ER. Veterinary dental techniques for the small animal practitioner. 3rd ed. Philadelphia, Pennsylvania: Saunders, 2004. 16. Parsonnet J. Bacterial infection as a cause of cancer. Environ Health Perspect 103 Suppl 8, 1995: 263-8. 17. Kawanishi S, Hiraku Y. Oxidative and nitrative DNA damage as biomarker for carcinogenesis with special reference to inflammation. Antioxid Redox Signal 8 (5-6), 2006: 1047-58. 18. Chaiyarit P, Ma N, Hiraku Y, et al. Nitrative and oxidative DNA damage in oral lichen planus in relation to human oral carcinogenesis. Cancer Sci 96 (9), 2005: 553-9. 108 A. Nemec, Z. Pavlica, P. Juntes, D. A. Crossley 19. Kawanishi S, Hiraku Y, Pinlaor S, Ma N. Oxidative and nitrative DNA damage in animals and patients with inf lammatory diseases in relation to inf lammation-relat-ed carcinogenesis. Biol Chem 387 (4), 2006: 365-72. 20. Lin WW, Karin M. A cytokine-mediated link between innate immunity, inf lammation, and cancer. J Clin Invest 117 (5), 2007: 1175-83. 21. Shacter E, Weitzman SA. Chronic inflammation and cancer. Oncology (Williston Park) 16 (2), 2002: 217-26, 229; discussion 230-2. 22. Okamoto T, Sanda T, Asamitsu K. NF-kappa B signaling and carcinogenesis. Curr Pharm Des 13 (5), 2007: 447-62. 23. Rose-John S, Schooltink H. Cytokines are a therapeutic target for the prevention of inf lammation-induced cancers. Recent Results Cancer Res 174, 2007: 57-66. 24. Rhodus NL, Cheng B, Myers S, Miller L, Ho V, On-drey F. The feasibility of monitoring NF-kappaB associated cytokines: TNF-alpha, IL-1alpha, IL-6, and IL-8 in whole saliva for the malignant transformation of oral lichen planus. Mol Carcinog 44 (2), 2005: 77-82. 25. Sculley DV, Langley-Evans SC. Salivary antioxi-dants and periodontal disease status. Proc Nutr Soc 61 (1), 2002: 137-43. 26. Ugar-Cankal D, Ozmeric N. A multifaceted molecule, nitric oxide in oral and periodontal diseases. Clin Chim Acta 366 (1-2), 2006: 90-100. 27. Skaleric U, Gaspirc B, McCartney-Francis N, Masera A, Wahl SM. Proinflammatory and antimicrobial nitric oxide in gingival fluid of diabetic patients with periodon-tal disease. Infect Immun 74 (12), 2006: 7010-3. 28. Leitao RF, Ribeiro RA, Chaves HV, Rocha FA, Lima V, Brito GA. Nitric oxide synthase inhibition prevents alveolar bone resorption in experimental periodontitis in rats. J Periodontol 76 (6), 2005: 956-63. 29. Medzhitov R. Toll-like receptors and innate immunity. Nat Rev Immunol 1 (2), 2001: 135-45. 30. Kleiter M, Malarkey DE, Ruslander DE, Thrall DE. Expression of cyclooxygenase-2 in canine epithelial nasal tumors. Vet Radiol Ultrasound 45 (3), 2004: 255-60. 31. Knottenbelt C, Chambers G, Gault E, Argyle DJ. The in vitro effects of piroxicam and meloxicam on canine cell lines. J Small Anim Pract 47 (1), 2006: 14-20. 32. Luna SP, Basilio AC, Steagall PV, et al. Evaluation of adverse effects of long-term oral administration of carpro-fen, etodolac, flunixin meglumine, ketoprofen, and meloxi-cam in dogs. Am J Vet Res 68 (3), 2007: 258-64. NAPREDOVALA PARODONTALNA BOLEZEN IN MALIGNI TUMOR NOSNE VOTLINE PRI JORKŠIRSKEM TERIERJU A. Nemec, Z. Pavlica, P. Juntes, D. A. Crossley Povzetek: Enajst let star, 2,6 kilograma težek jorkširski terier, ki živi pretežno v stanovanju, je bil napoten na kardiološki pregled zaradi 6 mesecev trajajočih težav z dihanjem. Pri pregledu kardiopulmonarne težave niso bile ugotovljene, pes pa je imel težave zaradi kroničnega seroznega nosnega izcedka ter sprememb v ustni votlini. Pregled ustne votline v splošni anesteziji je potrdil sum napredovale parodontalne bolezni z oronazalnima fistulama ob zgornjem desnem in levem grabilcu. Serozni nosni izcedek se je ponovil 1 mesec po sanaciji ustne votline, tokrat le iz leve nosnice. Ker oronazalne fistule ni bilo, je bila izvedena rinoskopija z odvzemom tkivnih bioptov mase v levi nosnici. Patohistološko je bil v levi nosni votlini dokazan maligni tumor, papilarno-cističen adenokarcinom, kombiniran s prehodnim karcinomom. Ključne besede: parodontalne bolezni; rinitis; nos, novotvorbe - patologija; adenokarcinom; psi Slov Vet Res 2007; 44 (4): 109-14 UDC 636.7.09:616.68-006-089:616-073+616-076 Case Report GENERALIZED SYMMETRIC ALOPECIA AND HYPEROESTRO-GENISM ASSOCIATED WITH CONCURRENT LYMPHOMA, SERTOLI CELL TUMOUR AND SEMINOMA IN A SAMOYED Tanja Plavec*, Nataša Tozon, Tina Kotnik Clinic for Small Animal Medicine and Surgery, Veterinary Faculty, Gerbičeva 60, 1000, Ljubljana, Slovenia Corresponding author, E-mail: tanja.plavec@vf.uni-lj.si Summary: A 10-year-old Samoyed with unilateral right cryptorchidism was referred to the Clinic for Small Animal Medicine and Surgery, Veterinary Faculty of Ljubljana University, due to a symmetrical, noninflammatory, mildly pruritic alopecia of 6-month duration. It had a history of diarrhoea, which responded to cyclosporine treatment. Lymphoma and testicular neoplasia were suspected based on ultrasonography and fine needle aspiration cytology. The dog was castrated and splenic and gastric lymph node biopsies were obtained. Histopathology revealed three different tumours: Sertoli cell tumour and seminoma were present in the right inguinal testicle, and B cell lymphoma was present in the spleen and lymph node. After two months when the peripheral lymph nodes were considerably enlarged and the owners declined chemotherapy, they were advised to start corticosteroid treatment. Three months after the castration, the hair coat looked normal. Four months after the castration, the dog was euthanized at the owner's request by the referring veterinarian due to a lymphoma related disease. Key words: unilateral cryptorchidism; testicular neoplasms - surgery - chemotherapy; ultrasonography; cytodiagnosis; biopsy, fine-needle; dogs Introduction Noninflammatory alopecias are quite common in dogs and can be congenital, hereditary or acquired. The latter can result from endocrinopathies (e.g. hyper-adrenocorticism, hypothyroidism, sexual imbalance), telogen or anagen effluvium, metabolic imbalance (disruptions in protein or fatty acid metabolism) and idi-opathic disturbances of the hair growth cycle (1). Testicular tumours comprise 4-7% of all tumours in male dogs (2). They arise either from Sertoli cells (i.e. Sertoli cell tumours), germinal epithelium (i.e. seminoma) or interstitial cells (i.e. Leydig cell tumours) (3), all of which occur with equal prevalence (2, 4). Other primary tumours may also arise, but are extremely rare. Dogs with non-descended testicles have an increased risk of up to 13.6 fold to develop a testicular tumour compared to dogs with descended testicles (5). Since right-sided cryptorchidism is Received: 10 July 2007 Accepted for publication: 17 September 2007 more common, the right testis is more often affected (4). Multiple tumours within one testis occur in up to 46% of the cases and are also described in mac-roscopically normal testeses (6). Hyperestrogenism occurs in 25-50% of dogs with Sertoli cell tumour and less frequently with seminomas, and may lead to signs of feminization (7). It can lead to bone marrow hypoplasia with consecutive thrombocytopenia, non-regenerative anaemia and granulocytopenia. Serum testosterone and progesterone concentrations may be increased, but usually testosterone concentration is low to undetectable (5). As these changes may be subtle and are not specific and sensitive changes, castration with a histopathological examination is still the diagnostic and therapeutic approach of choice (8). An alternative and perhaps more sensitive marker for canine Sertoli cell tumour is increased serum inhibin concentration (9). Malignant lymphoma is the most common canine haemato-lymphatic tumour, most commonly classified based on anatomic location, clinical stage or immunophenotype. The multicentric form with 110 T. Plavec, N. Tozon, T. Kotnik peripheral lymphadenopathy is the most common form, followed by the alimentary, mediastinal and extranodal (kidneys, skin, and brain) forms (10-12). The aetiology of canine lymphoma is unknown, but hypotheses include retroviral infection, exposure to herbicides and magnetic field, chromosomal abnormalities and immune dysfunction (13). Case report A 10-year-old, intact male Samoyed with unilateral right cryptorchidism was presented at the Clinic for Small Animal Medicine and Surgery, Veterinary Faculty of Ljubljana University for an investigation of a symmetric alopecia of 6-month duration. It was currently vaccinated and dewormed and has lately gained weight although water and food intake seemed unchanged. The dog had a history of large bowel diarrhoea eight months before the presentation at our clinic, which was treated with metronidazole and spiramycin for 1 month without obvious improvement. This was followed by cyclosporine treatment for 40 days, which led to complete recovery, however, dermatologic abnormalities were observed. Initially, alopecia was present only in the neck region, and later symmetrically spread to the axillae, abdomen as well as the inguinal and tail areas. Mild pruritus was observed during the disease course. An abdominal ultrasound performed by the referring veterinarian did not reveal abnormalities. Serum hormone concentrations were determined and included free T4 (1.1 ng/dl, reference range 0.6 - 3.7), cTSH (0.13 ng/ ml reference range <0.40). testosterone (0.0667 ng/ ml, reference range 0.3 - 5.8)1 and 17-p-oestradiol (87.9 pg/ml, reference range <15 (9)). Physical examination revealed, in addition to the alopecia (Figure 1 and 2) and cryptorchidism, a bilateral purulent ocular discharge. Skin scrapings for Sarcoptes and Demodex were negative. Cytology of the skin was positive for Malassezia. Faecal examination was positive for Giardia. CBC (Bayer Tech-nicon H*1, Bayer-Technicon, Germany) and serum biochemistry analysis (biochemical analyzer Tech-nicon RA-XT, Bayer Technicon, Germany) including glucose, urea, creatinine, calcium, phosphorous, alkaline phosphatase, alanine-aminotransferase and cholesterol) were unremarkable, with exclusion of a mild normocytic normochromic anaemia (haemato-crit 0.35 l/l, reference range 0.37-0.55 (14)). Figure 1: Skin at the beginning of the therapy Figure 2: Skin at the beginning of the therapy 1 References values of the Vet Med Lab laboratory in Trieste, Italy. Figure 3: Hair re-growth after 3 weeks of the hypoallergen-ic diet and essential fatty acid supplementation Generalized symmetric alopecia and hyperoestrogenism associated with concurrent lymphoma, Sertoli cell tumour 111 Figure 4: Lymphoblasts in the spleen (x1000) Figure 5: Dog three months after surgery Figure 6: Dog three months after surgery Treatment was initiated using anthelmintics praziquantel 5 mg/kg and fenbendazole 50 mg/kg (Zantel, CAM) PO q24h for 5 days, ketoconazole 14 mg/ kg PO q24h (Oronazol, Krka) and topical ophthalmic gentamicin (Garamycin, Krka) ointment bilaterally q12h. After one month, there was no improvement and total serum T4 and TSH analyses were repeated (30.6 nmol/l and 0.47 ng/ml, respectively). A hypoal-lergenic diet (Eucanuba FP dermatosis and homemade diet (horse meat, rabbit meat, fish) and an oral omega 3 and omega-6 fatty acids preparation (Der-manorm, Vetoquinol, 2 capsules/dog PO q24h) were prescribed for three weeks, when an improvement in general health and growth of hair was seen (Figure 3). The same therapy was continued for 2 additional months. At that time, the dog was rechecked due to deterioration in the condition of the skin. It had pruritus, alopecia, erythema, ventral hyper- and hypo-pigmentation and dorsal hypotrichia, seborrhoea and crusts. An abdominal ultrasound revealed nonhomo-geneous splenomegally and nonhomogenous masses: two cranio-dorsal to right and two cranio-dorsal to the left kidney. These masses were interpreted as lymph nodes by the radiologist. The testicle present within the scrotum had a hypoechoic discrete nodule of 8 mm diameter. A nonhomogeneous 3*4 cm mass in the right inguinal area was observed. Other abdominal organs were ultrasonografically normal. A fine needle aspiration of the inguinal mass (probably testicle) and the spleen was performed. The first was non-diagnostic while the latter revealed a population of large lymphoid cells with large nuclei, scant deeply blue cytoplasm, some of which had a pronounced perinuclear reactive zone and 1-3 prominent nucleoli of different size and shape (Figure 4). According to these findings we planned to castrate the dog and take samples of the spleen, enlarged lymph nodes and skin. A preoperative CBC showed a mild normochromic normocytic nonregenerative anaemia (RBC 5.02 * l012/l, reference range 5.5-8.5 * 1012/l; haemoglobin 115 g/l, reference range 120-180 g/l; haemat-ocrit 0.32 l/l, reference range 0.37-0.55 l/l) and mild thrombocytopenia (platelets 141 * 109/l, reference range 200-500 * 109/l)(14). Serum biochemistry was unremarkable. Under general anaesthesia, the dog was castrated and the right inguinal mass was excised. This mass was 5 cm in diameter and had an appearance of a testicle. Then, a midline exploratory laparotomy was performed. The spleen as well as the gastric lymph node were enlarged and were removed. The mesenteric lymph nodes were also 112 T. Plavec, N. Tozon, T. Kotnik mildly enlarged (5 cm diameter). No other abnormalities were observed. The biopsy of the skin was also performed. Histopathology of the right inguinal mass revealed the presence of seminoma and Sertoli cell tumour in the right testis. The left testis was not examined. Lymphoma was present in the spleen and gastric lymph node. Subchronic dermatitis with a mixed cell (predominantly eosinophils) population was present in the skin biopsies. Immunohisto-chemistry examination of splenic and gastric lymph node biopsies with CD3 and CD79 antibodies (DAKO REALTM, EnVision Detection System) confirmed a B cell lymphoma. Postoperatively a bloody-purulent urine was noticed. Dipstick and urine sediment analyses confirmed a diagnosis of purulent cystitis. Culture and sensitivity were not performed. Amoxicillin with cla-vulanic acid (Synulox, Pfizer) 20 mg/kg PO q12h for 14 days was prescribed. The dog was rechecked one month later, seemed healthy, gained two kilograms of body weight and its hair-coat showed signs of improvement. The palpable peripheral lymph nodes were unchanged and CBC was unremarkable (mild anaemia, haematocrit 0.36 l/l). A month later, the dog had mild mandibular and praescapular lymphadenopathy. CBC was normal, with the exception of a mild anaemia (haema-tocrit 0.34 l/l). The owners were offered a multidrug chemotherapy, but declined and were thus advised to start corticosteroid treatment - methylprednisolo-ne (Medrol, Pfizer; 2 mg/kg/day PO until relaps). An additional month later, three months after the operation, Lord's hair looked normal again (Figure 5, 6). A month later, after two months of methylpred-nisolone treatment, the dog deteriorated, was depressed and anorexic and was euthanized at the owner's request by the referring veterinarian. Discussion This is an unusual case with 3 different neopla-sias occurring concurrently in a single dog. Multiple tumours occurring in the brains (15, 16) or in the mammary tissue (17) were reported, but authors are not aware of any report about concurrent multiple tumour occurrence in different tissues. Preoperatively seen mild nonregenerative anaemia with the haematocrit of 0.32 l/l and thrombo-cytopaenia could have developed secondarily to lymphoma (18,19), but haematocrit of 0.35 l/l was also seen 3 months earlier, when lymphoma was not diagnosed yet. Such a long course of the disease is unusual for lymphoma (18). In addition to symmetrical truncal alopecia other male patients, but not the present case, may also present other signs of feminization (8). The subsequent improvement of the skin after surgery in contrast to the partial response to previous therapy supports the assumption that these changes were secondary to hyperoestrogenism due to Sertoli cell tumour. Plasma oestradiol concentrations in male dogs should be less than 15 pg/ml. In dogs with Sertoli cell tumours, eos-tradiol concentrations are usually 10 to 150 pg/ml, as was in the present case (87,9 pg/ml) (9). Sertoli cell tumours metastasize in 2-14% of cases; seminomas do so even less frequently. Most commonly they metastasize to the sublumbar (3), inguinal and iliac lymph nodes, lungs, liver, spleen, kidneys, pancreas (9), subcutis, brain and eyes (20). The above mentioned lymph nodes were not enlarged in our case, as observed upon the exploratory laparotomy. However, gastric and mesenteric lymphadenopathy was observed. While the gastric lymph node presented lymphoma and had no evidence of a testicular cancer metastasis, the mesenteric lymph nodes were not examined his-tologically. Although we cannot rule out that they were enlarged due to metastasis of the testicular tumours, it is more likely that lymphoma was the cause. A postoperative serum oestrogen could have aided in determination of Sertoli cell tumour metastasis. Retrospectively, considering the chronic diarrhoea, unresponsive to antibiotics, but responsive to cyclosporine, the mesenteric and gastric lymphad-enopathy, and diagnosis of lymphoma in the latter as well as in the spleen, we believe this was a case of alimentary lymphoma. It is presumed that they are most likely B-cell in origin (18), but newer studies are conflicting (21). However, it could be also a stage IVa of the multicentric form (18), which is perhaps more likely regarding subsequent development of generalised lymphadenopathy. The chronic diarrhoea, mentioned in the history, could develop due to other causes such as lym-phocytic-plasmacitic enteritis (LPE) or giardiasis, as the dog was positive for the latter. Giardiasis seems to be a less likely cause for the diarrhoea as the dog failed to respond to a prolonged metronidazole and spiramycin treatment, and did respond to cy-closporine (22, 23). It has been suggested, that LPE may be a prelymphomatous change in the gastrointestinal tract (18). Cyclosporine treatment has been reported to induce lymphoma or other tumours oc- Generalized symmetric alopecia and hyperoestrogenism associated with concurrent lymphoma, sertoli cell tumour 113 casionally (18, 24, 25). The drug hinders lymphocyte T activity and recognition of neoplastically changed cells (26), which, presumably, can consequently induce additional concurrent neoplastic disease. Dogs with lymphoma are known to be less immu-nocompetent because of the incompetence of neo-plastic lymphocytes/lymphoblasts (11, 18) or fewer total lymphocytes, especially T cells (27). This could have led to the observed postoperative cystitis. Lymphoma is usually treated with combination chemotherapy protocol (28-30), rather than corti-costeroids a single agent, however, the latter was attempted due to the owner's decline of the former. Corticosteroid effects are usually short-lived in canine lymphoma, and relapses are extremely common (18), and thus it is not surprising that the present dog was represented shortly later with signs of systemic disease, most probably a relapse of the lymphoma. Conclusion We presented a case of multiple neoplasia in a dog including B cell lymphoma, Sertoli cell tumour and seminoma. The dog had a chronic dermatopa-thy, probably due to hyperoestrogensim secondary to the Sertoli cell tumour. The latter is supported by recovery of the skin following castration. The lymphoma was probably alimentary, although a multi-centric lymphoma cannot be ruled out. The cause of lymphoma is unknown, but it may have been associated with a previous cyclosporine therapy. Acknowledgement We would like to thank Prof. Dr. Polona Juntes for her contribution to the pathohistologic diagnosis and Assist. Prof. Dr. Aleksandra Domanjko Petric for the ultrasonographic examination. References 1. Heripret D. Alopecia. In: Ettinger SJ, Feldman EC, eds. Textbook of veterinary internal medicine. 6th ed. St. Louis: Elsevier Saunders, 2005: 34-7. 2. Spugnini EP, Bartolazzi A, Ruslander D. Seminoma with cutaneous metastases in a dog. J Am Anim Hosp Assoc 2000; 36 (3): 253-6. 3. Cooley DM, Waters DJ. Tumors of the male reproductive system. In: Withrow SJ, MacEwen EG, eds. Small animal clinical oncology. 3rd ed. Philadelphia: Saunders, 2001: 478-89. 4. Kim O, Kim KS. Seminoma with hyperestrogenemia in a Yorkshire terrier. J Vet Med Sci 2005; 67 (1): 121-3. 5. Morrison WB. Cancer of the reproductive tract. In: Morrison WB, ed. Cancer in dogs and cats: medical and surgical management. 2nd ed. Jackson: Teton NewMedia, 2002: 555-64. 6. Peters MAJ, de Rooij DG, Teerds KJ, van der Gaag I, van Sluijs FJ. Spermatogenesis and testicular tumours in ageing dogs. J Reprod Fertil 2000; 120 (2): 443-52. 7. Chun R, Garrett. Urogenital and mammary gland tumors. In: Ettinger SJ, Feldman EC, eds. Textbook of veterinary internal medicine. 6th ed, St. Louis: Elsevier Saunders, 2005: 784-9. 8. Scott DW, Miller WH, Griffin CE. Small animal dermatology. 6th ed. Philadelphia: W.B. Saunders, 2001: 780-885. 9. Feldman EC, Nelson RW. Canine and feline endocrinology and reproduction. 3rd ed. St. Louis: Saunders, 2003: 961-76. 10. Tozon N, Samardzija P, Prijic S, Fazarinc G. Canine lymphoma: cytologic study and response to therapy. Slov Vet Res 2006; 43 (3): 127-33. 11. Vonderhaar MA, Morrison WB. Lymphosarcoma. In: Morrison WB, ed. Cancer in dogs and cats: medical and surgical management. 2nd ed. Jackson: Teton NewMedia, 2002: 641-70. 12. Lowe AD. Alimentary lymphosarcoma in a 4-year-old Labrador retriever. Can Vet J 2004; 45 (7): 610-2. 13. Fan TM. Lymphoma updates. Vet Clin North Am Small Anim Pract 2003; 33 (3): 455-71. 14. Bush BM. Interpretation of laboratory results for small animal clinicians. Oxford: Blackwell Science, 1991: 35-131. 15. Stacy BA, Stevenson TL, Lipsitz D, Higgins RJ. Simultaneously occurring oligodendroglioma and meningi-oma in a dog. J Vet Int Med 2003; 17 (3): 357-9. 16. Alves A, Prada J, Almeida JM, et al. Primary and secondary tumours occurring simultaneously in the brain of a dog. J Small Anim Pract 2006; 47 (10): 607-10. 17. Ferguson HR. Canine mammary gland tumors. Vet Clin North Am Small Anim Pract. 1985; 15 (3): 501-11. 18. Vail DM, MacEwen EG, Young KM. Canine lymphoma and lymphoid leukemias. In: Withrow SJ, MacEwen EG, eds. Small animal clinical oncology. 3rd ed. Philadelphia: Saunders, 2001: 558-90. 19. Gaschen FP, Teske E. Paraneoplastic syndrome. In: Ettinger SJ, Feldman EC, eds. Textbook of veterinary internal medicine. 6th ed, St. Louis: Elsevier Saunders, 2005: 789-95. 20. Dhaliwal RS, Kitchell BE, Knight BL, Schmidt BR. Treatment of aggressive testicular tumors in four dogs. J Am Anim Hosp Assoc 1999; 35 (4): 311-8. 21. Coyle KA, Steinberg H. Characterization of lymphocytes in canine gastrointestinal lymphoma. Vet Pathol 2004; 41 (2): 141-6. 22. Leib MS, Dalton MN, King SE, Zajac AM. Endoscop- 114 T. Plavec, N. Tozon, T. Kotnik ic aspiration of intestinal contents in dogs and cats: 394 cases. J Vet Int Med 1999; 13 (3): 191-3. 23. Lappin MR. Giardia infections. In: World congress 2006 WSAVA/FECAVA/CSAVA. Proceedings of the 31st World Small Animal Veterinary Association. Prague: WSA-VA, 2006: 50-1. 24. Blackwood L, German AJ, Stell AJ, O'Neill T. Mul-ticentric lymphoma in a dog after cyclosporine therapy. J Small Anim Pract 2004; 45 (5): 259-62. 25. Callan MB, Preziosi D, Mauldin E. Multiple papil-lomavirus-associated epidermal hamartomas and squa-mous cell carcinomas in situ in a dog following chronic treatment with prednisone and cyclosporine. Vet Derma-tol 2005; 16 (5): 338-45. 26. Knapp DW: Immunotherapy and biologic response modifiers. In: Morrison WB, ed. Cancer in dogs and cats: medical and surgical management. 2nd ed. Jackson: Teton NewMedia, 2002: 425-39. 27. Walter CU, Biller BJ, Lana SE, Bachand AM, Dow SW. Effects of chemotherapy on immune responses in dogs with cancer. J Vet Int Med 2006; 20 (2): 342-7. 28. Chun R, Garrett LD, Vail DM. Evaluation of a highdose chemotherapy protocol with no maintenance therapy for dogs with lymphoma. J Vet Intern Med 2000; 14 (2): 120-4. 29. Garrett LD, Thamm DH, Chun R, Dudley R, Vail DM. Evaluation of a 6-month chemotherapy protocol with no maintenance therapy for dogs with lymphoma. J Vet Intern Med 2002; 16 (6): 704-9. 30. Simon D, Nolte I, Eberle N, Abbrederis N, Killich M, Hirschberger J. Treatment of dogs with lymphoma using a 12-week maintenance-free combination chemotherapy protocol. J Vet Intern Med 2006; 20 (4): 948-54. GENERALIZIRANA SIMETRIČNA ALOPECIJA IN HIPERESTROGENIZEM PRI SAMOJEDU Z LIMFOMOM, SEMINOMOM IN TUMORJEM SERTOLIJEVIH CELIC T. Plavec, N. Tozon, T. Kotnik Povzetek: 10 let star samojed, desnostranski kriptorhid, je bil napoten na Kliniko za kirurgijo in male živali Veterinarske fakultete zaradi šest mesecev trajajoče nevnetne alopecije. V anamnezi živali so lastniki navajali drisko, ki je bila uspešno zdravljena s ciklosporinom. Na podlagi ultrazvočne preiskave in citološke preiskave tankoigelnega punktata smo postavili sum na limfom in neoplazijo mod. Psa smo kastrirali ter odvzeli biopsat vranice in bezgavke. Patohistološka preiskava je potrdila tumor sertolijevih celic in seminom desnega moda ter B celični limfom vranice in bezgavk. Dva meseca po operativnem posegu smo opazili povečane periferne bezgavke, vendar se lastniki niso odločili za citostatično zdravljenje. Svetovali smo jim začetek zdravljenja s kortikosteroidi. Tri mesece po kirurškem posegu je bila odlakanost psa spet normalna, evtanaziran je bil štiri mesece po kastraciji zaradi obolenja, povezanega z limfomom. Ključne besede: enostranski kriptorhidizem; testis, novotvorbe - kirurgija - kemoterapija; ultrazvok; citodiagnostika; biopsija, tankoigelna; psi Slov Vet Res 2007; 44 (4): 115-22 UDC 636.7.09:616.579.842:616.98 Case Report NOSOCOMIAL KLEBSIELLA OXYTOCA INFECTION IN TWO DOGS Alenka Seliskar1*, Irena Zdovc2, Bojan Zorko1 1 Clinic for Small Animal Medicine and Surgery, 2Institute for Microbiology and Parasitology, University of Ljubljana, Veterinary Faculty, Gerbiceva 60, 1000 Ljubljana, Slovenia Corresponding author, E-mail: alenka.seliskar@vf.uni-lj.si Summary: Despite a large amount of published data on nosocomially-acquired intravenous catheter-related Klebsiella oxytoca infection in human medicine there is a lack of information in veterinary medicine. Intravenous catheter-related Klebsiella oxytoca infection was strongly suspected in two dogs that underwent dental procedures under general anesthesia. Dog 1 was presented with severe osteomyelitis of the shoulder joint from the same leg that was used for intravenous catheter placement during implantation of bilateral direct acrylic inclined plane approximately 3 weeks ago. According to the bacteriology identification that revealed Klebsiella oxytoca infection and susceptibility testing of the synovial fluid from the shoulder joint the dog was treated with amoxicillin+clavulanic acid for one month. In spite of clinical improvement the radiographic examination revealed severe osteoarthritis of the affected joint at the end of the course. Dog 2 died of septic shock associated with disseminated intravascular coagulation 7 hours after the extraction of left mandibular fourth premolar tooth. Klebsiella oxytoca was isolated from several abdominal organs post-mortem. Bacteriology examination of hospital environment and equipment was carried out because of possible common-source epidemic and among other microorganisms, Klebsiella oxytoca was isolated. Additional infection control measures were instituted and so far, eigteen-months after that no nosocomial infection was confirmed. Key words: cross infection - microbiology; infection control; Klebsiella infections - etiology - microbiology; postoperative complications; dogs Introduction Nosocomial infections are infections that may be derived from endogenous flora of the patients or from exogenous microorganisms acquired by patients during their stay in the hospital (1, 2). In the latter case, the infectious agent is often resistant to multiple antibiotics and may be transmitted to or between several patients, thus leading to clusters or outbreaks of nosocomial infections due to the same strain (3). A limited number of small outbreaks is documented in companion animals due to Klebsiella sp. (4), Serratia marcescens (5), Enterobacter cloace (6), Escherichia coli (7, 8), Enterococcus faecium (2), Clostridium difficile (9, 10), Clostridium perfringens (11), Acinetobacter baumanni (2, 12) and Pseudomonas aeruginosa (13). In human medicine, Received: 6 August 2007 Accepted for publication: 29 October 2007 there are several documented outbreaks of catheter or contaminated disinfectant-associated infections with Klebsiella oxytoca (14, 15, 16, 17, 18, 19). The data on catheter associated Klebsiella oxytoca infection in veterinary medicine are sparse (4, 20). Bacteria from genus Klebsiella are non-motile, rod-shaped, Gram-negative aerobic bacteria that possess a prominent polysaccharide capsule. Some of these bacteria produce an extracellular toxic complex that has been shown to be lethal for and produce extensive lung pathology in mice. It is composed of capsular polysaccharide, lipopolysaccha-ride, and protein; when introduced to experimental animals in sublethal doses, the animals built up immunization due to antibody production (21). Kleb-siella oxytoca and Klebsiella pneumoniae are both opportunistic pathogens found in the environment and in mammalian mucosal surfaces; they are usually passed by the hands of hospital personnel. Kleb- 116 A. Seliskar, I. Zdovc, B. Zorko siella oxytoca and Klebsiella pneumoniae are common isolates in clinical microbiology and important producers of extended spectre beta-lactamases (ESBL). Enterobacteriaceae with beta-lactam resistance due to the production of ESBL were discovered in the eighties and since that time became epidemic and endemic in hospitals worldwide (22). The purpose of this report is to describe nosocomial Klebsiella oxytoca infection in two dogs that underwent dental procedures at Clinic for Small Animal Medicine and Surgery, Veterinary Faculty, University of Ljubljana (CSAMS-VFLJ). Case 1 Dog 1, a 7-months-old intact male Airedale terrier, considered healthy after physical examination, weighing 19.5 kg was presented to CSAMS-VFLJ for orthodontic movement of right mandibular canine tooth. Bilateral direct acrylic inclined plane was implanted under general anaesthesia. The dog was premedicated with medetomidine (Domitor, Pfizer, Karlsruhe, Germany) 0.018 mg/kg i/m. Intravenous catheter was placed into cephalic vein 15 minutes later after standard aseptic procedure including clipping the hair with the clipper and disinfection of the skin with a mixture of 2-propanol and ben-zalkonium chloride (Cutasept, Beiersdorf GsmbH, Wien, Austria). The dog was induced to general anaesthesia with propofol (Propofol 1%, Fresenius Kabi, Graz, Austria) 1.5 mg/kg i/v, endotracheally intubated and maintained with isoflurane (Forane, Abbott, Queenborough, UK) in 100% oxygen for 20 minutes. Atipamezole (Antisedan, Pfizer, Karlsruhe, Germany) 0.038 mg/kg i/m was administered at the end of the procedure to antagonize the effects of me-detomidine. During general anaesthesia, Lactated Ringer's solution (B Braun, Melsungen, Germany) was administered at 10 ml/kg/h. One hour and half after atipamezole administration, metoclopramide (Reglan, Alkaloid, Skopje, Macedonia) 0.4 mg/kg s/c was given to treat postoperative nausea. The dog was discharged to a home care few hours later. The dog was brought to local veterinary practitioner because of lameness of right front leg that was used for intravenous catheter placement the next day. Intense pain was observed at palpation of right shoulder joint and the dog was given carprofen (Rimadyl, Pfizer, Animal Health S.A., Dundee, UK) 4 mg/kg s/c. The lameness resolved the day after that, but the mild pain was still present at palpation of affected joint. Radiographic examination showed no abnormalities and the dog was prescribed carpro-fen (2 mg/kg p/o q12h). Five days later, the dog returned to veterinary practitioner with extensive swelling of right front leg. The dog was prescribed amoxicillin+clavulanic acid (Synulox, Pfizer Italiana, Latina, Italy) 20 mg/kg p/o q12h and carprofen as before. The dog's condition improved and therapy was terminated 10 days later. The day after, the dog was presented again with intense pain of right shoulder joint, painful hind part of the body and stiffed gait. The dog was reluctant to walk. According to veterinary practitioner, radiographic examinations of right shoulder joint and hip joints were normal as well as neurological examination. Routine haematology profile was within reference range. The dog was tested for dirofilari-osis, boreliosis and ehrlichiosis with commercially available ELISA kit (SNAP 3Dx, Canine Heartworm Antigen/Borrelia Burgdorferi/Ehrlichia Canis Ani-body Test Kit, IDEXX Laboratories, Westbrook, Maine, USA), and the results were negative. Despite negative results, the dog was prescribed doxycycline (Clinofug D, Dr. August WOLFF GmbH&Co, Bielfeld, Germany) 10 mg/kg p/o q24h and carprofen 2 mg/ kg p/o q12h. The dogs' condition worsened in the course of therapy, and a week later the dog was referred to CSAMS-VFLJ. The dog was presented to CSAMS-VFLJ with intense pain of the whole right front leg. Radiographic examination revealed osteomyelitis of the right humeral head. Definitive pattern of lysis and solid pattern of periosteal new bone were seen on the humeral head (Figure 1). Synovial fluid from right shoulder joint was taken and sent to bacteriology identification and susceptibility testing to commonly used antimicrobial agents. Bacteriology examinations were performed in Columbia agar supplemented with 5% of ovine blood. Klebsiella microorganisms were identified on the basis of colony morphology, microscopic Gram stain characteristics, indole and oxidase activity. The final confirmation was done with API-20E® enteric identification system (bioMerieux, Marcy l'Etoile, France). The antibiotic susceptibility of isolated strains was determined by disc-diffusion method according to the NCCLS (National Committee on Clinical Laboratory Standards) guidelines (Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals; Approved Standard - Second Edition, M31-A2, Vol. 22 No.6). Nosocomial Klebsiella oxytoca infection in two dogs 117 The following antimicrobials were assayed: azithromycin, amikacin, amoxicillin, amoxicillin/cla-vulanic acid, cephalexin, cephalotin, ciprofloxacin, enrofloxacin, ceftriaxone, gentamicin, neomycin, piperacilin, metronidazole, trimethoprim/sulfame-thoxazole and oxytetracycline. Initial beta-lactamase activity was detected by the hydrolysis of nitrocefin (cefinase). Screening tests for extended-spectrum-p-lactamase activity were evaluated regarding their susceptibility to cefpodoxime, ceftazidime, cefotaxime and cefuroxime (BBL-Difco). Confirmation tests for ESBL were done by E-test (E-test, AB- Biodisk, Dalvagen, Solna, Sweden) for cefotaxime/ cefotaxime+clavulanic acid (CT/CTL) and ceftazidime/ ceftazidime+clavulanic acid (TZ/TZL). Bacteriology identification revealed infection by Klebsiella oxytoca, p-lactamase positive by nitroce-fin testing, but ESBL negative (Table 1). According to the susceptibility testing, the dog was prescribed amoxicillin+clavulanic acid 20 mg/kg p/o q12h for one month and carprofen 2 mg/kg p/o q12h, the dose of latter to be gradually decreased by the owner according to the intensity of pain. Dog 1 Dog 2 osteomyelitis deceased Klebsiella oxytoca (synovial fluid) Klebsiella oxytoca (liver, kidney, spleen, peritoneal fluid, small intestine) Escherichia coli (small intestine) Susceptibility testing K. oxytoca K. oxytoca E. coli azithromycin R R R amikacin S S S amoxicillin R R I amoxicillin+clav.acid S S S cephalexin S I I cephalotin S S S ciprofloxacin S S S enrofloxacin S S S ceftriaxone S S gentamycin S S S neomycin S S S piperacillin S S metronidazole R R trimethoprim/sulpha S S S oxytetracycline S S cefpodoxime S S S ceftazidime S S S cefotaxime S S S cefuroxime S S S p-lactamase activity by + + + nitrocefin testing E-test, ESBL CT/CTL - - - E-test, ESBL TZ/TZL - - - Table 1: Antimicrobial susceptibility profile from microorganisms isolated from dog 1 and dog 2 S, sensitive; I, intermediate sensitive; R, resistant; E-test ESBL CT/CTL, extended specter p-lactamase, cefotaxime/ cefotaxime+clavulanic acid; E-test ESBL TZ/TZL, extended specter p-lactamase, ceftazidime/ ceftazidime+clavulanic acid; +, positive; -, negative 118 A. Seliskar, I. Zdovc, B. Zorko TWo weeks after the commencement of the therapy, the dog was much less in pain and able to use right front leg, although it was still lame. The dog was brought for a control check 6 weeks after the end of amoxicillin+clavulanic acid therapy. No lameness or pain was observed at the clinical examination and according to the owners it was doing well. Figure 1: Lateral radiograph of the right humeral head 3 weeks after the intravenous catheter-related infection with Klebsiella oxytoca Osteomyelitis with definitive pattern of lysis and solid pattern of periostal new bone is seen. Figure 2: Lateral radiograph of the right humeral head 6 weeks after the end of amoxicillin+clavulanic acid therapy (20 m/kg p/o q 12h for one month) shows resolution of the lytic process. The shape of the humeral head is irregular and severe osteoarthritis of the shoulder joint is visible. Radiographic examination of the right shoulder joint showed resolution of the lytic process. The shape of the humeral head was irregular and severe osteoarthritis of the shoulder joint was visible (Figure 2). Case 2 Dog 2, a 2 years-old intact working male German shepherd weighing 30.6 kg, considered healthy after physical examination including pulse and respiratory rate and heart and lung auscultation, was brought to CSAMS-VFLJ for extraction of left mandibular fourth premolar tooth three weeks after the dog 1. Body temperature was not measured because of the dog's un-cooperativeness. The extraction was performed under general anaesthesia with isoflurane in 100 % oxygen after premedication with medetomidine 0.025 mg/kg i/m and butorphanol (Torbugesic, Fort Dodge Animal Health, Iowa) 0.1 mg/kg i/m and induction to anesthesia with propofol at a dose of 2 mg/kg i/v. Intravenous catheter placement procedure was the same as in the dog 1. The dog was given perioperatively carprofen 4 mg/kg i/v and Lactated Ringer's solution at 10 ml/ kg/h. At the end of the procedure, medetomidine was reversed with atipamezole 0.05 mg/kg i/m. Single dose of methadone (Heptanon, Pliva, Zagreb, Croatia) 0.4 mg/kg s/c was administered for postoperative analgesia. Panting that was observed soon thereafter was thought to be a consequence of methadone administration or relatively warm recovery room (23 to 24°C). The dog's body temperature was not measured because it's aggressive temper. Two hours later, the dog was discharged to a home care with carprofen 2 mg/ kg p/o q12h to be given following three days. At that time, it was alert but still mildly ataxic and weak. Later in the afternoon, the dog's caretaker noticed that the dog is apathetic and sleepy, and it refused water and food. Its pulse was thready. The dog was given additional dose of atipamezole (dose not known) by staff veterinarian and sent back to CSAMS-VFLJ. On the admission (7 hours after the end of procedure) the dog was in cardiopulmonary arrest. Cardiopulmonary resuscitation (CPR) was performed with no success. During CPR, considerable amount of smelly dark reddish intestinal contents were released. Post-mortem examination carried out at Institute for Pathology, Forensic and Administrative Veterinary Medicine, Veterinary Faculty, University of Ljubljana revealed severely congested kidneys, liver and small intestinal wall. The small intestine was filled with hemorrhagic contents, while large intestine was empty. The spleen and lungs were congested as well. The subcutaneous blood vessels were congested with non-coagulated blood and the muscles were pale. Non-coagulated blood was found in right ventricle. Nosocomial Klebsiella oxytoca infection in two dogs 119 Histopathology showed disintegrated epithelium of small intestine. Small intestinal mucosal venous dilatation was accompanied with lymphocyte and macrophage infiltration in the lamina propria. Inflammatory cells, predominantly lymphocytes, were present in liver vessels. Liver sinusoids were congested and hepatocellular atrophia was observed in centrilobular zone. In both kidneys, hem-orrhagic renal cortical necrosis with tubular casts was found. Reactive hyperplasia of spleen involved lymphoid tissue of the white pulp and macrophages of the red pulp. Several small colonies of bacteria were found in blood vessels and in parenchyma-tous tissue of the spleen. Severe congestion with multiple focal intraalveolar haemorrhages was found in bilateral lungs, and a small number of alveoli contained air. In the heart, prominent multiple epicardial haemorrhages and myocardial hy-peraemia were observed. Abdominal organs were sent to bacteriological examinations and Klebsiella oxytoca was isolated abundantly from liver, kidneys and spleen. No bacteria were isolated from lungs. Klebsiella oxytoca and haemolytic Escherichia coli were isolated from small intestine while Klebsiella oxytoca and Clostridium sporogenes were isolated from the small amount of peritoneal fluid, the latter probably contaminating abdominal cavity post mortem. Initial tests of isolated bacteria regarding beta-lactamase activity by nitrocefin were strongly positive (Table 1). Screening tests for extended-specter-p-lactamase (ESBL) were negative and confirmation tests with E-test were negative as well. The antibiotic susceptibility of isolated strains of Klebsiella oxytoca and Escherichia coli was determined as in dog 1 (Table 1). The anaesthesia staff was concerned about a possible common-source epidemic of Klebsiella oxytoca infection after the second case occurred, and environmental surfaces and equipment in pre-/post-operative rooms (PPR), operating theatres and wards (tables, cages, incubator, clippers, muzzles, Doppler pulse detector and pulse oximetry probes, breathing circuits, endotracheal tubes, laryngoscope blades, dentistry equipment) as well as opened vials of propofol, acepro-mazine and carprofen and intravenous fluids were bacteriologicaly examined. p-lactamase negative Klebsiella oxytoca (by nitrocefin testing) was isolated from PPR cages and Staphylococcus intermedius, Entero-coccus sp., and Bacillus sp. from muzzles. p-lactamase positive Klebsiella oxytoca and p-lactamase positive Escherichia coli (by nitrocefin testing) were isolated from tables, cages and clippers in wards. Additional infection control measures for PPR, operating theatres and wards were instituted, including restrictions on the personnel entrance into PPR and operating theatres, strict disinfection of clippers, tables, cages and other equipment between the patients and regular disinfection of hands before handling another animal or before intravenous, arterial and urinary catheter placement. Particular care was taken at manipulation of catheters and disconnection of infusion lines. The use of single-use gloves was strongly encouraged when manipulating the animals. So far, 24-months after the additional infection control measures were instituted; no nosocomial infection was confirmed at CSAMS-VFLJ. Discussion 90% of nosocomial bloodstream infections in human hospitals are related to intravenous catheter use (23). These infections can range in severity from localized phlebitis to fatal bacteraemia and sepsis, and are caused by contamination of the catheter either at the time of insertion or during use (24). Microorganisms may be introduced on the hands of the person placing or handling the catheter. They may be a part of the health care worker's own skin flora, may be part of the patient's skin flora acquired from a site that has not been disinfected, or may be faecal or other types of bacteria that the health care worker has handled (1). There are few data in the veterinary literature documenting the incidence of catheter-related bloodstream infections in companion animals (2, 4, 5, 6, 7, 9, 10, 11, 12, 13). Most catheter-related infections involve organisms that are ubiquitous to the patient's skin flora, nasopharynx or intestinal tract and organisms carried on the hands of hospital personnel (25). Surveillance study in a veterinary hospital reported 26% positive jugular catheter bacterial cultures after an average of 2.7 days. More than 50% of the organisms were Klebsiella spp. and En-terobacter spp (1). In the present report, the cages and clipper from wards were contaminated with p-lactamase positive Klebsiella oxytoca, which was also isolated from both affected dogs. None of dogs entered the wards; therefore the only possible way of transmission could be by the hands of personnel rotating between PPR and wards or the clipper from the wards although the rotation of clipper between the wards and PPR is not a common practice at CSAMS-VFLJ. However, since the culturing of microorganisms from environmen- 120 A. Seliskar, I. Zdovc, B. Zorko tal surfaces and equipment was carried out 4 days after the dog 2 died, regular cleaning and disinfection procedures in a meantime might have an influence on the microbiology results and other sources of intravenous catheter contamination can not be excluded. While there is the possibility that the infection entered the bloodstream at the oral cavity during dental procedure in dog 2, it is highly unlikely in dog 1 because during the implantation of bilateral direct acrylic plane the integrity of gingival mucosa was not interrupted. However, the lack of catheter cultures makes it hard to be certain of the source of the infection. Clinical presentation of Klebsiella spp. bacterae-mia is indistinguishable from that of bacteraemia caused by other microorganisms. If bacteraemia develops, symptoms include fever or hypothermia, leukocytosis with left shift or neutropenia, and shock. The incidence of septic shock is remarkable; in a five-year study in human hospital 22% of the patients with documented bacteraemia due to Klebsiella spp. developed septic shock (26). In the present report, shock was overlooked in a dog 2, because the dog was discharged soon after the end of procedure due to his aggressive temper and uncoop-erativeness, and the early signs of impending shock such as panting and change in mental status were unfortunately misinterpreted. The results of post-mortem examination of dog 2 indicate that the dog died because of septic shock associated with disseminated intravascular coagulation, and p-lactamase positive Klebsiella oxytoca was isolated from several abdominal organs, including small intestine. The authors can not affirm, whether Klebsiella oxytoca was introduced into the dog through intravenous catheter or during dental procedure or the dog was already the reservoir for Klebsiella oxytoca, which spread to extraintestinal sites through mesenteric lymph node complex as a result of bacterial translocation. Bacterial translocation in hosts with an intact intestinal barrier can occur by intracellular route, however the primary mechanism promoting bacterial translocation in these cases is bacterial overgrowth due to recent antimicrobial therapy (27), which was not the case in dog 2. The other mechanism, which promotes bacterial translocation is increased permeability of intestinal mucosal barrier, for example due to hem-orrhagic or endotoxic shock, where indigenous bacteria translocate intercellulary (28). The various species of indigenous bacteria do not all translocate at the same rate. Gram-negative, facultative anaerobic Enterobacteriaceae, such as Escherichia coli, Klebsiella spp., and Proteus mirabilis, translocate from gastrointestinal tract at a greater rate than the other bacteria (27). In the present report, the authors doubt that uneventful short-term anaesthesia could provoke bacterial translocation and sepsis by itself in previously healthy dog. The susceptibility profile of Klebsiella oxytoca isolated from dog 1 and dog 2 showed that the organism was not resistant to a broad spectrum of antibiotics including those commonly used in the hospital (amoxicillin+clavulanic acid, gentamicin, enrofloxacin). Dog 2's antimicrobial sensitivity profile showed an intermediate resistance to cephalex-in, where dogl's organism was sensitive. This might represent a developing and broadening resistance pattern as the organism has been presumably in a hospital setting for a longer period of time, but on the other hand cephalexin is not an antibiotic used on a regular basis in a hospital setting. Amoxicillin+clavulanic acid to which Klebsiella oxytoca was sensitive, is used most often for prophylactic antibiotic treatment of surgical and dentistry patients at CSAMS-VFLJ. The lipopolysaccharide capsule of Klebsiella oxytoca that protects the organism from phagocytosis and aids in adherence (21) might be the cause for the severity and rapid development of the infection in dog 2. In conclusion, the present report demonstrates the potential for nosocomial infection with Klebsiel-la oxytoca in small animal hospitals. Strict infection control measures, particularly regular disinfection of hands and use of disposable gloves should be strongly encouraged as the most important factors in preventing nosocomial infections. References 1. 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Infect Immun 1987; 55 (1): 44-8. 22. Gniadkowski M. Evolution and epidemiology of extended spectrum ß-lactamases (ESBLs) and ESBL-producing microorganisms. Clin Microbiol Inf 1991; 7: 597-608. 23. Clemence MA, Walker D, Farr BM. Central venous catheter practices: results of a survey. Am J Infect Control 1995; 23: 5-12. 24. Bjornson HS. Pathogenesis, prevention, and management of catheter-associated infections. New Horiz 1993; 1: 271-8. 25. Spurlock SL, Spurlock GH. Risk factors of catheter-related complication. Compend Contin Educ Pract Vet 1990; 12: 241-8. 26. Garcia de la Torre MG, Romero-Vivas J, Martinez-Beltran J, et al. Klebsiella bacteremia: an analysis of 100 episodes. Rev Infect Dis 1985; 7: 143-50. 27. Berg RD. Bacterial translocation from the gastrointestinal tract. In: Paul PS, Francis DH, eds. Mechanisms in the pathogenesis of enteric diseases. New York: Kluwer Academic/Plenum Publishers, 1999: 11-30. 28. Berg RD. The immune response to indigenous intestinal bacteria. In: Hentges D, ed. The intestinal microflora in health and disease. New York: Academic Press, 1983: 101-26. 122 A. Seliskar, I. Zdovc, B. Zorko BOLNIŠNIČNA OKUŽBA DVEH PSOV Z BAKTERIJO KLEBSIELLA OXYTOCA A. Seliškar, I. Zdovc, B. Zorko Povzetek: Kljub številnim literaturnim podatkom o bolnišničnih okužbah prek intravenskega katetra z bakterijo Klebsiella oxytoca v humani medicini so podatki s področja veterinarske medicine pomanjkljivi. Predstavljena sta dva primera suma okuženosti intravenskega katetra z bakterijo Klebsiella oxytoca pri psih, pri katerih je bil opravljen stomatološki poseg v splošni anesteziji. Pri prvem psu se je 3 tedne po vstavitvi obojestranske direktne akrilatne opornice razvil osteomielitis ramenskega sklepa na nogi, ki smo jo ob posegu uporabili za intravensko kateteriza-cijo. Bakteriološka identifikacija sklepne tekočine je potrdila okužbo z bakterijo Klebsiella oxytoca, psa pa smo glede na rezultate antibiograma mesec dni zdravili z amoksicilinom s klavulansko kislino. Kljub kliničnemu izboljšanju, smo z rentgenskim slikanjem ob zaključku zdravljenja potrdili osteoartritis ramenskega sklepa. Drugi pes je poginil 7 ur po izdrtju levega mandibularnega četrtega ličnika zaradi septičnega šoka z diseminirano intravaskularno koagulacijo. Po smrti smo izolirali bakterijo Klebsiella oxytoca iz več organov trebušne votline. Zaradi suma glede skupnega izvora okužbe smo opravili bakteriološko preiskavo bolnišničnih prostorov in opreme ter med drugim identificirali tudi bakterijo Klebsiella oxytoca. Poostrili smo postopke za preprečevanje prenosa morebitnih bolnišničnih okužb in 18 mesecev po opisanih primerih nismo potrdili novih bolnišničnih okužb. Ključne besede: bolnišnična okužba - mikrobiologija; infekcija, nadzor; Klebsiella infekcije - etiologija - mikrobiologija; pooperacijske komplikacije; psi Slov Vet Res 2007; 44 (4): 123-32 UDC 636.3.09:616.98-07:616-036.22:551.585.53 Case Report LARGE SCALE OUTBREAKS OF PESTE DES PETITS RUMINANTS IN SHEEP AND GOATS IN THAR DESERT OF INDIA Anil Kumar Kataria1*, Nalini Kataria2, Ajey Kumar Gahlot3 1Apex Centre for Animal Disease Investigation, Monitoring and Surveillance, 2Department of Veterinary Physiology, Radioisotope Laboratory, 3Department of Clinical Veterinary Medicine, College of Veterinary and Animal Science, Rajasthan Agriculture University, Bikan-er-334001, Rajasthan, India Corresponding author, E-mail: akkataria1@rediffmail.com Summary: Severe peste des petits ruminants (PPR) outbreaks were recorded in Thar desert of Rajasthan state (India) affecting large population of sheep and goats over a huge geographical area, taking a toll in tens of thousands of animals. These outbreaks were investigated for epidemiological, clinical, pathological, haematological and biochemical features. The outbreaks started in migratory sheep which returned back from adjacent PPR endemic areas and later spread to goats with estimated case fatality rates of 70% and 80%, respectively. Younger animals were more affected than the adult ones. The stress of migration coupled with low environmental temperatures bolstered by humidity and nutritional deficiency was the reason of precipitation of the disease. Typical clinical and pathological findings were recorded and significant (P^0.05) changes were observed in most of the haemato-biochemical parameters. The free-range grazing system had been responsible for the involvements of larger number of flocks in distant areas with prevalence for longer duration. The disease waned with increase in the environmental temperatures. The present situation warrants strict sero-surveillance and monitoring of PPR together with uninterrupted vaccination of migratory flocks at district or state borders for its effective control. Keywords: sheep diseases - diagnosis - virology; goat diseases - diagnosis - virology; disease outbreaks; peste-despetits-ruminants - diagnosis - virology; pathology, clinical; blood chemical analysis: antibodies, viral - blood; enzyme-linked immunosorbent assay - methods; desert climate; sheep; goats Introduction Peste des petits ruminants (PPR), an office international des epizooties list A disease of sheep and goats caused by morbillivirus is characterized by high morbidity and high mortality rates resulting into heavy economical losses. The disease has been regularly reported from various parts of the world but mainly from Africa, Arabia, the Middle and Near East and the Indian subcontinent (1, 2, 3, 4). Rajasthan state situated in the north-west part of India is the biggest state in regards to its geographical area and shares a very long international border with Pakistan. The inland borders are shared with other states: Punjab in the north, Haryana and Uttar Pradesh in the north-east, Madhya Received: 1 October 2007 Accepted for publication: 19 November 2007 Pradesh in the south-east and Gujarat in the southwest. The arid region of western part of Rajasthan constitutes famous Thar desert, where ambient temperatures vary widely from subzero in winter to as high as 50 °C in summer in some places. In this area, the livestock remains the mainstay of desert economy over agricultural produce. The sheep are reared mainly for wool and meat, and goats for milk and meat. The migration of small ruminants, especially of sheep, to other adjoining states, is a regular feature during famine and during times of scarcity of feed and fodder in Thar desert. The migration of various sized flocks (usually of few hundreds to several thousands) normally starts in March which is the time of onset of summer in this region. The flocks return back by October, the time of onset of winter, but early return of animals is possible if rain is experienced in the area. The arrival of animals is often accompanied and/or followed by various dis- 124 A. K. Kataria, N. Kataria, A. K. Gahlot ease outbreaks in the unprotected flocks. Though PPR is considered endemic throughout India (5) this region had never experienced the outbreaks in the past similar to that in the present report involving a wider geographical area with very high morbidity and mortality. This paper puts on record some field investigations in regards to epidemiology, clinical signs, pathological findings, and laboratory investigations encompassing haemato-biochemical changes and histopathology in sheep and goats affected with PPR during natural outbreaks in the Thar desert. Material and methods A series of PPR outbreaks were observed in the northwest Rajasthan affecting both sheep and goat populations between November 2005 and April 2006. Sheep were mostly Magra, Marwari and Rambouillet breeds and their crosses with local breeds in the organized farms and goats belonged to Mar-wari breed. The epidemiological observations were recorded in several outbreaks in 21 villages and one organized sheep breeding farm. The clinical signs were observed and post-mortem examination of 58 carcasses was carried out at various places. The blood samples were drawn from 30 sheep (15 adults and 15 lambs) and 30 goats (15 adults and 15 kids) affected with PPR and 20 healthy sheep (10 adults and 10 lambs) and 20 healthy goats (10 adults and 10 kids) for haemato-biochemical analyses. The most of the adult animals were females but young ones included both male and female. Various haemato-biochemical parameters were determined by standard techniques (6, 7, 8) and serum cortisol by radioimmunometric method using 125I radioim-munoassay kit (RADIM, Spain). The statistical significance for a given parameter was determined between healthy and PPR affected animals. Comparison between healthy and infected herds was made by student's t-test with significant difference considered at p < 0.05 (9). Results Field Investigation Epidemiology The disease first appeared in November 2005 in migratory sheep flocks in Sardarshahar area of Churu district adjoining to Haryana state and later spread to other adjoining districts. The widespread outbreaks were encountered in different flocks until April 2006. The highest morbidity and mortality rates were recorded during the periods when the lowest night temperatures dropped below zero in January 2006 with high humidity. Because of involvement of very large animal population in huge geographical area in the desert, the exact figures could not be gathered but the available data revealed death of few thousand animals with a case fatality of 70% in sheep and 80% in goats. The young animals of both species had higher case fatality rates than their respective adults. The disease was also recorded at an organized sheep farm where mortality was observed mostly in young animals below 3 months of age which were not vaccinated against PPR, however, vaccinated adult animals also died albeit with less severe clinical and pathological findings. Clinical signs The course of the disease was acute and suba-cute, few of the animals died even in 36 hours of onset of the disease. The affected animals initially were severely depressed with a sudden rise in body temperature reaching almost 42 °C in some cases, and the fever persisted for 7-8 days. From the onset of fever, most animals had a serous nasal discharge which progressively turned into mucopurulent discharge, leading to severe respiratory distress. Areas of erosions were most commonly seen on the visible nasal mucous membranes and muco-cutaneous junctions with inflammation around the mouth (Fig.1). In many of the animals, lesions similar to orf developed at mucocutaneous junction of mouth. The erosive and necrotic stomatitis started as areas of hyperemia at gums, cheeks, dental pad and / or anterior dorsal part of tongue with frothy salivation. The areas later developed into irregular non-haemor-rhagic lesions (Fig.2) and in some of the cases circular raised but flat non-bleeding lesions were present on the tongue (Fig.3). There was a great amount of necrotic debris on the older lesions (Fig.4). The individuals with severe oral lesions had visible swelling around mouth. A non-haemorrhagic diarrhoea was observed in all affected animals, developing 2-3 days after onset of the disease. Conjunctivitis was recorded with lachrymal discharge which became mucoid resulting in sticky eyelids. Abortion in pregnant animals was a consistent feature and vulvar mucous membranes had erosive lesions very similar to that in intestinal mucosa.. A subnormal temperature preceded death in animals with severe diarrhoea for few days. Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert of India 125 Figure 1: Erosions at muco-cutaneous junction with inflammation around the mouth (PPR kid) Figure 2: Irregular non-haemorrhagic oral lesions (PPR, sheep) Figure 3: Raised and flat circular lesions on tongue (PPR, goat) Figure 4: Necrotic debris on oral lesions (PPR, sheep) Pathological findings Grossly, the carcasses were dehydrated with apparent swelling around the mouth. The lesions in mouth were consistently present in all animals except those who died within short period of 1-2 days. The lesions of respiratory tract included necrotic areas on the mucosa of nostrils and turbinates, severely congested tracheal mucous membrane and white froth in trachea. Lungs were congested and consolidated especially involving antero-ventral parts. Hydrothorax was recorded in few cases. The oral cavity was full of white necrotic debris where oral lesions were severe. The lesions in gastrointestinal tract consisted of few erosions in the mucosa from where blood oozed out, large intestines were congested, especially at the caeco-colic junction with streaks of blood on mucosal crests (Zebra-stripes) (Fig.5) though the zebra-stripes were not seen in all the carcasses. The lymph nodes, especially from mesenteries were severely oedematous, congested and enlarged (Fig.6). In some cases haemorrhages on the internal walls of gall bladder were recorded (Fig.7). Spleen was slightly enlarged in few cases. In some animals liver was studded with necrotic foci. Vaginitis was observed in many animals and vulvar mucus membranes were inflamed and had erosive lesions. Though the types of lesions were more or less similar in all the animals, there was a variation in the severity and involvement of the organs. The vaccinated animals had less severe lesions on tongue. Laboratory Investigations Haemato-biochemical parameters The mean values of haemato-biochemical parameters in healthy animals and PPR affected sheep and 126 A. K. Kataria, N. Kataria, A. K. Gahlot Figure 5: Zebra-stripes in large intestine (PPR sheep) Figure 6: Oedematous, congested mesenteric lymph node (PPR, goat) Figure 7: Haemorrhages on the internal wall of gall bladder (PPR, sheep) goats are presented in table 1 through 4. All parameters included in the investigation significantly differed between healthy and infected animals in both species in both young and adult animals (p<0.05): total erythrocyte count (TEC), total leucocyte count (TLC), haemoglobin (Hb), packed cell volume (PCV), differential leucocyte count (DLC), relative viscosity, specific gravity and erythrocytic sedimentation rate (ESR). In PPR affected animals of both species and age groups significant (p<0.05) changes were recorded in serum biochemical parameters for sodium, potassium, glucose, total serum proteins, albumin, globulin, creatinine, urea and cortisol. Histopathological changes Histopathologically in lungs syncytial cells were present in alveolar lamina and inclusion bodies in cytoplasm and nucleus of alveolar macrophages and epithelial cells of bronchioli and bronchi. There was severe congestion of mucosa and submucosa and degeneration and necrosis of intestinal epithelium. Serology The sera collected from representative animals in different areas who survived the disease were subjected to c-ELISA which confirmed the presence of specific antibodies. Discussion Field Investigations Epidemiology The disease first appeared in the sheep flocks which returned back after migration to other states during periods of famine in this desert area. The arrival of flocks was coincided with sudden lowered ambient temperatures. It is expected that the sheep on migration to other states came in contact with native small ruminant population where endemicity of the disease is much higher. The stress of migration and environmental temperature might have precipitated the onset of disease (10). The animals during feed scarcity often become nutritionally deficient resulting into increased susceptibility to infection (5). The spread of disease to other adjoining districts was due to unrestricted movements of the animals as they are reared on free range system of feeding and management covering long distances during the day (10, 11). The serial occurrence of dis- Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert of India 127 Table 1: Haematological values (Mean ± SEM) in healthy and PPR affected sheep S. Sheep No. Parameter Adults Lambs Healthy PPR affected Healthy PPR affected (n = 10) (n = 15) (n = 10) (n = 15) 1. TEC (x1012/ l) 7.92+0.23 8.99+0.31* 8.91+0.41 10.0+0.39* 2. TLC (x109/ l) 9.98+0.31 6.0+0.37* 11.11+0.22 7.11+0.41* 3. Hb (g/dl) 10.41+0.61 13.11+0.31* 13.34+0.51 15.12+0.31* 4. PCV (%) 29.33+0.61 34.12+0.39* 34.12+0.58 37.22+0.34* 5. DLC (%) Neutrophil 35.3+0.30 61.8+0.30* 36.3+0.22 60.9+0.25* Lymphocyte 55.2+0.21 35.2+0.21* 54.8+0.31 35.8+0.20* Monocyte 5.3+0.05 2.3+0.01* 5.4+0.01 1.9+0.01* Eosinophil 3.1+0.03 1.1+0.01* 3.0+0.01 1.1+0.01* Basophil 0.6+0.02 0.2+0.001* 0.5+0.001 0.15+0.001* 6. Relative Viscosity 4.3+0.03 4.9+0.04* 4.6+0.05 5.0+0.03* 7. Specific gravity 1.050+0.003 1.059+0.001* 1.055+0.002 1.060+0.001* 8. ESR (mm/4Hr.) 1.93+0.04 0.20+0.04* 1.48+0.02 0.30+0.03* 1. Figures in parenthesis indicate number of animals. 4. TEC = Total erythrocyte count 2. In adults and lambs mean comparison has been made 5. TLC ^ = Total leucocyte count between respective healthy and PPR affected animals 6. Hb = Haemoglobin for each parameter. All the differences were significant 7. PCV= = Packed cell volume (p<0.05) as indicated by asterix. 8. DLC= = Differential leucocyte count 3. PPR = Peste des petits ruminants 9. ESR = Erythrocytic sedimentation rate Table 2: Biochemical parameters (Mean ± SEM) in healthy and PPR affected sheep S. Sheep No. Parameter Adult Lambs Healthy PPR affected Healthy PPR affected (n = 10) (n = 15) (n = 10) (n = 15) 1. Sodium (mmol/L) 130.11+1.89 140.12+2.00* 133.2+1.34 144.2+3.0* 2. Potassium(mmol/L) 5.3+0.1 8.0+0.10* 5.5+0.10 9.12+0.09* 3. Calcium(mmol/L) 2.40+0.025 2.25+0.022NS 2.45+0.022 2.32+0.017NS 4. Phosphorus(g/L) 0.043+0.0034 0.040+0.0007NS 0.045+0.0007 0.043+0.0003NS 5. Glucose (mmol/L) 2.48+0.072 1.65+0.055* 2.56+0.055 1.54+0.049* 6. TSP ( g/L ) 71.2+1.0 61.0+3.1* 78.0+1.1 63.0+ 1.1* 7. Albumin( g/L ) 39.4+1.2 23.1+0.9* 41.2+0.9 23.8+0.9* 8. Globulin( g/L) 36.0+0.9 36.9+0.8* 36.8+0.9 40.1+0.7* 9. Creatinine(|umol/l) 97.24+3.53 185.64+2.65* 79.56+2.65 185.64+1.76* 10. Urea(mmol/L) 3.83+0.018 4.99+0.021* 3.33+0.021 4.69+0.019* 11. Cortisol(mmol/L) 0.165+0.011 0.259+0.019* 0.173+0.008 0.312+0.019* 1. Figures in parenthesis indicate number of animals. 2. In adults and lambs mean comparison has been made between respective healthy and PPR affected animals for each parameter. All the parameters except calcium and phosphorus showed significant differences (p<0.05) as indicated by asterix. 3. NS= Non significant difference (p>0.05) 4. PPR = Peste des petits ruminants 5. TSP = Total serum proteins 6. NS = Non-significant 128 A. K. Kataria, N. Kataria, A. K. Gahlot Table 3: Haematological values (Mean ± SEM) in healthy and PPR affected goats S. No. Parameter Goats Adults Kids Healthy (n = 10) PPR affected (n = 15) Healthy (n = 10) PPR affected (n = 15) 1. TEC (x1012/ l) 8.22+0.41 0.11+0.51* 9.00+0.38 11.31+0.39* 2. TLC (x109/ l) 9.31+0.81 5.93+0.38* 10.99+0.31 6.43+0.22* 3. Hb (g/dl) 10.12+0.31 13.12+0.41* 11.88+0.21 13.88+0.34* 4. PCV (%) 28.22+1.0 33.12+0.91* 32.34+1.01 37.12+0.93* 5. DLC Neutrophil Lymphocyte Monocyte Eosinophil Basophil 36.1+0.12 55.3+0.16 5.1+0.03 3.0+0.04 0.5+0.001 62.4+0.21* 34.2+0.20* 2.1+0.02* 1.0+0.01* 0.1+0.001* 37.0+0.22 54.0+0.15 5.0+0.01 3.0+0.01 0.4+0.001 66.3+0.31* 32.8+0.41* 2.10+0.01* 1.12+0.01* 0.15+0.001* 6. Relative Viscosity 4.0+0.04 4.8+0.03* 4.25+0.06 4.9+0.02* 7. Specific gravity 1.050+0.001 1.055+0.001* 1.054+0.001 1.059+0.001* 8. ESR (mm/4Hr.) 1.4+0.03 0.27+0.03* 1.53+0.04 0.30+0.04* 1. Figures in parenthesis indicate number of animals. 4. TEC = Total erythrocyte count 2. In adults and lambs mean comparison has been made 5. TLC = Total leucocyte count between respective healthy and PPR affected animals 6. Hb = Haemoglobin for each parameter. All the differences were significant 7. PCV= Packed cell volume (p<0.05) as indicated by asterix. 8. DLC= = Differential leucocyte count 3. PPR = Peste des petits ruminants 9. ESR = Erythrocytic sedimentation rate Table 4: Biochemical parameters (Mean ± SEM) in healthy and PPR affected goats S. No. Parameter Goats Adults Kids Healthy (n = 10) PPR affected (n = 15) Healthy (n = 10) PPR affected (n = 15) 1. Sodium (mmol/L) 131.12+1.31 141.1+0.71* 133.0+1.7 145.2+0.61* 2. Potassium(mmol/ L) 5.6+0.09 7.9+0.05* 5.9+0.09 8.1+0.13* 3. Calcium(mmol/ L) 2.46+0.032 2.42+0.005NS 2.475+0.05 2.42+0.05NS 4. Phosphorus(g/L) 0.0583+0.0004 0.056+0.0009 NS 0.057+0.0009 0.055+0.0009NS 5. Glucose (mmol/L) 2.45+0.044 2.101+0.038* 2.59+0.038 2.15+0.028* 6. TSP ( g/L ) 70.7+0.9 60.0+1.0* 77.1 + 1.0 61.0+ 0.2* 7. Albumin( g/L ) 36.2+0.1 21.0+0.3* 36.3+0.3 22.0+ 0.3* 8. Globulin( g/L) 34.1+0.1 39.0+0.6* 34.3+0.2 38.0+0.8* 9. Creatinine(|umol/l) 97.24+7.70 167.96+3.53* 114.92+2.65 238.68+7.95* 10. Urea(mmol/L) 2.49+0.049 3.83+0.019* 2.05+0.066 3.33+0.0166* 11. Cortisol(mmol/L) 0.168+0.009 0.284+0.022* 0.165+0.008 0.259+0.008* 1. Figures in parenthesis indicate number of animals. 2. In adults and lambs mean comparison has been made between respective healthy and PPR affected animals for each parameter. All the parameters except calcium and phosphorus showed significant difference (p<0.05) as indicated by asterix. 3. NS= Non significant difference (p>0.05) 4. PPR = Peste des petits ruminants 5. TSP = Total serum proteins 6. NS = Non-significant Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert of India 129 ease outbreaks had also been recorded for a long duration in different flocks in various adjoining districts in west Bengal (10). The highest morbidity and mortality rates were recorded during the periods when the lowest night temperatures dipped below zero in January 2006 with high humidity. In the present outbreaks humidity might have played the role in disease occurrence as it may be one of the factors allowing thriving and multiplication of PPR virus in nature (10). The environmental stress particularly hot and humid is also held responsible for precipitation of the disease (12). Because of involvement of very large animal population in huge geographical area in desert the all figures could not be gathered regarding morbidity and mortality nevertheless the available data suggested a case fatality of 70% in sheep and 80% in goats. The PPR antibody prevalence is low (10- 30%) in goat population in the northern parts of the country suggesting higher risk of infection in goats (5). Moreover, the recovery rate of goats infected with PPR virus is less in comparison to that in sheep (13). In the Thar desert sheep and goats are reared in unorganized sector by small and marginal farmers and these animals are used as cash crops without any inputs. Neither these animals are supplemented with feed additives nor usually vaccinated against the diseases in which cost is incurred. This area never experienced PPR before hence all of the animals in these outbreaks were also not protected against it. The non-vaccination of these animals explains the very high morbidity and mortality rates. In the present serial outbreaks both species of small ruminants were affected whereas in some earlier reported outbreaks only goats died and sheep were not affected at all (10, 14). In contrast to the present observation a significantly higher mortality rate was recorded in sheep than in goats in Cameroon (15). The disease was also recorded at an organized sheep farm where mortality was observed mostly in young animals below 3 months of age which were not vaccinated against PPR. It has been suggested that new born animals become susceptible to PPR virus infection at three to four months of age (16). However, few vaccinated adult animals also died albeit with less severe clinical and pathological findings. Clinical signs Though the course of the disease was acute and subacute with appearance of clinical signs and development of typical lesions, in cases where animals died within 1-2 days of onset of disease no lesions were recorded. Most of the affected animals had very high body temperatures for long periods. The circular raised, but flat lesions on dorsum of tongue were seen only in few cases. The development of lesions on muco-cutaneous junction of mouth very similar to that in orf or contagious ecthyma needs to be differentiated. The presence of mucopurulent discharge and froth in trachea led to respiratory occlusion and development of a typical obstructive sound during breathing. The amount of necrotic debris in oral cavity depended on the extent and age of the lesions. Unlike in other outbreaks typical overt oral lesions were recorded in the present outbreaks in sheep as well as in goats (17). A continuous profuge diarrhoea caused severe dehydration in the affected animals. The kids were more severely affected due to dehydration and showed prostration for longer duration as compared to lambs. These animals had more subnormal temperatures before death. Pathological findings The mouth lesions were present in all of the animals except those who died within 1-2 days of appearance of disease. Even the vaccinated animals at the organized sheep breeding farm had lesions on tongue. Lungs in all the animals showed congestion and consolidation but hydrothorax was recorded in few cases only. No animal had any kind of lesion in abomasums (17) and heavy haemorrhages in large intestines (18) were not observed in any of the animals. The haemorrhages recorded on the luminal wall of gall bladder in the present outbreak in some cases has not been reported to the best of our knowledge but presence of thick granular bile had been reported (14). The necrotic foci in liver of some animals could have developed due to some secondary bacterial infection of haematogenous route. Though the types of lesions were more or less similar in all the animals but there was variation in the severity and involvement of the organs. Treatment The affected animals were given antibiotics to control secondary bacterial infections along with anti-inflammatory drugs. Many animals which received proper treatment in early stages of the disease were saved. It was observed that higher fatality rates in PPR affected animals was more due to secondary bacterial infections than the disease itself 130 A. K. Kataria, N. Kataria, A. K. Gahlot which could have been due to immune suppression associated with morbillivirus infections (19). Laboratory Investigations Haemato-biochemical parameters The mean values of haemato-biochemical parameters in healthy animals corroborated the earlier findings in sheep and goats of arid tract (20, 21, 22). The severe dehydration in the affected animals was evidenced by increased viscosity and specific gravity which led to polycythaemia (23). Severe leu-copoenia could have been due to the inhibition of peripheral blood lymphocytes proliferation by PPR virus (19). A marked lymphocytopoenia, monocyto-poenia, neutrophilia and eosinopoenia in present investigation could have been due to the combined effect of virus infection and stress as evidenced by elevated cortisol levels (24). The increased mean values of sodium and potassium reflected haemoconcentration. The total serum protein values decreased but globulin concentration increased indicating immune response towards infection. The higher globulin concentration was achieved at the expense of compensatory fall in albumin levels. Serum cortisol was recorded higher in PPR affected stock indicating stress. Cortisol causes increase in blood glucose levels due to glycogenolytic property but in present investigation, decreased glucose levels could have been due to animals not being fed since the onset of infection. The higher levels of cortisol causing muscle wasting resulted in increased serum creatinine levels and an increased urea concentration reflected protein breakdown and haemo-concentration simultaneously. Acknowledgements The authors are thankful to Joint Director, CADRAD, Indian Veterinary Research Institute, Bareilly (UP) for serological confirmation of the disease. References 1. Obi TU, Durojaiye OA, Kasali OB, Akpavie S, Opasina, DB. Peste des petits ruminants (PPR) in goats in Nigeria: clinical, microbiological and pathological features. Zentralbl Vet Med B 1983; 30: 751-61. 2. Al-Naeem A, Elzein EMA, Al-Afaleq AI Epizootiologi-cal aspects of peste des petits ruminants and rinderpest in sheep and goats in Saudi Arabia. Rev Sci Tech Off Int Epizoot 2000; 19: 855-8. 3. Taylor WP, Al-Busaidy S, Barrett T. The epidemiology of peste des petits ruminants in the Sultanate of Oman. Vet Microbiol 1990; 22: 341-52. 4. Shaila MS, Purushothaman V, Bhavsari D, Venugopal K, Venketesan RA. 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Epidemiologi-cal observations on Peste des petitis ruminants in north India. Indian J Anim Sci 1999; 69: 365-8. 12. Wosu LO. Seasonality of occurrence of peste des petits ruminats disease in Nigeria: a preliminary report. Mysore J Agric Sci 1995; 29: 158-62. 13. Shankar H, Gupta VK, Singh N. Occurrence of peste des petits ruminants like disease in small ruminants in Uttar Pradesh. Indian J Anim Sci 1998; 68: 38-40. 14. Shaila MS, Peter AB, Varalakshmi P, Apte M, Ra-jendran MP, Anbumani SP. Peste des petits ruminants in Tamilnadu goats. Indian Vet J 1996; 73: 587-8. 15. Ndamukong KJ, Sewell MM, Asanji MF. Disease and mortality in small ruminants in the North West Province of Cameroon. Trop Anim Health Prod 1988; 21: 191-6. 16. Srinivas RP, Gopal T. Peste des petits ruminants (PPR): a new menace to sheep and goats. Livestock Advisor 1996; 21: 22-6. 17. Kumar A, Singh SV, Rana R, Vaid RK, Misri J, Vihan VS. PPR outbreak in goats: epidemiological and therapeutic studies. Indian J Anim Sci 2001; 71: 815-8. 18. Singh VP, Chum VK, Mondhe KS. Peste des petits ruminants: an outbreak in sheep in Rajasthan. Indian Vet J 1996; 73: 466-7. 19. Heaney J, Barrett T, Cosby SL. Inhibition of in vitro leukocyte proliferation by morbilliviruses. J Virol 2002; 76: 3579-84. 20. Kataria AK, Kataria N, Gahlot AK. Observations on some blood analytes in Marwari sheep. Vet Pract 2005; 6: 39-40. Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert of India 131 21. Kataria AK, Kataria N, Ghosal AK. Haematological changes associated with season in Marwari goats. Indian Vet J 1992; 69: 552-4. 22. Kataria AK, Kataria N, Bhatia JS, Ghosal AK. Blood metabolic profile of Marwari goats in relation to seasons. Indian Vet J 1993; 70: 761-2. 23. Bhikane AU, Kulkarni DD, Narladkar BW, Ali MS. Clinico-pathological and therapeutic investigations on peste des petits ruminants in goats. Indian Vet J 1997; 74: 377-9. 24. Kataria N, Kataria AK. Use of blood analytes in assessment of stress due to drought in camel. J Camel Pract Res 2004; 11: 129-33. 132 A. K. Kataria, N. Kataria, A. K. Gahlot OBSEŽNI IZBRUHI KUGE MALIH PREŽVEKOVALCEV PRI OVCAH IN KOZAH V PUŠČAVI THAR V INDIJI A. K. Kataria, N. Kataria, A. K. Gahlot Povzetek: V puščavi Thar v Radžastanu (Indija) smo naleteli na resne izbruhe kuge malih prežvekovalcev (PPR). Bolezen je prizadela populacijo ovac in koz, ki živi na obsežnem geografskem področju, in je pokončala več deset tisoč živali. Izbruhe bolezni smo analizirali iz epidemiološkega, kliničnega, patološkega, hematološkega in biokemičnega vidika. Izbruhi so se najprej pojavili v nomadskih čredah ovac, ki so se vračale s sosednjih področij z endemično prisotno kugo malih prežvekovalcev, kasneje pa se je razširila še na koze. Ocenjena smrtnost pri ovcah je 70-odstotna in pri kozah 80-odstotna. Mlajše živali so bile bolj prizadete kot odrasle. Naglo širjenje bolezni je pogojeval stres zaradi migracije v kombinaciji z nizkimi temperaturami in visoko vlago v okolju ter pomanjkanjem hrane. Ugotovili smo tipične klinične in patološke najdbe, pri večini hematoloških in biokemičnih parametrov pa smo ugotovili statistično značilne spremembe (PS0.05). Zaradi sistema proste paše je bilo prizadetih veliko čred tudi na oddaljenih predelih, bolezen pa se je tam bistveno dlje zadrževala. Število obolenj se je zmanjševalo v odvisnosti z naraščanjem temperature v okolju. Trenutna situacija v zvezni državi Radžastan zahteva strog nadzor bolezni PPR in serološko pregledovanje v kombinaciji z neprekinjenim cepljenjem nomadskih čred na ogroženih področjih in državnih mejah. Ključne besede: ovce, bolezni - diagnostika - virologija; koze, bolezni - diagnostika - virologija; epidemije; peste-despetits-ruminants - diagnostika - virologija; klinična patologija; kri, kemične analize; protitelesa, virusna - kri; encimsko vezan imunosorbentni test - metode; puščave; ovce; koze Slov Vet Res 2007; 44 (4): 133-4 PROF. DR. MILAN POGAČNIK - DOCTOR HONORIS CAUSA Prof. Dr. Milan Pogačnik (left) receiving honorary doctorate from the rector of Ss. Cyril and Methodius University in Skopje Prof. Dr. Gjorgji Martinovski Not many members of the University of Ljubljana had received the title of honoris causa in the last few years. Therefore, the news that professor Pogačnik, full professor of the veterinary pathology, forensic and administrative veterinary medicine, received this honour on September 26th is much more favourable. The title was given to professor Pogačnik by the University of St. Cyril and Method of Skopje, Macedonia, as the recognition for his merits for the development of the veterinary medicine in the Republic of Macedonia, for his significant contribution to the development of the veterinary pathology, forensic and administrative veterinary medicine, for his important help in international research projects and his influence on the scientific, educational and professional development of the Skopje Faculty of Veterinary Medicine, especially for the renovation of the study programme in accordance with the European standards. The ceremony, which took place in the solemn hall of the Skopje University, was attended by the rector, deans, vice deans, senators and managers of the university institutes, the president of EAEVE (European Association for the Establishments of Veterinary Education) professor Marcel Wanner, members of the executive board and deans of some European veterinary faculties. Thirty six years of professor Pogacnik's work in the field of the veterinary medicine has always been connected with the veterinary pathology, forensic and administrative veterinary medicine and, consequently, with the animal care and protection, veterinary public health and environment protection. Interdisciplinary work, which he devoted his energy to, has been combined with different fields of the veterinary medicine, agriculture, husbandry, food and environment protection, medicine, pharmacy, economy and law. Professor Pogačnik is the principal investigator of the research project Animals' health, environment and safe food, as well as the holder of some target oriented and applied projects in the field of sustainable farming in which he successfully incorporated the interdisciplinary work of professionals from agriculture, forestry and veterinary medicine. The international projects he has conducted enabled Slovenian agricultural and veterinary medicine to establish its place in the Europe and worldwide. The repercussions of his work have resulted in the projects concerning the China Karst cultivation. As the dean of Ljubljana Veterinary Faculty, professor Pogačnik has essentially contributed to the recognition of the faculty at home and abroad. For a long time he was a member of EAEVE executive committee, one of the founders and the president of Vet-Nest (Veterinary Network of European Student and Stuff Transfer) which associates veterinary faculties of Budapest, Brno (Czech Republic), Košice (Slova- 134 kia), Wroclaw (Poland), Zagreb (Croatia) Ljubljana and Vienna (Austria) University of Veterinary Medicine. Moreover, professor Pogačnik conducted the international TEMPUS project, dealing with the veterinary education. He also coordinated the Slovenian part of the survey Vet 2020 which goal was to foresee the necessities of the veterinary profession in the next decade. He is also the Slovenian delegate in OIE (Office International de Epizootie), member of ECVAM scientific committee (European Centre for the Validation of Alternative Methods) and, lately, also the member of EFSA directorate (European Food Safety Authority). Professor Pogačnik has mediated his knowledge and experiences achieved in the scientific, educational, organizational, professional, and management work to other European educational establishments. It is crucial to stress his advisory help to the veterinary faculties in the republics of former Yugoslavia. He has participated as the reviewer in the evaluation of study programmes. As an excellent scientist, lecturer and manager he has actually been the mentor of all faculties in the former common country. This part of the Europe has struggled with many difficulties in the last decade, but the professional connections and the relations have never been cut off. At the ceremony, the dean of the Faculty of the Veterinary Medicine in Skopje professor Mišo Hris-tovski stressed the paramount value of the help given by professor Pogačnik to the Skopje Faculty, especially for its international enforcement. With professor's help, the Faculty of Veterinary Medicine in Skopje, together with Veterinary Faculty in Ljubljana, participates in different international research projects. With the endeavours of profes- sor Pogačnik, the Skopje Faculty became the fully authorised member of EAEVE. Experiences with the Slovenian model of the organisation of the Veterinary Faculty were also introduced in the Skopje Faculty system. First contacts with the colleagues from Macedonia were established in the eighties of the last century. Important events which set up the successful collaboration were scientific meetings in Ohrid and the first embryo transfer in Macedonia, carried out by the Slovenian experts. After the establishment of the Faculty of Veterinary Medicine in the autonomous Republic of Macedonia 16 years ago, the connections became closer and stronger. The cooperation continued with the active bilateral projects, expert exchanges, evaluation of study programmes, organisation of the veterinary institute, introduction of quality assurance, accreditation of diagnostic laboratories, etc. Especially important is the introduction of Bologna process in the study of the veterinary medicine, which demanded a lot of new knowledge, experience and skills. Professor Pogačnik is very honourable and respected expert in many fields of the veterinary science at national and international level. With his previous work and cooperation, professor Pogačnik essentially contributed not only to the development of the Faculty of Veterinary Medicine of the University St. Cyril and Method of Skopje, but also to the general development of the veterinary medicine in the Republic of Macedonia. At the event of the first nomination of doctor honoris causa in the veterinary profession in Slovenia, we sincerely congratulate professor Pogačnik for this great honour. Prof. Dr. Vojteh Cestnik, Veterinary Faculty, Ljubljana Slov Vet Res 2007; 44 (4): 135-6 SUBJECT INDEX VOLUME adenocarcinoma 103 adenokarcinom 108 Ambystoma mexicanum 5, 10 analize 18, 90, 132 analysis 11, 81 aniline compounds 81 anilinske spojine 90 animals 55 antibiotics 11 antibiotiki 18 antibodies, viral 123 antifungal agents 81 antimikotiki 90 bacterial toxins 91 bakterijski toksini 96 biopsija, tankoigelna 114 biopsy, fine-needle 109 blood 91, 123 bolezni 62, 96, 132 bolezni, parodontalne 108 bolnišnična okužba 122 cardiology 31 carps 81 cats 63 cattle 91 celična diferenciacija 10 cell differentiation 5 chemical analysis 123 chemotherapy 109 chromatography, liquid 81 čiščenje 30 citodiagnostika 114 Clostridium infections 91 complications, postoperative 115 contamination 25 control 63, 115 cross infection 115 cushing syndrome 73 cushingov sindrom 80 cytodiagnosis 109 dermatomikoze 72 dermatomycoses 63 desert climate 123 diagnosis 91, 123 diagnostika 96, 132 diastola 40 diastole 31 disease 55, 91, 123 44, 2007 diseases, periodontal 103 diseases, reservoirs 63 dogs 19, 31, 63, 73, 103, 109, 115 drug 11, 73, 81 echocardiography 31 ehokardiografija 40 encimsko vezan imunosorbentni test 132 enostranski kriptorhidizem 114 enzyme-linked immunosorbent assay 123 epidemije 132 epidemiologija 72 epidemiology 63 ethics, medical 55 etika, medicinska 62 etiologija 24, 122 etiology 19, 115 fish 81 fiziologija 40 food 11, 25, 81 gen MDR 24 gene expression 5 genes, MDR genetics 5, 19 genetika 10, 24 gensko izražanje 10 goats 123 govedo 96 hipofizni ACTH, hipersekrecija 80 horses 63 hrana 18, 30, 90 infection 63, 115 infekcija 72, 122 inspection 25 isolation 25 ivermectin 19, 24 izolacija 30 jetra 80 kardiologija 40 kemoterapija 114 kirurgija 114 Klebsiella infections 115 Klebsiella infekcije 122 klostridijske infekcije 96 komplikacije, pooperacijske 122 konji 72 kontaminacija 30 koze 132 krapi 90 136 Subject Index Volume 44, 2007 kri 96, 132 kromatografija, tekočinska 90 laboratorijski poskusi 62 laboratory experiments 55 liver 73 mačke 72 meat 11, 25 meso 18, 30 methods 63, 81, 123 metode 72, 90, 132 microbiological sensitivity tests 11 microbiology 25, 63, 115 mikrobiologija 30, 72, 122 mikrobni občutljivostni testi 18 molecular biology 19 molekularna biologija 24 nadzor 30, 72, 122 neoplasms 103, 109 neuroaminidase 91 neurotoxicity syndromes 19 nevraminidaza 96 nevrotoksični sindromi 24 nos 108 nose 103 novotvorbe 108, 114 ostanki 18, 90 ovce 132 pathology 63, 91, 103, 123 patofiziologija 80 patologija 72, 96, 108, 132 pedigree 19 peste-des-petits-ruminants 123, 132 P-glikoprotein 24 P-glycoprotein 19 physiology 31 physiopathology 73 piruvična kislina 96 pituitary ACTH hypersecretion 73 postrv 90 prekat, levi 40 prenos 72 protitelesa, virusna 132 psi 24, 40, 72, 80, 108, 114, 122 purification 25 puščave 132 pyruvic acid 91 raziskave 62 regeneracija 10 regeneration 5 regulacija, razvojna 10 regulation, developmental 5 residues 11, 81 rezervoarji okužbe 72 rhinitis 103, 108 ribe 90 rodovnik 24 rosaniline dyes 81 rosanilinska barvila 90 sheep 123 Staphylococcus aureus 25, 30 surgery 109 testis 109, 114 therapy 73 transmissions 63 tretionine 73, 80 trout 81 učinki zdravil 80 ultrasonography 109 ultrazvok 114 unilateral cryptorchidism 109 ventricle, left 31 ventricular function 31 veterinarska medicina 40 veterinary medicine 31 virologija 132 virology 123 zdravila 18, 90 zdravljenje 80 živali 62 živalski modeli 62 zoonoses 63 zoonoze 72 Slov Vet Res 2007; 44 (4): 137 AUTHOR INDEX VOLUME 44, 2007 Adamu S, see Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN.......91 Amupitan E, see Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN.......91 Bajc Z, Doganoc DZ, Šinigoj Gačnik K. Determination of malachite green and leucomalachite green in trout andcarp muscle by liquid chromatography with visible and fluorescence detection.................81 Balogun EO, see Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN.......91 Cabrera Blatter MF, see Ortemberg L, Loiza M, Martiarena B, Cabrera Blatter MF, Ghersevich MC, Castillo V............................73 Castillo V, see Ortemberg L, Loiza M, Martiarena B, Cabrera Blatter MF, Ghersevich MC, Castillo V.....73 Cestnik V. Prof. Dr. Milan Pogačnik - Doctor Honoris Causa ..............................133 Cotman M, Zabavnik J. Mutation of MDR1 gene associated with multidrug sensitivity in Australian shepherds in Slovenia .................... 19 Crossley DA, see Nemec A, Pavlica Z, Juntes P, Crossley DA..........................103 Doganoc DZ, see Bajc Z, Doganoc DZ, Šinigoj Gačnik K ....................... 81 Domanjko Petrič A, see Kobal M, Domanjko Petrič A . . 31 Esievo KAN, see Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN.......91 Gahlot AK, see Kataria AK, KatariaN, Gahlot AK . . 123 Ghersevich MC, see Ortemberg L, Loiza M, Martiarena B, Cabrera Blatter MF, Ghersevich MC, Castillo V . . . 73 Goršič M. Axolotl - a supermodel for tissue regeneration...........................5 Hudler P. The use of animals in biomedical research. . 55 Ibrahim ND, see Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN.......91 Juntes P, see Nemec A, Pavlica Z, Juntes P, Crossley DA..........................103 Kataria AK, KatariaN, Gahlot AK. Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert pf India ............... 123 KatariaN, see Kataria AK, KatariaN, Gahlot AK . . 123 Kirbiš A. Microbiological screening method for detection of aminoglycosides, p-lactames, macrolides, tetracyclines and quinolones in meat samples ..... 11 Kobal M, Domanjko Petrič A. Echocardiographic diastolic indices of the left ventricle in normal doberman pinschers and retrievers .................. 31 Kotnik T, see Plavec T, Tozon N, Kotnik T.......109 Kotnik T. Dermatophytoses in domestic animals and their zoonotic potential...................63 Loiza M, see Ortemberg L, Loiza M, Martiarena B, Cabrera Blatter MF, Ghersevich MC, Castillo V.....73 Martiarena B, see Ortemberg L, Loiza M, Martiarena B, Cabrera Blatter MF, Ghersevich MC, Castillo V.....73 Nemec A, Pavlica Z, Juntes P, Crossley DA. Advanced periodontal disease in a Yorkshire terrier with concurrent nasal cavity malignancy...........103 Nok AJ, see Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN.......91 Ortemberg L, Loiza M, Martiarena B, Cabrera Blatter MF, Ghersevich MC, Castillo V. Retinoic acid as a therapyfor cushing's disease in dogs: Evaluation of liver enzymes during treatment ................. 73 Pavlica Z, see Nemec A, Pavlica Z, Juntes P, Crossley DA..........................103 Pengov A, see Podpečan B, Pengov A, Vadnjal S . . . 25 Plavec T, Tozon N, Kotnik T. Generalized symmetric alopecia and hyperoestrogenism associated with concurrent lymphoma, sertoli cell tumour and seminoma in a Samoyed ..........................109 Podpečan B, Pengov A, Vadnjal S. The source of contamination of ground meat for production of meat products with bacteria Staphylococcus aureus.....25 Seliškar A, Zdovc I, Zorko B. Nosocomial Klebsiella oxytoca infection in two dogs................115 Šinigoj Gačnik K, see Bajc Z, Doganoc DZ, Šinigoj Gačnik K............................81 Tozon N, see Plavec T, Tozon N, Kotnik T........109 Useh NM, Amupitan E, Balogun EO, Adamu S, Ibrahim ND, Nok AJ, Esievo KAN. Plasma pyruvic acid changesin zebu cattle experimentally infected with Clostridium chauvoei .................... 91 Useh NM. Constraints of blackleg control in Nigeria . 101 Vadnjal S, see Podpečan B, Pengov A, Vadnjal S . . . 25 Zabavnik J, see Cotman M, Zabavnik J ........ 19 Zdovc I, see Seliškar A, Zdovc I, Zorko B.......115 Zorko B, see Seliškar A, Zdovc I, Zorko B.......115 INSTRUCTIONS FOR AUTHORS NAVODILA AVTORJEM Slovenian Veterinary Research contains original articles which have not been published or considered for publication elsewhere. All statements in the articles are the responsibility of the authors. The editorial policy is to publish original research papers, review articles, case reports and abstracts of theses, as well as other items such as critical reviews of articles published in Slov Vet Res, shorter scientific contributions, letters to the editor, etc. Authors should send their contributions to the editorial board's address. All articles are subjected to both editorial review and review by an independent referees selected by the editorial board. The editorial board reserves the right to translate titles, summaries and keywords that have not been translated into Slovene by the authors. Contributions should be written in English and should not exceed 12 pages (27 lines per page, approx. 75 characters per line). They should be submitted electronically (preferably to E-mail address, slovetres@vf.uni-lj.si), written in any word processor for Windows. Authors are requested to provide names of three potential reviewers. 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All references cited in the text should appear in the References. They should be numbered in the text in the order in which they appear, marked with Arabic numerals placed in parenthesis. The first reference in the text should determine the number and order of the respective source in the References. If the author refers again to a source which has already been used in the text, he should cite the number the source had when it was referred to for the first time. Only works which have been published or are available to the public in any other way may be referred to. Unpublished data, unpublished lectures, personal communications and similar should be mentioned in the references or footnotes at the end of the page on which they appear. Sources in the References should be listed in the order in which they appear in the text. If the source referred to was written by six authors or less, all of them should be cited; in the case of seven or more authors, only the first three should be cited, followed by 'et al.'. Any errata should be submitted to the editor-in-chief in good time after publication so that they may be publishedin the next issue. Examples of references Book: Hawkins JD. Gene structure and expression. Cambridge: University Press, 1991: 16. Chapterorar ticle in a book: Baldessarini RJ. Dopamine receptors and clinical medicine. In: Neve KA, Neve RL, eds. The dopamine receptors. Totowa: Human Press, 1996: 475-98. Article in a journal or newspaper: Fuji J, Otsu K, Zorzato F, et al. Identification of mutation in porcine ryanodine receptor asociated with malignant hyperthermia. Science 1991; 253: 448-51. Article in proceedings of a meeting or symposium: Schnoebelen CS, Louveau I, Bonneau M. Developmental pattern of GH receptor in pig skeletal muscle. In: the 6th Zavrnik memorial meeting. Lipica: Veterinary Faculty 1995: 83-6. Slovenski veterinarski zbornik (Slovenian Veterinary Research) objavlja izvirne prispevke, ki še niso bili objavljeni oz. poslani v objavo drugam. Za vse navedbe v prispevkih so odgovorni avtorji. Uredniška politika obsega publiciranje znanstvenih člankov, preglednih znanstvenih člankov, strokovnih člankov, povzetkov disertacij in drugih prispevkov, kot so kritične presoje o vsebini razprav, objavljenih v zborniku, kratke znanstvene prispevke, pisma uredniku in drugo. Avtorji pošljejo prispevke na naslov uredništva. Glavni urednik pregleda vse prispevke. Za vse članke je obvezna strokovna recenzija, za katero poskrbi uredništvo. Prispevki naj bodo napisani v angleškem jeziku, z naslovom, povzetkom in ključnimi besedami tudi v slovenščini. Obsegajo naj največ 12 strani, kar pomeni 27 vrstic na stran s približno 75 znaki v vrstici. Prispevki naj bodo poslani v elektronski obliki v katerem koli urejevalniku besedil za okensko okolje. Zaželjena je uporaba elektronske pošte (slove-tres@vf.uni-lj.si) in avtorji naj predlagajo tri možne recenzente. Besedilo naj ima dvojni razmik med vrsticami, pri čemer naj bodo vrstice na levi strani oštevilčene. Besedilo naj bo na levi strani od roba oddaljeno 4 cm. Naslovna stran prispevkov se začne z naslovom, sledi ime in priimek avtorja. Kadar je avtorjev več, jih ločimo z vejicami. V naslednjih vrsticah je v rubriki Addresses of authors: za dvopičjem treba navesti polno ime in priimek ter naslov(e) avtorja(ev), tj. ustanovo, ulico s hišno številko, pošto in kraj. Vse navedene podatke ločujejo vejice. Sledi vrstica, kjer je treba navesti ime ter elektronski (E-mail:) in poštni naslov ter telefonsko številko (Phone:) odgovornega avtorja. Sledi besedilo povzetka Summary v obsegu 16 do 20 vrstic (približno 1000 do 1500 znakov). V naslednji rubriki Key words: se za dvopičjem navedejo ključne besede. Posamezne besede ali sklopi besed morajo biti ločeni s podpičjem. Znanstveni članki in tisti, ki so prikaz lastnih raziskav in dognanj, morajo vsebovati še naslednje obvezne rubrike, s katerimi avtor sam naslovi ustrezne dele besedila v prispevku: Introduction, Material and methods, Results, Discussion in References. Pregledni članki naj vsebujejo uvod, poglavja, ki so glede na vsebino smiselno naslovljena, in literaturo. Podatke o financerjih ali drugih zadevah, pomembnih za prispevek, npr. o tehnični pomoči, avtorji navedejo v rubriki Acknowledgements, ki se uvrsti pred rubriko References. Za rubriko References sledijo spremna besedila k slikam. Priloge, kot so tabele, grafikoni in diagrami naj bodo smiselno vključene v besedilo. Slikovni material naj bo poslan posebej v obliki bmp, jpg, ali tif. Priloge in slike morajo biti poimenovane z besedami, ki jih opredeljujejo, in arabskimi številkami (npr. Table 1:, Figure 1: itn.). Za dvopičjem sledi besedilo oziroma naslov. Vsi navedki (reference), citirani v besedilu, se morajo nanašati na seznam literature. V besedilu jih je treba oštevilčiti po vrstnem redu, po katerem se pojavljajo, z arabskimi številkami v oklepaju. Prvi navedek v besedilu opredeli številko oziroma vrstni red ustreznega vira v seznamu literature. Če se avtor v besedilu ponovno sklicuje na že uporabljeni vir, navede tisto številko, ki jo je vir dobil pri prvem navedku. Citirana so lahko le dela, ki so tiskana ali kako drugače razmnožena in dostopna javnosti. Neobjavljeni podatki, neobjavljena predavanja, osebna sporočila in podobno naj bodo omenjeni v navedkih ali opombah na koncu tiste strani, kjer so navedeni. V seznamu literature so viri urejeni po vrstnem redu. Če je citirani vir napisalo šest ali manj avtorjev, je treba navesti vse; pri sedmih ali več avtorjih se navedejo prvi trije in doda et al. Da bi se morebitni popravki lahko objavili v naslednji številki, jih morajo avtorji pravočasno sporočiti glavnemu uredniku. Načini citiranja Knjiga: Hawkins JD. Gene structure and expression. Cambridge: University Press, 1991: 16. Poglavje ali prispevek v knjigi: Baldessarini RJ. Dopamine receptors and clinical medicine. In: Neve KA, Neve RL, eds. The dopamine receptors. Totowa: Human Press, 1996: 475-98. Članek iz revije ali časopisa: Fuji J, Otsu K, Zorzato F, et al. Identification of mutation in porcine ryanodine receptor asociated with malignant hyperthermia. Science 1991; 253: 448-51. Članek iz zbornika referatov: Schnoebelen CS, Louveau I, Bonneau M. Developmental pattern of GH receptor in pig skeletal muscle. In: the 6th Zavrnik memorial meeting. Lipica: Veterinary Faculty 1995: 83-6. Slov Vet Res 2007; 44 (4) Letter to Editor Useh NM. Constraints of blackleg control in Nigeria Case Reports Nemec A, Pavlica Z, Juntes P, Crossley DA. Advanced periodontal disease in a Yorkshire terrier with concurrent nasal cavity malignancy.................................................................103 Plavec T, Tozon N, Kotnik T. Generalized symmetric alopecia and hyperoestrogenism associated with concurrent lymphoma, sertoli cell tumour and seminoma in a Samoyed.......................................109 Seliskar A, Zdovc I, Zorko B. Nosocomial Klebsiella oxytoca infection in two dogs.........................115 Kataria AK, Kataria N, Gahlot AK. Large scale outbreaks of peste des petits ruminants in sheep and goats in Thar desert pf India....................................................................123 Cestnik V. Prof. dr. Milan Pogacnik - Doctor honoris causa..........................................133 Subject Index Volume 44, 2007............................................................... 135 Author Index Volume 44, 2007 ............................................................... 137