For many years, the conflict between humans and wolves has persisted due to the death of livestock attributed to the attack of these animals and dogs, causing high economic costs to owners and ...governments. To remedy this problem, differential compensation programs have been established for the affected owners, depending on the attacker. Obtaining these benefits requires evidence to demonstrate the veracity of the complaint. Reliable approaches are needed nowadays to detect the predator, beyond any reasonable doubt. Although the analysis of teeth marks on bones has been used to differentiate carnivores, especially for archaeological purposes, its interpretation for forensic purposes is still ambiguous, due to the high range of factors that can influence tooth mark patterns, and it has scarcely been considered in previous work. This study analyzed and compared the tooth marks caused by captive Iberian wolves and by a group of domestic hunting dogs (rehala) on fresh, and disarticulated (isolated) bovine scapulae, for taxonomic differentiation purposes. Wolves showed a higher rate of modification throughout the study and, although tooth marks caused by wolves tend to be larger and wider than those caused by dogs, in most cases it was possible to find overlap between the two subspecies. Bone modifications are conditioned by a number of factors intrinsic to the scavenger or predator species, and intrinsic to the aggressor and the environment, that must be considered during the interpretation of tooth marks found on bones at a crime scene. Along with the comprehensive analysis of all evidence, the analysis of new variables of tooth marks on bones, using novel image processing methodologies and statistical analysis, has shown high potential to identify the morphological and/or morphometric variables that allow taxonomic differentiation
•Iberian wolf and domestic dog can be differentiated by their bone bite mark patterns.•The forensic analysis of bite marks of both canids must be interdisciplinary.•The bite marks patterns of both canids have a multifactorial origin.
One of the current leading causes of death in pet cats is neoplasia. The scarcity of data on tumor prevalence in cats from south-eastern Europe led to this retrospective study which reports an ...abundant collection of tumors diagnosed in cats living in Croatia. Archived histopathology findings (diagnosis, cell origin, and biological behavior), and epidemiological data (breed, sex, age, and anatomical site of the tumor) of the cats diagnosed with at least one tumor were collected and statistically analyzed. For the research, 2338 archived feline samples, obtained from 2009-2019, have been reviewed. In 659 samples (28.1%) one or more tumors were diagnosed. Tumors have most often affected non-pedigree cats, cats between 7 and 15 years of age, and female cats. Most tumors showed malignant behavior (85.7%). The most frequent diagnoses were mammary adenocarcinoma, lymphoma, and squamous cell carcinoma. Regarding origin, most samples were epithelial (46.9%), but mesenchymal (28.7%) and lymphoid tumors (17.0%) were also frequently found. Tumors most often occurred in the skin and subcutis, mammary gland (in females), digestive system, and hemolymphatic system (in males). Statistical analysis did not reveal breed, sex, and age predisposition, but female animals and non-pedigree cats were significantly more likely to develop malignant tumors. This detailed study gives insight into the most frequent tumors that occur in cats from Croatia, and the data reported here is likely applicable to other countries. This data will contribute to clinical veterinary oncology, as it presents the most extensive tumor index of cats in south-eastern Europe.
Circumanal gland (CG) tumors are common neoplasms of older dogs. Of these, only circumanal gland carcinomas show clear metastatic potential, and even so metastases seem to be uncommon. ...Epithelial-mesenchymal transition (EMT) is a process that facilitates the initial steps of metastasis, and to date it has not been systematically investigated in CG tumors. Insight into the occurrence of this process would be a valuable asset in understanding the biology of these tumors. To test the occurrence of EMT we used three immunohistochemical markers that alter their expression in this process, namely – E-cadherin, N-cadherin, and matrix metalloproteinase-9 (MMP-9). Additionally, we used the Ki-67 marker of proliferation. The samples used consisted of 15 adenomas, 11 epitheliomas, 21 well-differentiated carcinomas, seven poorly differentiated carcinomas, and ten samples of normal CG. The results of N-cadherin were negative for all samples. E-cadherin was highly expressed in all groups, but was slightly lower in semi-malignant and malignant tumors; MMP-9 marking was generally very low, but significantly higher in semi-malignant or malignant tumors when compared to benign or non-neoplastic CG. The index of proliferation (Ki-67) was significantly higher for semi-malignant or malignant CG tumors when compared to benign CG tumors or normal CG. These results show that with an increase in the histologic malignancy of these tumors there is a slight drop in E-cadherin, a slight rise in MMP-9, and a significant increase in Ki-67. Therefore, these results suggest the possibility that EMT occurs within the malignant or even semi-malignant forms of CG tumors, but probably as a rare and late event. Further studies are needed to prove or disprove these statements.
Biogenic amines (BAs) are considered a potential microbiological toxicological hazard in aged cheese. Risk mitigation strategies include good hygiene practice measures, thermal treatment of milk and ...the use of competitive dairy cultures. The aim of this study was to evaluate the amount of BAs—tryptamine, β-phenylethylamine, putrescine, cadaverine, histamine, tyramine, spermidine and spermine—in the core and rind of cheeses ripened by bacteria (n = 61) and by mold cultures (n = 8). The microbial communities were counted, and the dominant lactic acid bacteria (LAB) were identified, corresponding to the BA concentrations. The total BA content was highest in the core of semi-hard cheeses (353.98 mg/kg), followed by mold cheeses (248.99 mg/kg) and lowest in hard cheeses (157.38 mg/kg). The highest amount of BAs was present in the rind of cheeses with mold (240.52 mg/kg), followed by semi-hard (174.99 mg/kg) and hard cheeses (107.21 mg/kg). Tyramine was the most abundant BA, represented by 75.4% in mold cheeses, 41.3% in hard cheese and 35% of total BAs in semi-hard cheeses. Histamine was present above the defined European maximum level (ML) of 100 mg/kg in only two semi-hard and three hard cheeses. High amount of BAs (above 600 mg/kg) in cheeses, mainly tyramine, were associated with the presence of Enterococcus durans, while negligible BA concentrations were found in cheeses ripened with Lacticaseibacillus rhamnosus, Lactococcus lactis or Lacticaseibacillus paracasei cultures. This study has shown that retailed cheese varieties produced with commercial bacterial or mold cultures have acceptable levels of biogenic amines with respect to consumers.
SEVERIN, K, P. DZAJA, M. SPERANDA, M. DIDARA, D. KONJEVIC, E. SATROVIC, K STARCEVIC: Estimation of red deer (Cervus elaphus) post mortal interval based on the biochemical parameters of vitreous fluid ...using linear regression analyses. Vet. arhiv 88, 511-519, 2018. Vitreous fluid is a stable biological fluid, which is well-protected from contamination and post mortem degradation, which makes it suitable in forensic pathology for determination of the post mortem interval (PMI). The present study was conducted to discover the correlation between increasing PMI and levels of various vitreous biochemical parameters (sodium, calcium, magnesium, chloride, potassium, phosphor, AST, urea, creatinine, total protein, glucose, total cholesterol). The estimation of PMI in red deer is of great interest in cases of illegal hunting. The vitreous humour samples were collected from six red deer carcasses. The eyes were enucleated, covered by parafilm and aluminium foil, packed in a small container and maintained at +4 degreesC. Sampling of vitreous fluid was done again at 8, 10 and then regularly each 10 hours until finally 90 hours after death. A significant linear correlation was found for potassium, sodium and chloride (P<0.0001). The present study revealed that biochemical analysis of vitreous fluid could be used to determine post mortem interval in red deer. Key words: vitreous fluid; potassium; sodium; chloride; red deer
Visceral hemangiosarcoma were analyzed at the Department of Veterinary Pathology of the Faculty of Veterinary Medicine, University of Zagreb, over a 5-year period. From a total of 52 tumor masses in ...a total of 31 dogs, histological growth patterns (cavernous, capillary or solid) and the amount of tumor supporting stroma were evaluated. Additionally, sections were stained with toluidine blue for the purpose of mast cell detection and their number was determined in the tumor parenchyma, tumor stroma and edges. The average age of the affected animals was 10 years, males predominated, and the tumors occurred most frequently in cross breeds, German Shepherd dogs and Labrador Retrievers. The highest number of visceral hemangiosarcoma was found in the spleen (27/52). The most common growth pattern of visceral hemangiosarcoma was solely cavernous with a mixture of cavernous and solid patterns in different proportions. Mast cells were found in 44/52 (84.6%) of the visceral hemangiosarcomas. A smaller number of mast cells were found in the tumor parenchyma, and higher number were found in the stroma and tumor edges. The number of mast cells in tumors was not significantly associated with the tumor growth pattern, but there was a positive correlation between MCC - tumor parenchyma and stroma (rs = 0.28, P<0.05), MCC - tumor stroma + edges (rs = 0.74, P<0.05) and MCC - tumor parenchyma and MCC- tumor stroma+edges (rs = 0.30, P<0.05) in all the examined tumors. In splenic tumors, there was only a significant positive correlation between MCC - stroma+edges (rs P = 0.68 P<0.05). These results suggest a higher mast cell count in tumors with more developed stromal components in canine visceral hemangiosarcomas, and certainly indicate the need for further research on their role and the factors they release in the development and progression of hemangiosarcomas.
Testicular tumors are the most common genital neoplasms in male dogs, with Leydig cell tumors (LCT), seminomas (SEM), and Sertoli cell tumors (SCT) the most common forms. Human SEM are classified as ...classical (CSEM) or spermatocytic (SSEM). Intratubular germ cell neoplasia of undifferentiated origin (IGCNU) is another form of human testicular tumor. The aim of this study was to verify that CSEM/SSEM classification is valid in dogs and confirm the existence of canine IGCNU.
Testicular tumors were found in 46% of dogs at necropsy and accounted for 7% of tumors biopsied. The median age of dogs with tumors at necropsy was 10.16 years; median age at positive biopsy was 10.24 years. The most common tumors, in decreasing order, were LCT, mixed tumors, SEM and SCT at necropsy, and SEM, SCT, mixed tumors, LCT, peripheral nerve sheath tumor, and teratoma in the biopsy group. IGCNU was found in 3% of testicles at necropsy and in 3% of biopsy samples. Two dogs had testicular tumor metastasis. Expression of c-KIT was most common in SEM and seminomatous components of mixed tumors. PLAP was mostly expressed in IGCNU, SEM, teratoma, and some mixed tumors. Cytokeratin was mainly expressed in SCT. CD30 expression was low in both groups.
The high tumor incidence at necropsy can be attributed to older age. Tumor incidence in biopsy samples, dog age, and histological classification were consistent with previous studies. The higher incidence of SEM and SCT in the biopsy group probably resulted from the obvious clinical expression of these tumor types. The low incidence of metastasis confirmed the predominance of benign tumors. Low CD30 expression confirmed the low incidence of testicular embryonal carcinoma. Cytokeratin helps differentiate stromal tumors, especially SCT, from germ cell tumors. Histology and c-KIT and PLAP expression indicate that IGCNU exists in dogs. Expression of c-KIT and PLAP confirmed that CSEM and SSEM classification is valid in dogs.
Measurement of faecal glucocorticoid metabolites is increasingly used as a non-invasive tool to examine disturbances in various domestic and wild animals. Because measurements of faecal ...glucocorticoid metabolites has previously never been reported in fallow deer, we determined 11,17-dioxoandrostanes (11,17-DOA), a group of cortisol metabolites, in the faeces of four fallow deer yearlings after an adrenocorticotropic hormone (ACTH) challenge or control saline injection by an 11-oxoaetiocholanolone enzyme immunoassay (EIA), to validate a method. A 2.9- to 4.3-fold increase in measured cortisol metabolites in challenged animals after approximately 22 h demonstrated the suitability of this group-specific EIA to monitor adrenocortical activity in respective deer species. To determine faecal cortisol metabolites in fallow deer from a Mediterranean habitat, we collected samples during a 1-year study at Veliki Brijuni Island. The study confirmed seasonal pattern of cortisol release in fallow deer. Higher 11,17-DOA concentrations (median; min-max) were determined for November (99; 50-2,035), March (112; 25-315) and May (92; 40-196 ng/g faeces). Significantly lower concentrations were measured during July (30; 10-195 ng/g faeces). This study indicates that the analysis of faecal glucocorticoid metabolites is a valuable non-invasive technique for monitoring adrenocortical activity in fallow deer. This, together with information about the seasonal pattern of glucocorticoid excretion, could help to improve fallow deer management and welfare, especially in the case of farmed and park animals.
The Postmortem Interval (PMI) is the time from the death of an animal to its discovery. From a veterinary forensic standpoint, an accurate estimation of the PMI is of particular importance, ...especially with the observed increase in deaths of domestic and wild animals. A preliminary study was conducted using the eyes of domestic pigs. A biochemical analysis was conducted on the vitreous humor of the eye, whilst a histological analysis was conducted on the retina. The eyes were stored at +4 °C and changes were assessed at time intervals of 0, 12, 24, 48, and 120 h. The biochemical analysis during the PMI established a decrease in sodium, chlorine, and glucose concentrations, and a rise in potassium concentration. Accordingly, a simple linear regression showed a significant correlation between changes in concentrations of sodium (Na
), potassium (K
), chloride (Cl
), and glucose, in relation to the PMI. The histological analysis showed evident morphological changes in the retina, which included homogenization of the rod and cone cells, pyknosis of the outer nuclear layer, homogenization of the outer plexiform layer, pyknosis of the inner nuclear layer, homogenization of the inner plexiform layer, and pyknosis of the nuclei of the ganglion layer of the retina.
The aim of this research was to investigate the use of an indirect enzyme-linked immunosorbent assay (ELISA) and Western blotting test with native excretory/secretory Fascioloides magna antigen ...(ES-Ag) in free-ranging red deer (Cervus elaphus). The research was performed on 48 red deer shot during routine culls from two different areas, considering the occurrence of fascioloidosis. After coprological examination and gross pathology examination of the livers for F. magna, serum samples were divided into three groups as infected (n = 32), uninfected (n = 13) and previously infected deer (n = 3). Indirect ELISA results were significantly higher for serum samples from infected deer (percentage of positivity (PP) 65.1 ± 18.4) than uninfected (PP 11.6 ± 13.7) and previously infected deer (PP 20.3 + or - 6.4). Samples from uninfected deer had fewer bands (30-33 and 104 kDa) on Western blotting than samples from infected deer (6, 17, 22, 27, 30-33, 40, 45 and 104 kDa). The number of alive flukes positively correlated with the ELISA results and the number of F. magna eggs per gram of faeces (EPG). The results indicated that ELISA and Western blotting test could be useful in the early diagnosis of fascioloidosis in red deer and also in control in the population of free-ranging red deer. Key words: Fascioloides magna, red deer, excretory/secretory antigens, indirect ELISA, Western blotting Cilj ovog rada bio je istraziti imunodijagnosticki potencijal ekskretomo/sekretomog antigena (ES-Ag) dobivenog od odraslih Fascioloides magna u populaciji slobodno zivucih jelena obicnih primjenom neizravnoga imunoenzimnog i Western blotting testa. Istrazivanje je provedeno na 48 jelena obicnih odstrijeljenih u okviru planiranog odstrjela na dva razlicita podrucja s obzirom na pojavu fascioloidoze. Nakon koproloske pretrage i razudbe jetara na prisutnost F. magna, uzorci seruma bili su podijeljeni u tri skupine: invadirani (n = 32), neinvadirani (n = 13) i prethodno invadirani jeleni (n = 3). Rezultati neizravnog ELISA testa bili su znacajno visi u uzorcima seruma od invadiranih jelena (postotak pozitivnosti (PP) 65,08 ± 18,40) nego neinvadiranih (PP 11,6 ± 13,7) i prethodno invadiranih jelena (PP 20,31 ± 6,43). Uzorci od invadiranih imali su nekoliko linija (30-33 i 104 kDa) primjenom Western blottinga za razliku od uzoraka od invadiranih jelena (6, 17, 22, 27, 3033, 40, 45 i 104 kDa). Broj zivih metilja u pozitivnoj je korelaciji s rezultatima ELISA testa i broja F. magna jajasca po gramu izmeta (EPG). Rezultati ukazuju da neizravna ELISA i Western blotting test mogu biti korisni s ciljem ranog otkrivanja fascioloidoze u jelena obicnog te isto tako u njenoj kontroli u populaciji slobodno zivucih jelena obicnih. Kljucne rijeci: Fascioloides magna, jelen obicni, ekskretorno/sekretorni antigeni, neizravna ELISA, Western blotting
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