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Antlers are bony structures that undergo regular annual growth, mineralisation and casting phases, representing only mammalian organs capable of full regeneration. Myiasis is infestation of live vertebrates with dipterous larvae. We sampled mineralised antlers from a red deer spiker stag 2 months after velvet shedding, divided it into three segments and fixed in 10% neutral buffered formalin. After demineralisation, samples were embedded in paraffin and sliced to a thickness of 6 µm. Tissue sections were stained with hematoxylin–eosin (HE), a modified staining to show the ossification process, toluidin blue and Masson’s trichrome staining. Smears of liquid content from the antlers were made and stained with May-Grünwald Giemsa. Larvae were separated from segments and preserved in 70% ethanol for identification. Macroscopically, some parts of the antler tips were lacking the compact part. Microscopically, within the Haversian and Volkmann canals, a large number of bacteria, scarce protein content and remnants of red blood corpuscles were visible. In the area of cancellous bone, cross-sectioned larvae were present. A large quantity of bacteria and a few degraded red blood corpuscles were visible on the smear made of liquid from the antlers. For morphological identification, three larvae were examined: two were third-instar larvae (L3), while one was a first instar larva (L1). Based on the shape of the cephaloskeleton, L3 was identified as Prochyliza nigrimanus and confirmed using molecular tools. To the extent of the authors’ knowledge, this is the first record of Prochyliza nigrimanus in non-casted hard red deer antlers and the first description of this species in Croatia. Graphical abstract

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.

Background 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. Results 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. Conclusions 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.

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.

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.

Antlers are bony structures that undergo regular annual growth, mineralisation and casting phases, representing only mammalian organs capable of full regeneration. Myiasis is infestation of live vertebrates with dipterous larvae. We sampled mineralised antlers from a red deer spiker stag 2 months after velvet shedding, divided it into three segments and fixed in 10% neutral buffered formalin. After demineralisation, samples were embedded in paraffin and sliced to a thickness of 6 ?. Tissue sections were stained with hematoxylin-eosin (HE), a modified staining to show the ossification process, toluidin blue and Masson's trichrome staining. Smears of liquid content from the antlers were made and stained with May-Gr?nwald Giemsa. Larvae were separated from segments and preserved in 70% ethanol for identification. Macroscopically, some parts of the antler tips were lacking the compact part. Microscopically, within the Haversian and Volkmann canals, a large number of bacteria, scarce protein content and remnants of red blood corpuscles were visible. In the area of cancellous bone, cross-sectioned larvae were present. A large quantity of bacteria and a few degraded red blood corpuscles were visible on the smear made of liquid from the antlers. For morphological identification, three larvae were examined: two were third-instar larvae (L3), while one was a first instar larva (L1). Based on the shape of the cephaloskeleton, L3 was identified as Prochyliza nigrimanus and confirmed using molecular tools. To the extent of the authors' knowledge, this is the first record of Prochyliza nigrimanus in non-casted hard red deer antlers and the first description of this species in Croatia. Graphical abstract

During the armed conflict in Colombia, homemade improvised antipersonnel landmines were used to neutralize the adversary. Many active artifacts remain buried, causing damage to biodiversity by exploding. The extensive literature describes the effects and injuries caused to humans by conventional landmines. However, there is considerably less information on the behavior and effects of homemade antipersonnel landmines on fauna and good field investigation practices. Our objectives were to describe the characteristics of a controlled explosion of a homemade antipersonnel landmine (using ammonium nitrate as an explosive substance), to compare the effectiveness of some evidence search patterns used in forensic investigation, and to determine the effects on a piece of an animal carcass. The explosion generated a shock wave and an exothermic reaction, generating physical effects on the ground and surrounding structures near the point of explosion. The amputation of the foot in direct contact with the device during the explosion and multiple fractures were the main effects on the animal carcass. Finally, it was determined that finding evidence was more effective in a smaller search area. Many factors can influence the results, which must be weighed when interpreting the results, as discussed in this manuscript.

The aim of the present study was to determine the reference intervals for the most commonly used hematological and biochemical parameters of European mouflon from a closed hunting ground in the eastern part of the Republic of Croatia. Blood samples were collected from 39 live, physically restrained, clinically normal European mouflon, as well as from 50 domestic sheep. The distribution of values within each parameter was determined and statistical differences in values between sexes were also determined. For each sample, 14 hematological and 18 biochemical parameters were analyzed. Hematology and biochemistry values of the European mouflon were also compared with the values of domestic sheep. In further studies, the established values might be useful for the health assessment of mouflon.

During November 2006, two live and one dead mouflon (Ovis ammon musimon) were presented with a history of weakness, tremors, and paralysis. After a detailed gross and histologic examination and a bacteriologic, parasitologic, and rabies evaluation, a preliminary diagnosis of tick paralysis was established. A thorough field search revealed 13 affected mouflons found in the open hunting ground “Sveti Juraj” near the town of Senj (Croatia), along with an additional 35 mouflon carcasses. All 13 mouflons were placed in a quiet, semidark stable. All detectable ticks were removed manually, and the animals were topically treated with 250 ppm of Amitraz water emulsion (Taktic 12.5% EC, Intervet International, 5830 Boxmeer, Netherlands). The collected ticks were identified as Ixodes ricinus, Dermacentor marginatus, and Haemaphysalis punctata. In the following 24 hr, all treated animals recovered fully. This report describes a naturally occurring outbreak of tick paralysis in free-ranging mouflons from a karst habitat.