Milijan Jovanović

Survey of Slaughtered Pigs for Occurrence of Ochratoxin A and Porcine Nephropathy in Serbia

Samples of blood, kidney and liver were randomly selected from slaughtered pigs (n=90) and analyzed for ochratoxin A by HPLC. In addition, in order to obtain information on the occurrence of nephropathy, histological examinations were carried out. Of the 90 liver samples, 26.6% contained OTA in the range of 0.22–14.5 ng/g. The incidence of OTA in serum and kidney were very similar (31%, 33.3%), with a maximum concentration of 220.8 ng/mL, and 52.5 ng/g, respectively. Histopathological examination of kidneys confirmed tubulopathies with edema and cell vacuolization. In addition, hemorrhages and necrosis of proximal kidney tubules’ cells were found.

Thyroid Hormones Concentrations during the Mid-Dry Period: An Early Indicator of Fatty Liver in Holstein-Friesian Dairy Cows

Relationship between postpartal fatty liver and thyroid gland activity during the peripartal and mid dry periods was studied. Twenty one dry cows were chosen. Blood samples were obtained on days −30, −2, and +12 related to calving and analized for thyroxine (T4) and triiodothyronine (T3). A T3/T4 ratio was calculated. Liver tissue samples were taken 12 d after calving and tested for the lipid content. Cows were divided into three groups: mild (<20% fat), moderate (20 to 30%), or severe fatty liver (>30%). Cows, that were affected with severe fatty liver, were hypothyroid prior to development of the condition due to lower T4 concentrations, and had significantly lower concentration of T3 and higher T3/T4 ratios than cows with mild and moderate fatty liver. Thus, hypothyroid state during mid-dry period may be an early indicator of postpartal fatty liver and may provoke T3/T4 ratio increase in this group of cows.

The role of hepatic myofibroblasts in liver cirrhosis in fallow deer (Dama dama) naturally infected with giant liver fluke (Fascioloides magna).

BackgroundThis paper describes liver cirrhosis in 35 fallow deer infected with the giant liver fluke, as well as the distribution, origin, and role of myofibroblasts in its development.ResultsIn liver of infected deer, stripes of connective tissue are wound around groups of degenerated and regenerated liver lobuli. In the connective tissue, lymphocytes and macrophages which often contain parasite hematin are also present. The walls of the bile ducts are thickened, the epithelium multiplied with mucous metaplasia, and desquamated cells, parasite eggs and brown pigment are present in their lumen.In the livers with cirrhosis, immunopositivity to α-SMA and desmin was observed in cells in portal and septal spaces, at the edge between fibrotic septa and the surrounding parenchyma and in perisinusoidal spaces. These cells vary in size, they are round, oval, spindle-shaped or irregular in shape, similar to vascular smooth muscle cells. The derangement of epithelial-mesenchymal interactions detected in chronic cholangiopathies is most probably the pro-fibrogenic mechanism in liver cirrhosis of fallow deer (Dama dama) infected with the giant liver fluke (Fascioloides magna).ConclusionMyofibroblasts, especially hepatic stellate cells (HSCs), play an important role in the synthesis of extracellular matrix components in the development of parasitic fibrosis and cirrhosis in the liver of fallow deer.

NEW INSIGHTS INTO BODY CONDITION SCORE AND ITS ASSOCIATION WITH FATTY LIVER IN HOLSTEIN DAIRY COWS

The aim of this study was to estimate the association between body condition and fatty liver in high-yielding dairy cows. One hundred dry Holstein cows were selected. Cows were scored once for body condition during the dry period, puerperium and month 2 of lactation, according to the system provided by Elanco Animal Health Buletin Al 8478. Body condition loss was determined as marked if loss was over 0.7 points between two consecutive phases of cycles and over 1.5 points between puerperium and month 2 of lactation. Liver tissue samples were taken 12 d after calving and tested for lipid content. 50% of cows had mild (3.92 ± 3.33% fat), 33% moderate (19.28 ± 5.18% fat), and 17% severe fatty liver (36.21 ± 4.55% fat). The mean body condition scores were 3.79 ± 0.55 (dry period), 3.18 ± 0.34 (puerperium) and 2.90 ± 0.29 (month 2 of lactation). Marked body condition loss from the dry period to puerperium had 38% of cows, 20% from dry period to month 2 of lactation and 8% from puerperium to month 2 of lactation. Body condition scores out of the physiological range had 39% dry cows, 49 % cows in puerperium and 9% cows at month 2 of lactation. After dividing cows into groups according to fatty liver degree, it was shown that only cows with severe fatty liver were obese during the dry period and that 76.47% of cows from this group had marked condition loss from the dry period to puerperium, 47.06% from dry period to month 2 of lactation and 23.53% from puerperium to month 2 of lactation. These indicate that in cows with severe fatty liver lipomobilisation is intensive and starts around calving, before milk production enhacement. Our results indicate a strong association between body condition and fatty liver in cows, which can be estimated only when body condition is analyzed in details. Besides, our results clearly indicate that body condition is not the only etiological factor that leads to fatty liver in dairy cows.

Toxicological Assessment of Toxic Element Residues in Swine Kidney and Its Role in Public Health Risk Assessment

In order to ensure the safety of consumers in Serbia the prevalence of toxic elements (As, Cd, Hg, Pb) in swine kidney collected from three different areas in Serbia (n = 90) was determined by atomic absorption spectrometry. Also, in order to find information on the effects of accumulation of toxic elements on swine kidney, pathohistological examination of the kidneys was performed. The presence of mercury was found in 33.3% of kidney samples in the range of 0.005–0.055 mg/kg, while the presence of cadmium was detected less often (27.7%) but in larger amounts (0.05–1.23 mg/kg). The presence of arsenic was found only in one sample, while no lead was found. The results of the metal-to-metal correlation analysis supported there were the result of different sources of contamination. Pathohistological examination of kidneys confirms tubulopathies with oedema and cell vacuolization. In addition, haemorrhages and necrosis of proximal kidney tubule cells were found. This study demonstrates that toxic elements in Serbian slaughtered pigs are found at levels comparable to those reported in other countries, and consequently the levels reported in this study do not represent a concern from a consumer safety point of view. The lack of a strong correlation between histopathological changes and the incidence of toxic elements found in this study might be explained as the result of synergism among toxic elements and other nephrotoxic compounds which enhance the toxicity of the individual toxins even at the relatively low mean concentrations observed in this study.

An outbreak of fumonisin toxicosis in horses in Serbia

This paper reports on an outbreak of fumonisin toxicosis in a stable with 100 horses. Twenty-one horses were affected and fifteen died within a month. The animals were not febrile and exhibited poor appetite, somnolence, head pressing, blindness, ataxia, lethargy, and seizures, ending with death. The post mortem examination of the head of one 18-month-old colt revealed large, bilateral, slightly asymmetrical large areas of necrosis of the cerebral white matter. The lesions involved the frontal, parietal and occipital lobes only. Histological findings revealed leukomalacia with necrosis, infiltration of macrophages, neutrophils and eosinophils, haemorrhage and oedema of the surrounding grey matter. In response to the pathological findings, which indicated equine leukoencephalomalacia (ELEM), feed samples from the premises were tested for fumonisin B1 and B2 (FB1 and FB2). The milled maize samples contained 6.0 mg/kg FB1 and 2.4 mg/kg FB2, while the maize bran contained 6.05 mg/kg FB1 and 1.68 mg/kg FB2. Th...

Role of hepatic stellate cells (HSCs) in the development of hepatic fibrosis in cats with polycystic kidney disease (PKD).

Hepatic stellate cells (HSCs) play a significant role in hepatic fibrogenesis. In the following study we described the distribution of cells that express alpha-smooth muscle actin (α-SMA) and desmin in the cat liver with various degrees of fibrosis, as well as the significance of hepatic stellate cells and portal myofibroblasts in the genesis of fibrosis in cats with polycistic kidney disease. Liver samples from 15 necropsied Persian cats were examined microscopically, using H&E and Masson-trichrom methods and immunohistology for α-SMA and desmin. Liver fibrosis was confirmed in cats with terminal stage of chronic cholangiohepatitis and it was characterized by connective tissue septa which divide the liver parenchyma into irregular lobuli. Inflammation in the cat liver is connected with the activation of periductal myofibroblasts. The intensity of immunopositivity of perisinusoidal HSCs to α-SMA and desmin varied depending on the degree of fibrosis and was the strongest in livers of cats with cirrhosis.

A Survey of Spontaneous Occurrence of Ochratoxin A Residues in Chicken Tissues and Concurrence With Histopathological Changes in Liver and Kidneys

Toxicological and histopathological investigations of tissues of commercially slaughtered chickens were carried out to provide a preliminary evaluation of the incidence of occurrence of ochratoxin A (OTA) in chicken sold in Serbian retail market. In addition, the etiology of nephropathies of these chickens was elucidated. The majority of these tissue samples were not found to contain measurable amounts of OTA. Moreover, the OTA levels found in analyzed tissues were generally low and there was no positive correlation between the presence of OTA and the frequency of histopathological changes. Histopathological changes such as degenerative changes in the kidneys and liver differed from the classical description of the mycotoxic nephropathy, indicating that the chicken nephropathy observed in Serbia may have a multitoxic etiology with possible synergistic effect between microorganisms and natural toxins, usually present in low concentrations. The low OTA results also suggested that chicken meat available in the retail market in Serbia are unlikely to pose any significant adverse health risk to the consumers with respect to OTA toxicity.

Comparative investigation of the efficacy of three different adsorbents against OTA-induced toxicity in broiler chickens.

The aim of our study was to determine the efficacy of three different adsorbents, inorganic (modified zeolite), organic (esterified glucomannans) and mixed (inorganic and organic components, with the addition of enzymes), in protecting broilers from the toxic effects of ochratoxin A in feed. Broilers were fed diets containing 2 mg/kg of ochratoxin A (OTA) and supplemented with adsorbents at the recommended concentration of 2 g/kg for 21 days. The presence of OTA led to a notable reduction in body weight, lower weight gain, increased feed conversion and induced histopathological changes in the liver and kidneys. The presence of inorganic, organic and mixed adsorbents in contaminated feed only partially reduced the negative effects of OTA on the broiler performances. Broilers that were fed with adsorbent-supplemented feed reached higher body weight (17.96%, 19.09% and 13.59%), compared to the group that received only OTA. The presence of adsorbents partially alleviated the reduction in feed consumption (22.68%, 12.91% and 10.59%), and a similar effect was observed with feed conversion. The applied adsorbents have also reduced the intensity of histopathological changes caused by OTA; however, they were not able to prevent their onset. After the withdrawal of the toxin and adsorbents from the feed (21–42 days), all previously observed disturbances in broilers were reduced, but more remarkably in broilers fed with adsorbents.

The role and immunophenotypic characteristics of myofibroblasts in liver of sheep naturally infected with the lancet liver fluke (Dicrocoelium dendriticum).

The main objective of our research was to examine the role and immunophenotypic characteristics of myofibroblasts in sheep liver naturally infected by the lancet liver fluke (Dicrocoelium dendriticum). In the reported study we analyzed liver samples from 20 adult sheep, 14 infected animals and 6 controls. The liver samples were fixed in 10% buffered formalin, and routinely processed and stained using hematoxylin eosin, the periodic acid-Schiff and Masson-Goldner trichrome methods. The immunohistochemical examination was carried out by the streptavidin biotin (LSAB2) method, using antibodies for α-smooth muscle actin (α-SMA), desmin and vimentin. The histopathological examination revealed liver fibrosis in 6 out of 14 (42.9%) analyzed samples, while different forms of cholangitis were observed in the remaining 8 out of 14 (57.1%). The expression of α-SMA was proven in perisinusoidal hepatic stellate cells, portal/septal myofibroblasts, and interface myofibroblasts. The degree of α-SMA expression and the number of α-SMA immunopositive cells were the most intensive in the liver with fibrosis. Desmin expression in all liver samples of infected sheep was confirmed in hepatic stellate cells and smooth muscle cells. The hepatic stellate cells, portal/septal myofibroblasts, and interface myofibroblasts reacted as vimentin positive cells. In the liver without fibrotic changes hepatic stellate cells and smooth muscle cells were desmin positive. The obtained results suggest that all populations of myofibroblasts, especially hepatic stellate cells, play an important role in the increased extracellular matrix formation during parasitic liver fibrosis in sheep naturally infected with D. dendriticum.