Immunostimulatory and Antibacterial Effects of Cannabis sativa L. Leaves on Broilers.

The aim of this study was to evaluate the effect of dried Cannabis sativa L. leaves as a phytogenic mixture added to broiler feed on CD4+ and CD8+ T lymphocyte subpopulations, Newcastle disease virus (NDV) antibody titres, and the presence of E. coli in faecal samples. The study was conducted on 100 male Ross 308 broilers, divided into four groups of 25 broilers, for a 42-day research period. The groups were housed separately in boxes on a litter of softwood shavings and were fed starter mixture from day 1 to day 21 and finisher mixture from day 22 to day 42. Industrial hemp (C. sativa) was grown in the Crkvina area, Croatia (latitude: 45°18'46.8″ N; longitude: 15°31'30″ E). The hemp leaves were manually separated, sun-dried, and ground to a powder. The mixture offered to the control group did not contain cannabis leaves, whereas the three experimental groups received mixtures containing mixed cannabis leaves in a quantity of 10 g/kg, 20 g/kg, or 30 g/kg (E_10, E_20, and E_30, respectively). The mean NDV antibody level was uniform in all study groups until post-vaccination day 14 and increased comparably with time. The percentage of CD4+ and CD8+ lymphocytes in the peripheral blood subpopulation showed statistically significant differences (p < 0.001) in the E_20 group as compared with the control group and both the E_10 and E_30 groups throughout the study period. As the broiler age increased, the CD4+-to-CD8+ ratios also increased and were statistically significant (p < 0.0001) on day 42 in all experimental groups as compared to the control group. Comparing the control group with the experimental groups indicated that the bacterial count was lower in broiler groups having received feed with the addition of 20 g/kg and 30 g/kg C. sativa leaves. In conclusion, the C. sativa leaves were found to elicit a favourable immunomodulatory effect on cell-mediated and humoral immune responses in broilers via increased CD4+ and CD8+ lymphocyte subpopulations and higher CD4+:CD8+ cell ratios, thus indicating enhanced immune function capacity. In addition, C. sativa leaves may have complementary effects on the broiler post-vaccination immune response, increase broilers' resistance to infectious diseases, reduce the effect of stress associated with vaccination, and improve broiler health and welfare.

Estimation of Population-Specific Genetic Parameters Important for Long-Term Optimum Contribution Selection-Case Study on a Dairy Istrian Sheep Breed.

The Istrian sheep breed has been subjected to selection for dairy traits for more than two decades. However, a detailed study of some important population-specific parameters such as effective population size (Ne) and connectedness between flocks has never been carried out. The aim of the study was to examine the above parameters in dairy Istrian sheep subjected to a national selection program. The Ne was estimated as the mean rate of increase in coancestry, and connectedness was determined using four different statistics. The Ne was estimated at 73 animals with pedigree constraints imposed on 4 equivalent generations and 3 full generations. Analysis of ΔNe ("sliding window approach") revealed a negative ΔNe indicating a progressive loss of genetic variability (ΔNeNEG≥4 = -6.6, p < 0.01; ΔNeNFG≥3 = -4.9, p > 0.05). The overall connectedness (r¯ ~ 0.0001) was below the acceptable level for unbiased ranking of the animals belonging to different flocks (ri,j  = 0.05). OCS appears to be the best option for the long-term survival (self-sufficiency) of the breed, but genetic links between flocks need to be strengthened to allow unbiased ranking of the animals based on the estimated breeding values.

The Impact of Age at First Lambing on Milk Yield and Lactation Length in a Population of Istrian Sheep under Semi-Intensive Management.

This study aimed to examine the impact of ewe's age at first lambing (AFL) on days in milk (DIM), average daily milk yield (DMY), and total milk yield (TMY). Symmetrical bimodal distribution of AFL enabled classification of maidens in those mated in the first (47%) or second year of life (53%). After accounting for all available sources of phenotypic variability with the linear mixed model for repeated records, it was estimated that AFL had a statistically significant effect only on DIM (p < 0.001). The litter size had a significant effect only on TMY (p < 0.001), while the effect of the parity was significant for all the examined traits (p < 0.001). The results of the study suggest that prolongation of age at first mating to the second year of life is not justified in dairy-orientated sheep farms. However, more evidence on this issue is needed for generalization, especially considering some other traits that can impact profitability of dual-purpose sheep farms (reproduction traits, growth rate of lambs, etc.).

Drip loss assessment by EZ and bag methods and their relationship with pH value and color in mutton.

Drip loss, pH value, and color are among the important traits that determine meat quality. Contrary to pH and color, the method associated with drip loss is not yet standardized, and literature data are difficult to compare. Besides, to our knowledge, there is no research comparing drip loss methods and their relation with pH and color in mutton. This study aimed to assess drip loss measurements in mutton taken by different methods (EZ and bag - BM) and their relationship with pH values and color. Mutton samples (Musculus longissimus thoracis et lumborum) originating from 20 ewes of Istrian sheep were used to examine the effect of the method on drip loss after 24 h (EZ 24 vs. BM 24 ) and 48 h (EZ 48 vs. BM 48 ). Furthermore, correlations between drip loss, pH value, and color were analyzed. The statistical analysis was conducted in R programming environment by using different packages. Within the EZ method there was no significant difference ( p > 0.05 ) between ventral and dorsal sample cores used for the assessment of EZ drip loss. Drip loss measured with the same method at two different points of time (24 and 48 h) differed significantly ( p < 0.001 ). There was also a significant difference in drip loss determined by different methods (EZ vs. BM) at the same point of time. There were significant ( p < 0.05 ) correlations between pH 45 min and all color parameters ( L * 4 , a * , b * ). The L * , a * , and b * parameters were highly correlated ( p < 0.001 ). The strongest correlation occurred between a * and b * parameter ( r = 0.93 ). Correlations between drip loss by EZ method and other meat quality attributes were low and not significant. The b * parameter correlated with BM 24 ( r = 0.46 ) and BM 48 ( r = 0.58 ), while a * correlated only with BM 48 ( r = 0.50 ). The correlations between the EZ 24 and BM 24 as well as between the EZ 48 and BM 48 were both non-significant ( p > 0.05 ). Drip loss cannot be predicted with sufficient accuracy by using pH and color. EZ and BM method in mutton do not provide equivalent results for measuring drip loss. Comparisons of the results obtained with different methods should be avoided or at least performed with great precaution.

137Cs in mushrooms from Croatia sampled 15-30 years after Chernobyl.

The aim of this study was to select species with higher potential to accumulate 137Cs among the available mushroom species, by determining the activity concentrations of 137Cs in mushrooms collected along north and north-western part of Croatia. A total of 55 samples of 14 different species were analyzed and the potential of mycorrhizal and saprotrophic species to accumulate 137Cs was compared. A wide range of the dry weight activity concentrations of 137Cs was detected, ranging from 0.95 to 1210 Bq/kg (154 Bq/kg mean value; 52.3 Bq/kg geometric mean) in mycorrhizal and 1.05-36.8 Bq/kg (8.90 Bq/kg mean value; 5.49 Bq/kg geometric mean) in saprotrophic species. Statistical analyses showed that mycorrhizal species accumulate significantly higher concentrations of 137Cs and thus could perform better as long-term bioindicators of environmental pollution by radiocaesium then saprotrophic species. The comparison of Boletus sp. and Hydnum repandum (both mycorrhizal species commonly found in Croatia) showed, in general order of magnitude, higher accumulation in Hydnum repandum. Clearly, mushrooms, especially mycorrhizal species, can be used as significant indicators even decades after the occurrence of any serious 137Cs contamination event. However, as a wide range of values indicates that various parameters may influence the total uptake of the 137Cs into the mushroom fruit bodies, it is necessary to emphasize that 137Cs activity detected in a single mushroom sample is very site-specific.

Lead, cadmium and mercury contents and bioaccumulation potential of wild edible saprophytic and ectomycorrhizal mushrooms, Croatia.

Lead (Pb), cadmium (Cd), and mercury (Hg) contents in ten species of edible mushrooms in Trakoscan, Croatia were determined. In addition, the similarity between the studied species was determined by cluster analysis. The caps and stipes of the fruiting bodies were analysed separately. The analyses were carried out by inductively coupled plasma - optical emission spectrometry (ICP-OES). The greatest mean lead concentrations of 1.91 and 1.60 mg kg -1 were determined in caps and stipes of Macrolepiota procera. The greatest mean concentrations of cadmium (3.23 and 2.24 mg kg-1) were determined in caps and stipes of Agaricus campestris and of mercury (2.56 and 2.35 mg kg-1) in Boletus edulis. In terms of the anatomical parts of the fruiting body (cap-stipe), a considerably greater concentration of the analysed elements was found in the cap for all mushroom species. According to calculated bio-concentration factors, all the examined species were found to be bio-accumulators of Cd and Hg. On the basis of the accumulation of the studied metals, great similarity of mushroom species belonging to the same genus and partial similarity of species of the same ecological affiliation was obtained by cluster analysis.

Heavy metal bioaccumulation by wild edible saprophytic and ectomycorrhizal mushrooms.

Heavy metals cause serious problems in the environment, and they can be accumulated in organisms, especially in the higher fungi. The concentration of Ni, Cr, Pb, Cd, and Hg in 10 species of edible mushrooms in Medvednica Nature Park, Croatia was therefore determined. In addition, the similarity between the studied species was determined by cluster analysis based on concentrations of the aforementioned metals in the fruiting bodies. The contents of nickel, chromium, lead, cadmium, and mercury in the fruiting bodies of mushrooms were obtained by X-ray fluorescence spectrometry. The highest concentrations of Ni (3.62 mg kg(-1)), Cr (3.01 mg kg(-1)), and Cd (2.67 mg kg(-1)) were determined in Agaricus campestris. The highest concentration of Pb (1.67 mg kg(-1)) was determined in Macrolepiota procera, and the highest concentration of Hg (2.39 mg kg(-1)) was determined in Boletus edulis. The concentration of all heavy metals significantly differed (p < 0.001) between examined saprophytic and ectomycorrhizal mushrooms. Considering anatomical part of the fruiting body (cap-stipe), a considerably higher concentration of the analyzed elements was found in the cap for all mushroom species. According to calculated bioconcentration factors, all the examined species were found to be bioexclusors of Ni, Cr, and Pb and bioaccumulators of Cd and Hg. Cluster analysis performed on the basis of the accumulation of the studied metals revealed great phenotypic similarity of mushroom species belonging to the same genus and partial similarity of species of the same ecological affiliation.