Učinak eksrakta kore brucijskog bora (Pinus brutia) na čiste i mješovite kulture buražnih bakterija i arheja te na fermentacijske značajke buraga in vitro

The aim of the study was to investigate the effects of Pinus brutia bark extract, which is rich in polyphenolic compounds of tannins, on both pure and mixed continuous cultures of rumen bacteria and archaea, as well as on rumen fermentation characteristics in vitro. Antimicrobial susceptibility assay with pure cultures was carried out in an anaerobic chamber. Pinus brutia bark extract exhibited a potential inhibitor activity (P<0.05) against pure cultures of Ruminococcus flavefaciens, Eubacterium ruminantium, and Methanobacterium formicicum while a growth stimulatory effect (P<0.05) was observed for Ruminoccocus albus, Butyrivibrio fibrisolvens, and Streptococcus bovis. Pinus brutia bark extract only had a potential inhibitor effect (P<0.05) on R. albus at the highest dose (1200 μg/mL). Pinus brutia bark extract also stimulated (P<0.05) the growth of pure cultures of Fibrobacter succinogenes, while it did not affect Megasphaera elsdenii, except at the highest dose. The effects of two doses (75 and 375 mg/L) of P. brutia bark extract on in vitro mixed cultures and rumen fermentation parameters were determined by the rumen simulation technique (Rusitec). Supplementation with P. brutia bark extract led to a quadratic decrease (P<0.05) in the cell numbers of R. flavefaciens. Production of total and individual short chain fatty acids (SCFA), acetate to propionate ratio (C2/C3), total protozoa, ruminal pH, and dry matter digestibility (DMD) did not change in the presence of P. brutia bark extract. Supplementation with both doses of P. brutia bark extract decreased (P<0.05) the ammonia-N concentrations. Ammonia-N concentration was lowest in the high-supplemented group (P<0.05). As a conclusion, inhibitory effects of P. brutia bark extract on some species in the pure cultures were in the same direction as with mixed ruminal cultures, while stimulatory effects disappeared. The lack of inhibitory effects on protozoa and on a large number of Gram-positive rumen bacteria in the mixed cultures suggests that its mechanism of action is not exactly similar to antibiotics. Although P. brutia bark extract did not alter ruminal SCFA, it could have potential to improve ruminal protein utilization without depressing rumen microbial fermentation.Cilj ovog rada bio je istražiti učinak ekstrakta kore brucijskog bora (Pinus brutia), koji je bogat polifenolnim sastojcima tanina, na čiste i mješovite kulture buražnih bakterija i arheja kao i na in vitro fermentacijske značajke buraga. Proveden je test antimikrobne osjetljivosti s čistim kulturama u anaerobnim uvjetima. Ekstrakt kore brucijskog bora pokazao je potencijalnu inhibitornu aktivnost (P < 0,05) protiv čistih kultura bakterija Ruminococcus flavefaciens, Eubacterium ruminantium i Methanobacterium formicicum, a stimulacijski učinak na rast (P < 0,05) opažen je za bakterije Ruminoccocus albus, Butyrivibrio fibrisolvens i Streptococcus bovis. Ekstrakt kore brucijskog bora imao je potencijalan inhibitorni učinak (P < 0,05) na R. albus samo u najvećoj dozi (1200 μg/mL). Također je imao stimulacijski učinak (P < 0,05) na čiste kulture Fibrobacter succinogenes, a nije utjecao na Megasphaera elsdenii osim u najvećoj dozi. Učinak dviju doza (75 i 375 mg/L) ekstrakta kore brucijskog bora na in vitro mješovite culture i pokazatelje fermentacije u buragu određen je simulacijskom tehnikom (Rusitec). Dodatak ekstrakta kore brucijskog bora doveo je do kvadratnog smanjenja (P < 0,05) broja stanica R. flavefaciens. Nije bilo promjena u proizvodnji ukupnih i pojedinačnih kratkolančanih masnih kiselina (SCFA), omjeru acetata i propionata (C2/C3), ukupnom broju protozoa, buražnom pH i probavljivosti suhe tvari (DMD). Suplementacija objema dozama ekstrakta kore brucijskog bora smanjila je (P < 0,05) koncentracije amonijaka-N. Koncentracija amonijaka-N bila je najniža u skupini s najvećom dozom suplementa (P < 0,05). Zaključujemo da je inhibitorni učinak ekstrakta kore brucijskog bora na neke vrste u čistim kulturama bio jednak onomu u mješovitim buražnim kulturama, a nije bilo stimulacijskog efekta. Manjak inhibitornih učinaka na protozoe i na mnoge Gram-pozitivne buražne bakterije u mješovitim kulturama upućuje na to da njihov mehanizam djelovanja nije jednak onomu kod antibiotika. Premda ekstrakt kore brucijskog bora nije promijenio buražni SCFA, mogao bi poboljšati iskorištavanje proteina u buragu a da pritom ne suprimira mikrobnu fermentaciju

Acute and chronic stress alter behavioral laterality in dogs

Dogs are one of the key animal species in investigating the biological mechanisms of behavioral laterality. Cerebral asymmetries are assumed to be influenced by stress, but this subject has not yet been studied in dogs. This study aims to investigate the effect of stress on laterality in dogs by using two different motor laterality tests: the $Kong^{TM}$ Test and a Food-Reaching Test (FRT). Motor laterality of chronically stressed (n = 28) and emotionally/physically healthy dogs (n = 32) were determined in two different environments, i.e., a home environment and a stressful open field test (OFT) environment. Physiological parameters including salivary cortisol, respiratory rate, and heart rate were measured for each dog, under both conditions. Cortisol results showed that acute stress induction by OFT was successful. A shift towards ambilaterality was detected in dogs after acute stress. Results also showed a significantly lower absolute laterality index in the chronically stressed dogs. Moreover, the direction of the first paw used in FRT was a good predictor of the general paw preference of an animal. Overall, these results provide evidence that both acute and chronic stress exposure can change behavioral asymmetries in dogs