Evaluation of dietary addition of 2 essential oils from Achillea moschata,or their components (bornyl acetate,camphor, and eucalyptol) on in vitro ruminal fermentation and microbial community composition

This study investigated the effects of 2 Achillea moschata essential oils extracted from plants collected in 2 different valleys of the Italian Alps and 3 pure compounds of oils — bornyl acetate (BOR), camphor (CAM), and eucalyptol (EUCA) — on in vitro ruminal fermentation and microbiota. An in vitro batch fermentation experiment (Exp. 1) tested the addition of all of the substances (2 essential oils and 3 compounds) in fermentation bottles (120 mL) at 48 h of incubation, whereas a subsequent in vitro continuous culture experiment (Exp. 2) evaluated the pure compounds added to the fermenters (2 L) for a longer incubation period (9 d). In both experiments, total mixed rations were incubated with the additives, and samples without additives were included as the control (CTR). Each treatment was tested in duplicate and was repeated in 3 and 2 fermentation runs in Exp. 1 and 2, respectively. Gas production (GP) in Exp. 1 was similar for all of the treatments, and short chain volatile fatty acid (SCFA) production was similar in both experiments except for a decrease of SCFA produced (P = 0.029) due to EUCA addition in Exp. 2. Compared to CTR, BOR and CAM reduced the valerate proportion (P = 0.04) in Exp. 1, and increased (P < 0.01) the acetate proportion in Exp. 2. All treatments increased (P < 0.01) total protozoa counts (+36.7% and +48.4% compared to CTR on average for Exp. 1 and 2, respectively). In Exp. 1, all of the treatments lowered the Bacteroidetes and Firmicutes and increased the Proteobacteria relative abundances (P < 0.05), whereas in Exp. 2, the EUCA addition increased (P = 0.012) the Ruminococcus. In Exp. 1, methane (CH₄) as a proportion of the GP was lowered (P = 0.004) by the addition of CAM and EUCA compared to CTR, whereas in Exp. 2, EUCA reduced the amount of stoichiometrically calculated CH₄ compared to CTR. Overall, essential oils extracted from A. moschata and the pure compounds did not depress in vitro rumen fermentation, except for EUCA in Exp. 2. In both experiments, an increase of the protozoal population occurred for all the additives.     

Midazolam oxidation in cattle liver microsomes: The role of cytochrome P450 3A

In humans, the cytochrome P450 3A (CYP3A) subfamily is involved in midazolam (MDZ) biotransformation into 1′- and 4-hydroxy metabolites, and the former serves as a probe for CYP3A catalytic activity. In veterinary species is still crucial to identify enzyme- and species-specific CYP substrates; thus, the aim of this study was to characterize MDZ oxidation in cattle liver. A HPLC-UV method was used to measure 1′- and 4-hydroxy MDZ (1′- and 4-OHMDZ, respectively) formation in cattle liver microsomes and assess the role of CYP3A by an immunoinhibition study. Moreover, MDZ hydroxylation was evaluated in 300 cattle liver samples and results were correlated with testosterone hydroxylation. Formation of both metabolites conformed to a single-enzyme Michaelis–Menten kinetics. Values of V_max and K_m were 0.67 nmol/min/mg protein and 6.16 μM for 4-OHMDZ, and 0.06 nmol/min/mg protein and 10.08 μM for 1′-OHMDZ. An anti-rat CYP3A1 polyclonal antibody inhibited up to 50% and 94% 1′- and 4-OHMDZ formation, respectively. MDZ oxidation in liver microsomes was poorly correlated with testosterone hydroxylation. In conclusion, cattle metabolized MDZ to 1′-OHMDZ and 4-OHMDZ. The immunoinhibition results indicated a major contribution of CYP3As to 4-OHMDZ formation and the involvement of other CYPs in 1′-OHMDZ production, paving the way for further investigations.     

Correlation between osteoarthritic changes in the stifle joint in dogs and the results of orthopedic,radiographic, ultrasonographic and arthroscopic examinations

Osteoarthritis (OA) is a chronic, degenerative disease affecting the articular cartilage and subchondral bone that causes pain and inhibits movement. The stifle’s joint fibrous capsule contains the synovial membrane, which produces cartilage nutrients. A ruptured cranial cruciate ligament injures the joint and produces OA. Osteoarthritis diagnosis starts with clinical radiographic and ultrasonographic tests, although the latter is not used very much in dog and cat clinics for this purpose. The objective of this study was to establish the correlation among the results of orthopedic, radiographic, ultrasonographic examinations and structural anatomical changes revealed by arthroscopic evaluation to diagnose stifle joint OA and determine risk factors in the dogs affected. Of 44 clinical cases of OA included in the study, 88.64% had ruptured of cranial cruciate ligaments. The correlation between synovial fluid effusion and osteophytosis was of 0.84. It was concluded that there is good diagnostic agreement between synovial fluid effusion and osteophytosis when dealing with stifle joint OA. Risk factors for dogs regarding the development of stifle joint OA included: ruptured cranial cruciate ligaments or patella luxation, female dogs and weight over 10 kg.     

Kinetics of energy source utilization in Boophilus microplus (Canestrini, 1887) (Acari: Ixodidae) embryonic development

The present work evaluates the kinetics of utilization of the main potential energy sources throughout the embryonic developmental stages of Boophilus microplus. The embryonic development of this arthropod is completed in 21 days. Cellularization of the blastoderm occurs on the 6th day and is rapidly followed by germ band extension and segmentation, whose first signs are visible on the 7th day. Cellularization is typically a maternal-driven process, carried out by molecular determinants deposited in the oocyte during oogenesis. On the other hand, segmentation is of zygotic nature, being the consequence of the synthesis of various components by the growing embryo. The enhancement in total B. microplus RNA was observed after cellularization, corroborating the replacement of maternal-driven processes by embryonic zygotic expression. An abrupt increase in oxygen consumption was observed from cellularization until the 8th day of development. The reduction in dry weight at the same period and the susceptibility of oxygen consumption to KCN suggest that the respiration process is activated during early embryonic development. A marked decrease in total lipid content occurred between the 5th and 7th days of development, suggesting this is the main energy source for cellularization. A major reduction in carbohydrate content occurred later, between the 7th and 9th days, and it could be assigned to the morphological segmentation of the embryo. Although the total amount of proteins remains unchanged from oviposition to hatching, a 15% reduction in vitellin (VT) content was observed before cellularization, up to the 4th day after egglaying. This observation was correlated to the synthesis of new proteins needed to support early embryo development. Additional 20% of VT was consumed thereafter, mainly at the end of embryogenesis, and in this case VT is probably used as energy source to the older embryo. Altogether, these data indicate different energy sources for maternal and zygotic driven processes.     

An ovine preimmune foetal model to study the effect of cellular therapies for myocardic diseases

The ovine foetus is an ideal model for preclinical medical studies of cell therapies. It allows us to follow the behaviour of repairing cells inserted into a favourable physiological microenvironment in an animal species more closely related to humans than the rat or rabbit. In addition, the preimmune foetus is able to support cell engraftment by eliminating the problems of tissue rejection. Labelled fibroblasts were transplanted into the myocardium of preimmune foetuses, injecting through a disposable bowed mouth pipette into the left ventricular apex. Two weeks later, foetuses were isolated by laparotomy and each heart was collected and morphologically analyzed. No cases of abortion or postoperative complications in mothers or foetuses occurred. Macroscopically, the isolated hearts did not display any abnormality apart from a small petechia at the injection site. Tissue sections did not show any sign of acute tissue inflammation and viable labelled cells were easily identified among myocardiocytes. This model system guarantees a reduced damage in the engrafted tissue, a high survival and easy retrieval of the injected cells. The direct injection of cells into the preimmune ovine foetus myocardium can be satisfactorily performed to control tissue delivery and to reduce the risk of cell loss and dispersion.     

Clinical and Laboratory Findings in Equine Piroplasmosis

The objective of this study was to evaluate equine piroplasmosis (EP) as a cause of morbidity in horses in Sardinia (Italy), describe the clinical signs and altered hematologic and biochemical parameters, and illustrate response to different treatments. Among 44 horses suspected of tick-borne disease, 38 were polymerase chain reaction (PCR) positive for Theileria equi (n = 27) or Babesia caballi (n = 6), whereas five were positive for both protozoans. Typical clinical features of piroplasmosis were seen in some of the horses, whereas others had nonspecific mild symptoms. Hematologic findings revealed involvement of the three blood cell lineages (anemia, leukopenia or leukocytosis, thrombocytopenia), and biochemical variations were related to increased bilirubin, alteration of serum phosphorus, and hypoalbuminemia. We suggest that the two protozoans are the most important causative agents of equine tick-borne disease in this geographic area, and we observe that different clinical features are associated with the disease; in addition to the typical aspects of piroplasmosis, characterized by fever, pale mucous membranes, and icterus, we can signal other nonspecific mild signs such as weight loss, weight loss associated with an insignificant leukopenia, or weight loss associated with depression, anorexia, and mild hyperbilirubin. The study is intended as a practical contribution for veterinary practitioners because it describes different clinical presentations and laboratory findings of EP, suggests diagnostic and therapeutic approaches to the disease, and shows diffusion of the disease in a Mediterranean region.     

Middle ear cholesteatoma in 11 dogs

Middle ear cholesteatoma is a rare condition in dogs with chronic otitis. Otorrhea, otodinia, and pain on temporomandibular joint palpation are the most common clinical signs. Neurological abnormalities are often detectable. Computed tomography reveals the presence of an expansive and invasive unvascularized lesion involving the tympanic cavity and the bulla, with little or no contrast enhancement after administration of contrast mediu. Video-otoscopy may detect pearly growth or white/yellowish scales in the middle ear cavity. Surgery is the only therapy but is associated with a high risk of recurrence.     

Nuclear remodelling of in vitro grown sheep oocytes

Veterinary Research Communications -