Culture of Differentiated Adult Rabbit Auricular Chondrocytes

Chondrocytes dedifferentiate to a fibroblast‐like phenotype on plastic surfaces. Dedifferentiation is reversible if these cells are then cultured embedded in gels as alginate, agarose or collagen. Chondrocytes cultured in suspension on a non‐adherent surface are also known to form aggregates of differentiated cells. The knowledge of chondrocyte behavior in culture is relevant for tissue engineering purposes. In this report we describe a simple method to culture differentiated or redifferentiated rabbit auricular chondrocytes on plastic surfaces with a stable phenotype. When chondrocyte aggregates formed in suspension are next seeded on plastic surfaces, most of them attach to the plastic as round or polygonal cells, and this morphological differentiation, confirmed by the presence of type II collagen, is stable for long culture periods. We also report that the addition of aggregates to monolayer cultures of dedifferentiated chondrocytes results in their redifferentiation, as is shown by their morphological changes and the synthesis of type II collagen. Therefore, this simple method can be useful for the study of chondrocyte behavior on plastic surfaces and for redifferentiating previously proliferated chondrocytes in tissue engineering techniques. Furthermore, these results demonstrate that, in addition to culture conditions such as cell isolation method or cell‐density, chondrocyte behavior on plastic depends on the presence or absence of aggregates resulting from the dissociation process.     

Whole body vibration affects the cross‐sectional area and symmetry of the m. multifidus of the thoracolumbar spine in the horse

Whole body vibration (WBV) has been used as an adjunctive therapy to improve the strength and size of paraspinal muscles as well as postural control in people with lower back pain. It has been proposed that activation of the m. multifidus plays a key role. As the function and anatomy of the m. multifidus in the horse is comparable to that in man, the authors investigated whether WBV might also be a valuable physiotherapeutic modality in horses. The effects of WBV on the cross‐sectional area (CSA) and left to right symmetry of the m. multifidus at various locations of the thoracolumbar spine of the horse was evaluated in a single‐subject quasi‐experimental time‐series design with repeated measure. Nine horses were subject to WBV, 30 min, twice daily, 5 days a week, for 60 days in addition to their regular exercise routine. The CSA of the left and right m. multifidus was measured ultrasonographically at four levels (T15–T16, T16–T17, T18–L1 and L1–L2) along the thoracolumbar spine at Days ?30, 0, 30 and 60 of the study. Changes in the CSA and CSA symmetry (left to right) of the m. multifidus were analysed using nonparametric, repeated measures, comparison of mean ranks with post‐hoc analysis as necessary. A significant increase (P<0.05) in m. multifidus CSA was found at all spinal levels after 30 and 60 days of WBV and a statistically significant improvement in m. multifidus symmetry (becoming more symmetrical) was found after 60 days of WBV, indicating that WBV may be a valuable alternative to dynamic mobilisation exercises when an increase in size and improvement in left to right symmetry of the m. multifidus is sought.     

Influence of different supplements and sugarcane (Saccharum officinarum L.) cultivars on intake, digestible variables and methane production of dairy heifers under tropical conditions

The sulphur hexafluoride (SF(6)) gas tracer method was used to measure methane (CH(4)) production of crossbred (3/4 Holstein x Zebu) dairy heifers fed two types of sugarcane (Saccharum officinarum L.; cultivar IAC-862480 (CC1) or cultivar IAC-873184 (CC2)) and supplemented with urea or concentrate. The study was performed at Embrapa Southeast Cattle, S?o Carlos, SP, Brazil, using a completely randomised design. Differences between treatments were significant for digestibility of dry matter, organic matter and energy. When animals were supplemented with urea differences between sugarcane cultivars did occur for NDF consumption, but not for daily methane production. This suggest that variation in chemical composition of sugarcane did not affect bovine ruminal CH(4) emissions. Concentrate inclusion in animal diet increased digestible organic matter intake, improving the nutrient intake by animals, but did not reduce CH(4) production expressed as a percentage of gross energy intake.     

Seasonal and dietary effects on Vitamin D deficiencies detected in wild boar from mid‐western Spain

Vitamin D (VitD) is involved in important mammalian physiological mechanisms, such as Ca–P metabolism, bone development and immunological response. VitD deficiencies are frequently detected in domestic animals and related to various health problems (e.g., rickets, bone deformation). However, knowledge about the status of VitD in wildlife species, such as the wild boar, is scarce. The aims of this work were to explore VitD status in wild boar populations from mid‐western Spain and to elucidate the influence of daylight exposure and food supplementation in levels of VitD. Serum concentration of VitD (measured as 25‐hydroxivitaminD) was assessed in 276 wild boar from 27 game estates located in mid‐western Spain using a commercial ELISA kit. In 19 out of 27 estates, the staff supplied a specific VitD‐enriched food (2,000 UI/Kg) ad libitum throughout the year, while in the remaining estates (8), no food was supplied. Blood samples were extracted from hunted animals (198) between October and February of hunting seasons 2016/2017 and 2017/2018, and from live wild boar (78) that were captured, sampled and released (March–September of 2017). The percentage of animals with VitD deficiency (<20 ng/ml), VitD insufficiency (20–30 ng/ml) and VitD sufficiency (>30 ng/ml) was estimated, and the relationship of these levels to factors like sex, age and season was assessed using chi‐square tests. Furthermore, associations between daylight exposure and supplemental food with VitD levels were explored using linear models. Of the studied wild boar population, 82.2% showed a VitD deficiency or insufficiency. VitD deficiencies were more frequent in animals sampled in winter and spring. Furthermore, levels of VitD positively correlated with daylight exposure and supplemental food intake. Ad libitum supplementation with VitD‐enriched food was insufficient to prevent VitD deficiencies in wild boar from November to April, probably because food consumption is lower during this period.     

In ovo injection of branched‐chain amino acids: Embryonic development,hatchability and hatching quality of turkey poults

In this study, the influence of a branched‐chain amino acid blend (BCAA composed of 3 l ‐leucine:1 l ‐valine:2 l ‐isoleucine) injected into the amniotic fluid was evaluated for embryonic growth, yolk‐sac (YS) utilization and development of gastrointestinal tract (GIT) and skeletal muscles of turkey embryos from day 24 of incubation (24E) to hatching, together with hatchability, poult quality and liver L* (lightness), a* (redness) and b* (yellowness) values at hatch. At day 22 of incubation, embryonated eggs (n = 240) were assigned to three treatments, that is, eggs were not injected (control, NC) or injected with 1.5 ml sterile solution with 0.9% salt (SA) or 0.2% BCAA blend (BCAAb). These solutions were injected manually into the amniotic fluid of the embryonated eggs. To determine weights and lengths (where appropriate) of the studied organs and tissues, four embryonated eggs and poults per treatment were selected at 24E and at hatch. While the BCAAb decreased the YS and embryo weight, hatchability and the liver L* value, it increased the weight and quality of poults and the weights of breast and thigh muscles at hatch. In conclusion, the in ovo feeding of the BCAA blend negatively affected hatchability but positively affected hatching weight and poult quality by improving development of skeletal muscles and by regulating energy metabolism.     

Seasonality in Saint Croix male lamb reproductive development in northern Mexico

The aim of this study was to determine the influence of the season of birth on the body weight, testicular size, testosterone concentrations, semen output and sexual behaviour in Saint Croix male lambs. Thirty‐three lambs were born in spring (n = 10), summer (Sum; n = 12) or autumn (n = 11). Body weight and the scrotal circumference were weekly recorded; semen was also weekly collected by electroejaculation; and sexual behaviour towards oestrous ewes was weekly evaluated. Data were recorded from 15 to 33 weeks of age. The reproductive development of Saint Croix male lambs differed according to the period in which they were born. More lambs born in spring than in summer and autumn achieved puberty during this study and were also more precocious in semen production and displayed sexual behaviour earlier. Lambs born in summer delayed their semen production; however, season‐related negative effects were less pronounced at the end of the study than in autumn born lambs born. In conclusion, despite the apparently low seasonality of the breed, Saint Croix male lambs showed in this study a seasonal pattern in their reproductive development at least until they achieve the puberty.     

Follicular dynamics and changes in oestradiol‐17β, progesterone and LH profiles following PGF2α induced oestrus in early and late ovulating Beetal goats

This study compares the factors associated with variable interval to oestrus and ovulation between early versus late ovulating goats following PGF_2α administration. The time of ovulation in Beetal goats (n = 38) was monitored through transrectal ultrasound at every 6 hr following a single dose of PGF_2α (experiment 1). Variations in oestrus and ovulation times were further explored through the changes in follicular dynamics, endocrine profiles and behaviour in another set of goats (n = 13) following single PGF_2α given randomly during the luteal phase (experiment 2). The ovulation time varied between 60 and 96 hr, and 57% of ovulations occurred by 72 hr following PGF_2α (experiment 1). Accordingly, the goats (n = 13) in the second experiment were retrospectively divided either into early and/or late ovulating, that is, ≤72 and/or ≥84 hr following PGF_2α. The onset of oestrus, peak estradiol‐17β concentration and LH surge after PGF_2α was first observed in early than late ovulating goats (p < 0.05). The goats ovulating early had larger follicle and smaller CL in diameter at the time of PGF_2α administration than those ovulating late (5.4 ± 0.2 vs. 4.3 ± 0.2 mm and 10 ± 0.6 vs. 11.8 ± 0.3 mm, respectively; p < 0.05). Likewise, plasma progesterone concentration tended to be lower (p = 0.087) in early than late ovulating goats. In conclusion, the size of dominant follicle and CL at the time of PGF_2a determines the interval to ovulation following a single dose of PGF_2a during the luteal phase.