Postpartum oestrous cycling resumption of yak cows following different calf weaning strategies under range conditions

Early weaning can improve body condition and reproductive performance of cows. The objectives of this study were to examine and compare oestrous cycling resumption, behaviour and blood parameters of yak cows following four different strategies of calf weaning. Twenty‐six yak cows (4–8 years) and their calves (94.3 ± 2.4 days) were studied in which calves were: weaned naturally with free access to their mothers (NW; n = 13); weaned abruptly and separated permanently from their mothers (PW; n = 5); separated temporarily from their mothers for 15 days (TW; n = 5); and fitted with nose plates for 15 days, but allowed free access to their mothers (NP; n = 3). Yak cows with sucking calves (NW) spent more time grazing in the cold season (from d 39 to 84). Based on serum progesterone concentrations, none of nine NW yak cows resumed oestrous cycling during the study, while seven of nine yak cows with weaned calves resumed cycling (3/3 for PW; 2/3 for TW; and 2/3 for NP yaks). We concluded that early weaning improved postpartum reproductive performance of yak cows on the Qinghai‐Tibetan plateau and that abrupt and permanent weaning was an appropriate strategy.     

Proteomic identification of ruminal epithelial protein expression profiles in response to starter feed supplementation in pre-weaned lambs

The present study aimed to comparatively characterize the ruminal epithelial protein expression profiles in lambs fed ewe milk or milk plus starter diet using proteome analysis. Twenty new-born lambs were randomly divided into a group receiving ewe milk (M, n = 10) and a group receiving milk plus starter diet (M + S, n = 10). From 10 d old, M group lambs remained with the ewe and suckled ewe milk without receiving the starter diet. The lambs in the M + S group were separated from the ewe and received starter feed. All lambs were slaughtered at 56 d old. Eight rumen epithelia samples (4 per group) were collected to characterize their protein expression profiles using proteomic technology. Proteome analysis showed that 31 upregulated proteins and 40 downregulated proteins were identified in the rumen epithelium of lambs in response to starter diet supplementation. The results showed that starter feeding regulates a variety of biological processes in the epithelium, especially blood vessel development and extracellular matrix protein expression. Meanwhile, the expression of proteins associated with synthesis and degradation of ketone bodies, butanoate metabolism, and citrate cycle signaling transduction pathway were upregulated in the group with starter diet supplementation, including 3-hydroxy-3-methylglutaryl coenzyme A synthase (HMGCS2, fold change [FC] = 1.93), 3-hydroxybutyrate dehydrogenase 1 (BDH1, FC = 1.91), and isocitrate dehydrogenase 1 (IDH1, FC = 8.12). The metabolic processes associated with ammonia detoxification and antioxidant stress were also affected by starter diet supplementation, with proteins, microsomal glutathione S-transferase 3 (MGST3, FC = 2.37) and IDH1, linked to the biosynthesis of glutamate and glutathione metabolism pathway being upregulated in the group with starter diet supplementation. In addition, starter feeding decreased the expression of Ras-related protein rap-1A (RAP1A, FC = 0.48) enriched in Rap1 signaling pathway, Ras signaling pathway, cyclic adenosine monophosphate (cAMP) signaling pathway, and mitogen-activated protein kinase (MAPK) signaling pathway. In summary, starter feed supplementation changed the expression of proteins related to energy production, ammonia detoxification, antioxidant stress, and signaling pathways related to proliferation and apoptosis, which facilitates the rumen epithelia development in lambs. The results provide new insights into the molecular adaptation of rumen epithelia in response to starter diet supplementation at the protein level in lambs.     

Luteal vascularity and embryo dynamics in mares during early gestation: Effect of age and endometrial degeneration

The present study characterized the luteal status and the dynamic of the conceptus during the first 20 days of gestation in mares with different ages and degrees of endometrial degeneration. Total area of the corpus luteum (CL), luteal vascularity, CL area with blood signals, progesterone concentrations (P4), embryonic vesicle diameter, number of embryonic location changes, embryonic fixation position and uterine contractility were evaluated. In Experiment 1, mares ≤6 years of age (Young group, 5.6 ± 0.2 years, n = 7 mares) and mares ≥15 years of age (Old group, 17.2 ± 0.9 years, n = 6 mares) were used to investigate the effect of age. In Experiment 2, the luteal and embryonic parameters were compared between mares with minimal (Mild group, endometrial category I, n = 9 mares) and severe (Severe group, endometrial category III, n = 7 mares) endometrial degeneration. The Old and Severe groups had greater (p ≤ 0.04) total CL area and reduced luteal vascularity (p ≤ 0.04) than the Young and Mild groups, respectively. However, P4 levels and CL area with blood signals were similar (p ≥ 0.8) between the groups. A negative effect of age (p < 0.01), but not of endometrial degeneration (p = 0.6), was found for the embryonic vesicle diameter. The conceptus mobility was high (p > 0.1) until day 14 of gestation in the Severe group, while a reduced number of changes of the embryo location was detected earlier (p < 0.05) in the Old group. In conclusion, the newly formed CL of aged mares and mares with severe endometrial degeneration suffered a structural remodelling to safeguard the local blood supply and the continuous P4 output during early gestation. Moreover, an earlier reduction of the embryonic mobility and a delayed development of the conceptus were associated with advanced age, regardless of the degree of endometrial degeneration.     

The effect of maternal obesity on fatty acid transporter expression and lipid metabolism in the full‐term placenta of lean breed swine

This study was conducted to evaluate the influence of back‐fat thickness (BF), at mating of sows, on the maternal and newborn circulating lipids, expression of placental fatty acids (FA) transporters and lipid accumulation in placenta. Full‐term placentas were obtained by vaginal delivery from BFI (9–14 mm; n = 37), BFII (15–19 mm; n = 43) and BFIII (20–27 mm; n = 38) sows according to BF at mating, and frozen placental sections were analysed for fat accumulation. Blood samples were collected from the sows of day 105 pregnancy and from cord blood at delivery. mRNA and protein expression levels were evaluated with real‐time RT‐PCR and Western blotting. Our results demonstrated that BFII females had significantly increased litter weight and placental efficiency, decreased maternal triglyceride (TG) and non‐esterified fatty acids (NEFA) levels, decreased maternal IL‐6, TNFα and leptin levels compared to BFIII females (p < .05). BFIII sows were associated with significantly decreased newborn TG levels, increased newborn glucose, IL‐6 and TNFα levels compared to BFI or BFII sows (p < .05). BFI and BFII females had significantly decreased placental TG, NEFA and cholesterol (CHOL) contents compared to BFIII females (p < .05). Moreover, decreased CD36, FATP1, FABP4, and FABP1 mRNA and protein and FATP4 protein expression, and increased LPL activity were also observed in BFIII group compared with BFII group (p < .05). PPARγ mRNA and protein and lipogenic genes such as SREBP‐1c, ACSL1, ACCα, FAS and SCD mRNA expression were downregulated or upregulated, respectively, in the placentas of BFIII sows compared to BFI or BFII sows (p < .05). Overall, this study demonstrated that there is no advantage, in terms of litter live size, litter weight and placental FA transport and metabolism, in performing the mating of sows with BF>19 mm.     

Reproductive Development of Santa Inês Rams During the First Year of Life: Body and Testis Growth,Testosterone Concentrations,Sperm Parameters,Age at Puberty and Seminal Plasma Proteins

We have investigated the reproductive development of the tropically adapted Santa Inês ram, the most common hair sheep in Brazil. From 8 to 48 weeks of age, 16 animals were evaluated for body and testis growth, semen parameters, testosterone concentrations and seminal plasma proteins, using two‐dimensional SDS‐PAGE. Animals were weaned at 30 days and kept in feedlots thereafter, receiving hay, concentrate (18% of crude protein) and mineral supplement. Body weight increased from 12.3 ± 0.7 to 54.3 ± 1.6 kg between 8 and 48 weeks (p < 0.05), but changes in thoracic perimeter and scrotal circumference were non‐significant after 36 weeks (p > 0.05). The percentage of motile sperm increased slowly until 23 weeks and more rapidly after that age, but significant changes in progressive motility occurred after 25 weeks. Presence of abnormal sperm related inversely to age. Most significant changes in sperm concentration occurred between 38 and 44 weeks (0.38 ± 0.05 to 1.14 ± 0.24 × 10⁹ cells/ml, p < 0.05) and testosterone reached its highest concentrations at 42 weeks, decreasing afterwards. Rams reached puberty at 28.2 ± 0.8 weeks. The number of protein spots on seminal plasma gels was similar from 15 to 18 weeks (45 and 47 spots; p > 0.05), increased until 24 weeks (141 spots) and 28 weeks (170 spots; p < 0.05) and remained without significant (p > 0.05) changes from 28 to 48 weeks (186 ± 10 spots). Furthermore, the intensity of selected spots on 2D maps increased (p < 0.05) between 15 and 28 weeks, which preceded or coincided with the main developmental changes in sperm motility and percentage of defective sperm in the ejaculates. These results will support future studies designed to characterize specific seminal plasma proteins whose expression relate to the development of testis, epididymis and accessory sex glands.     

Endometrial transcript profile of progesterone‐regulated genes during early pregnancy of Water Buffalo (Bubalus bubalis)

Progesterone (P₄) plays a key role in the establishment and maintenance of pregnancy in most mammals. Unravelling the expression of progesterone‐regulated genes can expand the understanding of the embryonic mortality. Accordingly, we studied the relative mRNA expression of the P₄‐regulated genes in the buffalo. Uteri were collected from the abattoir and categorized into nonpregnant late luteal phase, stage I (28–38th days of gestation) and stage II (48–56th days of gestation) of pregnancy (n = 6/group). After extraction of total RNA from the endometrial tissues, we carried out qRT‐PCR for determining the relative mRNA expression of the P₄‐regulated genes using nonpregnant late luteal phase as calibrator group. The expression of LGALS3BP (essential for maternal recognition of pregnancy) gene was found to be significantly upregulated (p < 0.05), while MUC1 (important for embryo attachment) gene was downregulated in stage I and II of pregnancy. We observed no significant change in the expression of LGALS1, LGALS9 and CTSL genes. The SLC5A11 and SLC2A1 genes (involved in the transport of glucose to endometrium) in early pregnancy were upregulated in the pregnancy stage I (p < 0.05) relative to nonpregnant late luteal phase. The CST3 gene was significantly upregulated in pregnancy stage II (p < 0.01). These results provide molecular insights into the specific pathways involved in foeto‐maternal communication during early pregnancy in buffaloes.     

Molecular signature and colony morphology affect in vitro pluripotency of porcine induced pluripotent stem cells

Overall efficiency of cell reprogramming for porcine fibroblasts into induced pluripotent stem cells (iPSCs) is currently poor, and few cell lines have been established. This study examined gene expression during early phase of cellular reprogramming in the relationship to the iPSC colony morphology and in vitro pluripotent characteristics. Fibroblasts were reprogrammed with OCT4, SOX2, KLF4 and c-MYC. Two different colony morphologies referred to either compact (n = 10) or loose (n = 10) colonies were further examined for proliferative activity, gene expression and in vitro pluripotency. A total of 1,697 iPSC-like colonies (2.34%) were observed after gene transduction. The compact colonies contained with tightly packed cells with a distinct-clear border between the colony and feeder cells, while loose colonies demonstrated irregular colony boundary. For quantitative expression of genes responsible for early phase cell reprogramming, the Dppa2 and EpCAM were significantly upregulated while NR0B1 was downregulated in compact colonies compared with loose phenotype (p < .05). Higher proportion of compact iPSC phenotype (5 of 10, 50%) could be maintained in undifferentiated state for more than 50 passages compared unfavourably with loose morphology (3 of 10, 30%). All iPS cell lines obtained from these two types of colony morphologies expressed pluripotent genes and proteins (OCT4, NANOG and E-cadherin). In addition, they could aggregate and form three-dimensional structure of embryoid bodies. However, only compact iPSC colonies differentiated into three germ layers. Molecular signature of early phase of cell reprogramming coupled with primary colony morphology reflected the in vitro pluripotency of porcine iPSCs. These findings can be simply applied for pre-screening selection of the porcine iPSC cell line.     

Factors associated with ewe and lamb survival,and subsequent reproductive performance of sheep undergoing emergency caesarean section

Clinical records of all 212 ewes undergoing emergency caesarean surgery at a veterinary teaching hospital between January 2008 and December 2019 were evaluated retrospectively. Their age ranged from 1 to 10 years (median = 4 years), with German merino the predominant breed (48.1% of cases). The most frequently diagnosed indications were insufficient cervical dilatation (n = 94, 44.3%), uterine torsion (n = 50, 23.6%), foetopelvic disproportion (n = 31, 14.6%) and vaginal prolapse intra partum (n = 11, 5.2%). Fifty-four (25.5%) of the 212 ewes additionally suffered from one or more concurrent, pre-existing conditions. Overall ewe mortality until hospital discharge was 10.8% (23/212), and 3.8% (n = 6) for the 158 ewes without a history of concurrent disorders. Mortality during hospitalization increased to 31.5% (17/54) for those with pre-existing conditions. Total lamb mortality was 49.1% (173/352) until hospital discharge. Pre-existing conditions (p = .001) and the presence of post-surgical complications (p = .025) were identified as significant factors influencing dam mortality, while delayed presentation for veterinary attention with an observed duration of labour of >12 hr was identified as the most influential factor on total lamb mortality (p = .010). The presence of dead or emphysematous foetuses was not significant for ewe mortality. Follow-up information on further outcomes was available for 156 (82.5%) of the 189 discharged ewes. Eighty-nine animals (57.1%) were re-bred in the following season and achieved a 93.3% (83/89) pregnancy rate, while the remainder had either been slaughtered (n = 56, 35.9%), sold (n = 5, 3.2%) or had died of unknown causes (n = 3, 1.9%). The subsequent incidence of dystocia was 15.6% (n = 12) in the 77 ewes with available information on lambing ease. Adequate management of underlying conditions and timely intervention are important factors for best possible short-term outcomes. In the long term, the subsequent pregnancy rate was good and the incidence of subsequent dystocia was within the normal range.     

Proteome of the periovulatory oviduct and uterus of goats as related to nutritional balance

This study was conducted to evaluate the effects of different feeding levels on the proteome of oviduct and uterus tissues of hormonally stimulated goats during the periovulatory period. Forty goats were separated into four different diet groups: Diet 1.0 M (n = 11), Diet 1.3 M (n = 10), Diet 1.6 M (n = 9), Diet 1.9 M (n = 10), fed with 1.0, 1.3, 1.6 and 1.9 times live weight maintenance, respectively. After four weeks of treatment, six hormonally stimulated females per treatment group were randomly selected for collection of uterine and the oviduct tissue samples. Samples were collected after animals were slaughtered in a commercial unit. Feeding goats with 1.3 to 1.9 times more nutrients than a control group directly influenced the proteome of the oviduct and uterus, altering the expression of proteins that participate in biological processes such as apoptosis, antioxidant, and immunological activities. These events are crucial for fertilization and early embryonic survival. Expression of oviduct proteins such as Tubulin Beta 2B, Transferrin and Disulphide‐isomerase A3 increased in the 1.9 M group in relation to the other feeding levels. Disulphide‐isomerase A4 showed higher expression in the 1.0 M group compared to diets with higher energetic levels. As energy intake increased in the diets, there was higher expression of Alpha‐1‐antitrypsin and downregulation of Profilin‐1 in the uterus of the goats. In conclusion, this study showed that specific proteins of the goat oviduct and uterus expressed during the periovulatory period are modified as the result of nutritional balance.     

Molecular cloning and expression of FGF2 gene in pre‐implantation developmental stages of in vitro‐produced sheep embryos

Early embryonic mortality is one of the main sources of reproductive loss in domestic ruminants including sheep. Fibroblast growth factor‐2 (FGF‐2) is a member of FGFs family that mediates trophoblast activities and regulates embryonic development in various species. In this study, we have cloned, characterized sheep FGF2 cDNA (KU316368) and studied the expression in sheep embryos. Ovaries of non‐pregnant sheep were collected from local abattoir and matured in culture medium at 38.5ºC, 5% CO₂, 95% humidity for 22–24 hr. The matured oocytes were inseminated with capacitated spermatozoa in Brackett and Oliphant medium and resulted embryos were cultured in CO₂ incubator for 6–7 days to complete the developmental stages from two cells to blastocyst stage. Total RNA was extracted from immature oocytes (n = 100), mature oocytes (n = 100) and different stages of embryos such as 2 cell (n = 50), 4 cell (n = 25), 8 cell (n = 12), 16 cell (n = 6), morula (n = 5) and blastocyst (n = 3). The total RNA isolated from the oocytes and embryos was reverse transcribed and subjected to real‐time polymerase chain reaction using sequence‐specific primers and SYBR green as the DNA dye. On sequence analysis, the nucleotide sequence of sheep FGF2 exhibited highest sequence similarity with cattle (100%) and least with rat and mouse (69.2%). At the deduced amino acid level, a highest degree of similarity was noticed with cattle, buffalo, goat, pig, camel and horse (100%) and lowest degree of identity with rat, human and mouse (98.2%). The FGF2 mRNA expression was higher in immature and mature oocytes and gradually decreases from 2‐cell stage of embryo to the blastocyst stage. More over a significant differences in FGF2 mRNA expression (p < .05) were observed between immature oocytes and all pre‐implantation stages of embryo. It can be concluded that FGF‐2 plays a significant role in pre‐implantation and early development of embryos in sheep.     

Differential expression of Wilms’ tumour 1 gene in porcine urogenital organs during development

Wilms’ tumour 1 gene (WT1) is essential for the development of mammalian urogenital system. However, the expression pattern of WT1 in the development of porcine urogenital organs is still unclear. Here, we examined the expression of WT1 mRNA and protein in porcine kidneys, ovaries and testes from embryonic days 35 and 60 (E35d, E60d, n = 3) to the newborn (0d, n = 4) and adult (210d, n = 3) stages, using real‐time PCR and immunofluorescent staining. Real‐time PCR analysis showed that porcine kidneys, ovaries and testes all expressed high level of WT1 mRNAs, especially in adult testes (p < 0.05 or 0.01 vs. kidney and ovary, respectively). Morphologically, characteristic microstructures of the kidneys, ovaries and testes were observed and discerned at all four stages. Immunofluorescently, WT1 expression was detected in a dynamic and context‐specific pattern during the development of these organs. Taken together, porcine urogenital organs express relatively high levels of WT1 mRNA. Dynamical and context‐specific expression profile of WT1 in these organs occurs during their development, implying its close association with the development and function of porcine kidney, ovary and testis.     

Absence of Sirtuin 1 impairs the testicular development in cattleyak by inactivating SF-1

Cattleyak, which are interspecific hybrids between cattle and yak, display much higher growth performances than yak. However, F1 male cattleyak are infertile due to defective testicular development. Sirtuin 1 (SIRT1) is a histone deacetylase that is essential for various biological processes, while the roles of testicular SIRT1 in yak and cattleyak are still poorly understood. Here, we found that SIRT1 was localized in various kinds of yak testicular cells except elongated spermatids while it was deficient in cattleyak testis. Further studies indicated that cattleyak testis exhibited decreased histone acetylation levels on H3 and H4. One of SIRT1 co-factors, steroidogenic factor-1 (SF-1), was lost in cattleyak testis at protein level. Expressions of several SF-1 target genes responsible for Sertoli cell development and steroidogenesis, including STAR, CYP11A1, CYP26B1, FDX1 and HSD3B, decreased significantly in cattleyak testis. In addition, SIRT1-mediated P53 acetylation was not responsible for the cell apoptosis in cattleyak testis. Taken together, our results suggested the deficiency of SIRT1 in yak testis caused inactivation of SF-1 and the impairment of testicular development. This research provides theoretical bases for understanding the mechanism of cattleyak sterility and gives new insights in revealing the roles of SIRT1 in regulating yak testicular development.     

Effect of feeding rice whole crop silage on growth rate,levels of vitamin A, β‐carotene,vitamin E and IGF‐1 in plasma and skeletal muscle protein degradation in Japanese black calves

This study evaluated the effects of rice whole crop silage (RWCS) on growth, plasma levels of vitamin A, β‐carotene, vitamin E and IGF‐1, and expression of genes involved in muscle protein degradation and synthesis in Japanese Black calves. Eleven calves were divided into RWCS (fed RWCS ad libitum and concentrate, n = 5) and control groups (fed hay ad libitum and concentrate, n = 6). Final body weight and dairy gain were significantly larger in the RWCS group compared with the control group. Plasma β‐carotene and vitamin E concentrations were significantly higher in the RWCS group compared with control group. Although plasma vitamin E concentration in the RWCS group significantly increased from 4 to 9 months of age, it did not increase in the control group. At 6 months of age in the RWCS group, ubiquitin B (p < 0.05) and calpain 1 (p = 0.097) mRNA expression were lower than control group, but they were not different between groups at 9 months of age. These results indicate that RWCS increases plasma β‐carotene level and promotes muscle growth because of a decrease in the rate of protein degradation, but the effect is lost with the increase in plasma vitamin E level.     

Reduced lung lesions in pigs challenged 25 weeks after the administration of a single dose of Mycoplasma hyopneumoniae vaccine at approximately 1 week of age

Two independent studies assessed the duration of immunity of an inactivated adjuvanted Mycoplasma hyopneumoniae vaccine against mycoplasmal pneumonia in seronegative (study A, n = 52) and seropositive (study B, n = 52) pigs. The pigs were allocated randomly to treatment and were then injected with a single dose of either the vaccine or a placebo at approximately 1 week of age. Twenty-five weeks after treatment administration, the pigs were challenged with a virulent strain (LI 36, Strain 232) of M. hyopneumoniae and the extent of lung lesions consistent with mycoplasmal pneumonia was assessed 4 weeks later.In study A, the geometric mean lung lesion score (expressed as least squares mean percentages of lung lesions) was significantly (P = 0.0001) lower in vaccinated (0.3%, n = 20) than in control pigs (5.9%, n = 24) seronegative to M. hyopneumoniae at enrolment; similarly, in study B, the extent of lung lesions was significantly reduced (P = 0.0385) in seropositive vaccinated pigs (2.0%, n = 22) compared to controls (4.5%, n = 26). At the end of the investigation period, 4 weeks after challenge, mean antibody sample-to-positive (S/P) ratios were significantly higher both in seronegative (P = 0.0012) and seropositive (P = 0.0001) vaccinated pigs (mean values = 0.77 and 0.81, respectively) than in controls (mean values = 0.51 and 0.38, respectively).     

Retrospective Study of Cryptosporidiosis Among Diarrhoeic Children in the Arid Region of North‐Eastern Nigeria

A retrospective study was carried out to investigate the prevalence of Cryptosporidium oocysts among diarrhoeic children (n = 650), aged between 0 and 15 years, living in Maiduguri metropolis (n = 220), Bama local government area (n = 278) and Gwoza local government area (n = 152). Stool samples were concentrated using the ethyl acetate sedimentation method. Data of stool samples with Cryptosporidium oocysts from patients within the specified age groups were collected and analysed. The overall prevalence was 42.9%. The prevalence was higher in Maiduguri metropolis 45.0%, which is an urban area as compared to Gwoza and Bama combined together 41.8% which are rural areas but not statistically significant at 95% confidence level (P > 0.05; OR = 1.14; CI = 0.82, 1.58). According to age, the prevalence in age group A (0–10 years) was higher (46.8%) as compared to age group B (11 < 15 years), which was 20.8%, and this was statistically significant at 95% confidence level (P < 0.05; OR = 3.34; CI = 1.98, 5.61). According to gender, males showed a higher prevalence (52.5%) compared with females (47.5%), but this was not statistically significant at 95% confidence level (P > 0.05; OR = 1.13; CI = 0.82, 1.53). Seasonal prevalence showed that hot dry months of March and April were higher compared with other months. Our findings indicate the presence of the pathogen in children in Borno State, Nigeria, with higher odds of the infection in younger children, and dry months may be more associated with the infection. Control and preventive measures should be taken to protect younger children from the infection.     

Age- and Sex-Related Modifications of Hematology in Spanish Purebred Horse

The purpose of this study was to test the hypothesis that age and sex significantly modulate the changes in hematological parameters of physiologically healthy Spanish Purebred horse, establishing the reference values. Blood samples were obtained on a total of 159 horses (n = 77 males and n = 82 females), classified in four age groups: group A (1–2 years; n = 39), group B (>2–3 years; n = 38), group C (>3–4 years; n = 41), and group D (>4–7 years; n = 41). By means ADVIA 2020i, red blood cell (RBC), hemoglobin concentration (HB), packed cell volume, mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration, platelets (PLTs), as well as total and differential white blood cells (WBCs) were determined. Total plasma protein was determined by spectrophotometry. Results obtained showed that sex had a limited influence on these parameters, being the HB higher in males and WBC higher in females (P = .047). Age caused a reduction in RBC, with a compensatory increase of MCV and MCH, decrease in WBC, PLT, lymphocytes (LYMPs), as well as an increase of N/L ratio (P = .047). Interactions between age and sex showed higher RBC in males than in females of the group B, and higher HB and WBC in males than in females of the group D. According to hematological data, Spanish Purebred horse was classified as a warm- or cold-blooded horse. Decline of RBC, WBC, and PLT parameters is a natural condition of horse with the advance of age, indicating a decrease of the narrow bone response.     

Influence of dietary lysine level on whole-body protein turnover, plasma IGF-I, GH and insulin concentration in growing pigs

Four growing pigs (initial liveweight 25.9 ± 0.54 kg, final liveweight 43.0 ± 1.06 kg) were used to study the effect of dietary lysine level on nutrient digestibility, whole-body protein turnover, plasma insulin-like growth factor-I (IGF-I), growth hormone (GH), insulin, glucose, and urea nitrogen (PUN). Four diets, containing 7.0 g (L1), 9.5 g (L2), 12.0 g (L3) and 14.5 g (L4) lysine per kg diet respectively, were formulated as experimental treatments. The animals and diets were allocated in a 4 × 4 Latin square design. Nitrogen (N) metabolism and whole-body protein turnover were measured by classical method and single-dose ¹⁵N end-product method, respectively. The blood samples were taken at the end of each experimental period. Results showed that N retention (NR) and N biological value (NBV) were significantly increased from L1 to L4 (P < 0.05). However, differences in NR and NBV between L2, L3 and L4 were not significant (P > 0.05). There was no significant difference on dry matter (DM) digestibility, organic matter (OM) digestibility and N digestibility between different treatments (P > 0.05). Whole-body protein synthesis, protein degradation and protein accretion increased markedly from L1 to L2 (P < 0.05), but did not increase further from L2 to L4. Whole-body protein accretion (y, g/kg W^0.75/d) increased with dietary lysine (x, g/kg) in a quadratic manner: y = − 0.09x² + 2.12x − 5.14 (r² = 0.96, n = 4, P < 0.05).The results also showed that differences in plasma IGF-I, GH, glucose and PUN concentration between different treatments were not significant (P > 0.05). Plasma insulin concentration (y, μIU/ml) was increased with dietary lysine (x, g/kg) in a quadratic manner: y = 0.23x² − 4.10x + 32.25 (r² = 0.99, n = 4, P < 0.05), but it was not found that plasma insulin concentration was related to NR. A significant correlation was found between NR (y, g/d) and plasma IGF-I (x, ng/ml): y = − 3.1 × 10^− 3x² + 1.31x − 122.28 (r² = 0.99, n = 4, P < 0.05).It was concluded that dietary lysine level had a significant influence on NR and whole-body protein turnover but not on plasma IGF-I and GH concentration. Plasma IGF-I may be an important factor controlling N metabolism of growing pigs. Further research was needed to study the mechanism.