Expression of Frizzled 2 in the mouse ovary during oestrous cycle

The Wnt/β‐catenin pathway is a conserved signalling pathway that regulates gene expression and controls diverse developmental processes such as cell fate specification, cell proliferation and cell differentiation. To our knowledge, the potential role of this pathway in the adult ovary has been poorly addressed. To this end, we investigated the expression pattern of Frizzled 2 in the mouse ovary during oestrous cycle by real‐time Q‐PCR, in situ hybridization, Western blotting and immunohistochemistry. In this study, we used uterine wet weight and H3.2 mRNA level as two markers for classification of mouse oestrous cycle. We found that both uterine wet weight and H3.2 mRNA are the highest in the proestrus and decrease from oestrus to diestrus. During oestrous cycle, Frizzled 2 mRNA and protein exhibited the highest level in the proestrus stage and rapidly reduced from oestrus to diestrus. In situ hybridization results showed that the positive signals for Frizzled 2 are highly detected in the oocyte and stroma in proestrus stage, while moderate or weak Frizzled 2 mRNAs are localized in the oocyte, granulosa cells, stroma and corpus luteum from oestrus to diestrus. The localization pattern of Frizzled 2 protein is mostly consistent with its mRNA, but stronger Frizzled 2 proteins are present in the granulosa cells and membrane of oocyte in oestrous cycle. Our data suggest that Frizzled 2 may be involved in regulating follicle growth, oocyte maturation and luteinization during oestrous cycle.     

Endometrial gene expression in primiparous dairy cows at the end of the voluntary waiting period is affected by nutrition: Total mixed ration vs increasing levels of herbage allowance

The study postulated that differential nutritional management during the early lactation period would be reflected in endometrial expression of genes related to embryo growth at the end of the voluntary waiting period. Thus, the effect of the combined use of total mixed ration (TMR) and grazing under different herbage allowances during the first 75 days post‐partum (DPP) on endometrial gene expression was evaluated in primiparous dairy cows. Cows were blocked by body weight, age and body condition score and randomly assigned to three grazing treatments: high (HA, 30 kg DM per cow per day), medium (MA, 15 kg DM per cow per day) and low (LA, 7.5 kg DM per cow per day) herbage allowance (mixed pasture, 2,600 kg DM per ha) plus 8 kg DM of supplement or TMR (55% forage, 45% concentrate) fed ad libitum (TMR) from calving to 75 DPP. At 57 DPP, cows were synchronized for oestrus (day 0, 68 DPP) and at day 7, endometrial biopsies were obtained. The nutritional treatment did not affect insulin, IGF‐1 and leptin concentrations on days 0, 4 or 7. Expression of IGF1, IGFBP3, IGFBP4, ADIPOR1 and ADIPOR2 mRNA was significantly affected by the nutritional treatment. Endometrial IGF1 and IGFBP4 mRNA were twofold greater in TMR and HA than MA and LA cows. Expression of IGFBP3 and ADIPOR1 mRNAs was greater in TMR and HA than MA cows, but did not differ from LA cows. All groups had greater expression of ADIPOR2 mRNA than MA cows. This study provided solid evidence of the importance of nutritional management during early lactation on uterine environment at the end of the voluntary waiting period. The greater expression of genes related to embryo growth and uterine function (IGF system, progesterone and adiponectin receptors) in cows fed diets maximizing energy intake suggests a favourable environment for embryonic growth, which may explain the improved reproductive performance of cows in good energy balance.     

The post-ovulatory rise in progesterone is lower and the persistence of oestrous behaviour longer during the first compared with the second cycle of the breeding season in mares

Mares are seasonally polyoestrous breeders. Therefore, the first ovulation of the season, following winter anoestrus, is the only cycle in which mares ovulate without the presence of an old CL from the previous cycle. The objective of this study was to compare the length of oestrous behaviour, and plasma progesterone concentrations during the early post-ovulatory period between mares after the first and second ovulation of the breeding season. Overall, 38 mares and 167 oestrous periods were used in the study. From those, 11 mares were used during the first and subsequent oestrous period to measure and compare the post-ovulatory rise in progesterone concentration, whereas all the mares were used to compare the length of the post-ovulatory oestrous behaviour between the first and subsequent cycles of the breeding season. The persistence of the post-ovulatory oestrus was longer (p < .001) following the first ovulation of the year (median of 52 h) compared with the subsequent ovulations (median of 36 h for second and later ovulations groups; n = 38 mares). The progesterone concentration at any of the four 8 h-intervals analysed (28, 36, 76 and 84 h post-ovulation) was lower (p < .01) following the first versus the second ovulation of the year. By 36 h post-ovulation the progesterone concentration of mares at the second ovulation of the year had passed the threshold of 2 ng/ml (2.1 ± 0.33 ng/ml), whereas in the first cycle it was 1.2 ± 0.13 ng/ml. In conclusion, mares had lower progesterone concentrations in their peripheral circulation and longer persistence of oestrous behaviour following the first ovulation of the year compared with the second and subsequent ovulatory periods of the breeding season.     

Seminiferous epithelium cycle staging based on the development of the acrosome in ram testis

Testicular histopathology is considered the most sensitive and reliable method to detect the effects of chemicals on sperm production. To carry out a sensitive examination of testicular histopathology and interpret the changes require knowledge of spermatogenic stages. Spermatogenic staging based on acrosome development during spermiogenesis is conventionally performed in animal species routinely used for research and toxicity testing. In contrast, small ruminants, such as sheep and goats, are rarely used as animal models to evaluate toxicity in male reproductive organs. To the best of our knowledge, a comparable spermatogenic staging system in rams has not yet been fully characterised. Hence, this study aimed to adapt the existing spermatogenic staging based on acrosome development in bull testes to fit the seminiferous epithelium cycle of ram testes. The results show that spermatogenic staging based on acrosome development in bull testes can, with slight modifications, be efficiently used for the staging of ram testes.     

Results of vaginal cytology studies and examination of the arborization phenomenon of the vaginal mucus for the determination of estrus cycle activity in sheep]

Vaginal smears were taken by means of a metal spatula from the dorsal part of the vaginal vestibule of sexually mature sheep with intach genitals for the purpose of following up changes in the vaginal cell structure. The smears were taken daily over the whole testing period of 35 days. The cells were defferentiated by means of the dyeing method according to Papanicolaou as well as by using Shorr's staining which had been modified according to Pundel. Differentiation was made between occurrences ob basal, parabasal, intermediary, and superficial cells, with the view to figuring out the cycle phases. That vaginal smear method, when used in two or three folloow-up examinations on the basis of random checks, has proved to enable determination of cyclic activity of a complete herd. This is of great importance to heart synchronization or heart induction, since all results so far have shown that all success depended strongly on the given degree of cyclic activity of the sheep concerned.     

Development of enzyme immunoassay for chromogranin A (CgA) and profiling of plasma CgA concentrations in the cow

The objectives of this study were to establish and characterize a homologous immunoassay for bovine chromogranin A (bCgA) and to profile plasma bCgA concentrations during early pregnancies. We synthesized oligopeptide corresponding to the amino acid sequence 341–355 of bCgA for immunizing rabbits and peptide corresponding to the amino acid sequence 336–365 of bCgA for both a biotinylated tracer and reference standards. Recombinant bCgA protein was also generated in Escherichia coli lysate. Dose-dependent displacement curves were obtained from 1 to 1,000 nM of the reference standards. The displacement curves showed a good relationship between the reference standards of the synthetic peptide and the serially diluted plasma sample or recombinant bCgA protein generated in the present study. The assay sensitivity defined as the value of two standard deviations below the zero standard was calculated as 0.46 nM. The intraassay and interassay coefficients of variation were 6.48% and 13.4%, respectively. Changes in the plasma bCgA concentrations in early pregnancies undulated in nonpregnant animals. The results of the present study suggest that assaying plasma bCgA concentrations could be utilized as measures to evaluate the physiological status of cattle.     

日粮精粗比对奶牛瘤胃内环境的影响

本文主要讨论了日粮精粗比对奶牛瘤胃内环境的影响,包括对瘤胃细菌菌群数量、瘤胃pH值、瘤胃内容物稀释率和外流速度、氨态氮浓度以及挥发性脂肪酸的影响。     

Evaluating the suitability of field beans as a substitute for soybean meal in early-lactating dairy cow: Production and metabolic responses

Twenty-four multiparous Holstein cows were utilized in a completely randomized design to examine the effect of feeding field beans (FB, Vicia faba L. var. minor) as substitute for soybean meal (SBM) on lactation performance and metabolic response during the early lactation period. Cows were individually divided into two equal groups and fed for 16 weeks on one each of the two experimental diets. The controls were fed pelleted concentrate contained 150 g/kg dry matter (DM) of SBM as the main protein source, whereas the experimental concentrate contained 345 g/kg DM of FB. Oat hay was offered ad libitum to cows and water was freely available. Blood samples were assayed for their content of: urea, glucose, triglycerides, cholesterol, total protein, albumin, calcium and phosphorus, aspartate aminotransferase (AST) and alanine aminotransferase (ALT), nonesterified fatty acids (NEFA) and β-hydroxybutyrate (BHBA). Inclusion of FB had no detectable effects on DM intake (DMI), body weight (BW), or body condition score (BCS). Neither milk yield nor quality were influenced by dietary treatment, except for milk urea nitrogen (MU) that was reduced in cows fed the FB diet (P < 0.05). Clotting properties of milk were not affected adversely by added dietary FB. Concentration of blood urea (BU) was lower in cows fed the FB diet than in those fed the control SBM diet (P < 0.05). These findings indicate that feeding FB in a lactation diet supported lactation performance similar to cows fed traditional SBM-based diet, and the results may elicit great interest for countries where soybean utilization is adversely influenced by high supply costs.     

乳牛衣原体病灭活疫苗效检小鼠模型的建立

为了解决以本动物效检乳牛衣原体病灭活疫苗成本高的问题，用11批乳牛衣原体病灭活疫苗做乳牛效力试验时同步做小鼠效力试验，除第3批次疫苗的本动物效检结果与小鼠效检结果略有差异外，其他批次疫苗的效检结果均一致。结果表明，用小鼠做牛衣原体灭活疫苗的效检动物模型是可行的。     

Comparison of different models for estimation of genetic parameters of early growth traits in the Mehraban sheep.

Genetic parameters for birth weight (BW), weaning weight (WW) and pre-weaning daily gain (PWDG) in Iranian Mehraban sheep were estimated using restricted maximum likelihood (REML) procedure. Six different animal models were fitted, differentiated by including or excluding maternal effects, with and without covariance between maternal and direct genetic effects. The estimates for direct heritability ranged from 0.26 to 0.53, 0.18 to 0.32 and 0.15 to 0.33 for BW, WW and PWDG respectively. The estimates were substantially higher when maternal effects, either genetic or environmental, were ignored in the model. The results of this study show that full models with maternal genetic and environmental effects gave the most accurate estimates for early growth traits.     

利用体外培养法建立饲料营养成分与瘤胃丙酸产量的回归模型

根据美国康乃尔净碳水化合物和蛋白质体系 (CNCPS) ,分析测定了 4类 2 4种饲料的营养成分 ,利用体外批次培养技术测定了它们在体外培养一定时间后的丙酸产量 ,并由此建立了饲料中产丙酸的营养成分和瘤胃丙酸产量的回归方程     

Use of heterogeneous operation-specific contact parameters changes predictions for foot-and-mouth disease outbreaks in complex simulation models

The role of contact parameters in a complex spatial simulation model of foot-and-mouth disease spread was determined by comparing predictions of number of infected premises, epidemic duration, and relative infection risk for different production sectors between a model that included the Full, heterogeneous (differing by production type) type-specific information about animal, vehicle and personnel movement between premises, and models that used partial and homogeneous (similar across production types) weighted-mean or proxy parameter sets for contacts between premises of all types. The model was run using a dataset of known premises locations in a three-county area in the Central Valley of California and categorized into 13 premises types and six production sectors.Results from models run with homogeneous contact parameters were always different from those obtained from the Full model, demonstrating that model predictions are affected by heterogeneity in contact parameters. Models simplified by using weighted-mean parameters predicted fewer infected premises. Models that were simplified by using medium dairy farm or large swine operation proxy parameters predicted longer epidemics with more infected premises, while those using small beef operation proxy parameters predicted shorter epidemics with fewer infected premises. Simplified-parameter models underestimated the impact on the economically important dairy sector, while overestimating the impact on beef and backyard operations. Results establish a need for heterogeneous, operation-specific contact parameters in complex stochastic simulation models that must be weighed against the cost of obtaining and coding premises type-specific contact information.     

Conformity and controversies in the diagnosis,staging and follow‐up evaluation of canine nodal lymphoma: a systematic review of the last 15 years of published literature

Diagnostic methods used in the initial and post‐treatment evaluation of canine lymphoma are heterogeneous and can vary within countries and institutions. Accurate reporting of clinical stage and response assessment is crucial in determining the treatment efficacy and predicting prognosis. This study comprises a systematic review of all available canine multicentric lymphoma studies published over 15 years. Data concerning diagnosis, clinical stage evaluation and response assessment procedures were extracted and compared. Sixty‐three studies met the eligibility criteria. Fifty‐five (87.3%) studies were non‐randomized prospective or retrospective studies. The survey results also expose variations in diagnostic criteria and treatment response assessment in canine multicentric lymphoma. Variations in staging procedures performed and recorded led to an unquantifiable heterogeneity among patients in and between studies, making it difficult to compare treatment efficacies. Awareness of this inconsistency of procedure and reporting may help in the design of future clinical trials.     

A Review of Simulation Modelling Approaches Used for the Spread of Zoonotic Influenza Viruses in Animal and Human Populations

Increasing incidences of emerging and re‐emerging diseases that are mostly zoonotic (e.g. severe acute respiratory syndrome, avian influenza H5N1, pandemic influenza) has led to the need for a multidisciplinary approach to tackling these threats to public and animal health. Accordingly, a global movement of ‘One‐Health/One‐Medicine’ has been launched to foster collaborative efforts amongst animal and human health officials and researchers to address these problems. Historical evidence points to the fact that pandemics caused by influenza A viruses remain a major zoonotic threat to mankind. Recently, a range of mathematical and computer simulation modelling methods and tools have increasingly been applied to improve our understanding of disease transmission dynamics, contingency planning and to support policy decisions on disease outbreak management. This review provides an overview of methods, approaches and software used for modelling the spread of zoonotic influenza viruses in animals and humans, particularly those related to the animal‐human interface. Modelling parameters used in these studies are summarized to provide references for future work. This review highlights the limited application of modelling research to influenza in animals and at the animal‐human interface, in marked contrast to the large volume of its research in human populations. Although swine are widely recognized as a potential host for generating novel influenza viruses, and that some of these viruses, including pandemic influenza A/H1N1 2009, have been shown to be readily transmissible between humans and swine, only one study was found related to the modelling of influenza spread at the swine‐human interface. Significant gaps in the knowledge of frequency of novel viral strains evolution in pigs, farm‐level natural history of influenza infection, incidences of influenza transmission between farms and between swine and humans are clearly evident. Therefore, there is a need to direct additional research to the study of influenza transmission dynamics in animals and at the animal‐human interface.     

雌二醇与孕酮奶样纸片酶免法建立及其临床应用

乳汁雌二醇与孕酮含量是判断母牛卵巢机能状态的重要指标。本试验建立了雌二醇与孕酮奶样纸片酶免法,并用于测定不同定时排卵程序中母牛乳汁雌二醇与孕酮含量,结果表明所建立的方法简便实用,排卵程序中母牛乳汁孕酮、雌二醇含量变化规律与理论分析相符,可以用来判定母牛发情、妊娠及卵巢功能。     

Multi-trait animal model estimation of genetic parameters for linear type and gait traits in the Belgian warmblood horse

Genetic parameters for the height at withers, 27 linear type and six linear gait traits were estimated for the Belgian warmblood horse. Observations on 987 mares, mostly 3 years old, were analysed using a multi-trait animal model. The statistical model included appraiser, age and location (date × place of appraisal) as fixed effects. Genetic parameters were estimated using a canonical transformation and an expectation-maximization restricted maximum likelihood algorithm with an additional deceleration step. Estimates of heritability for the 33 linear traits were between 0.15 and 0.55. Heritability of the height at withers was 0.34 ± 0.06. Estimated genetic correlations ranged from −0.60 to 0.98 with an average SE of 0.10. The highest positive correlations were found among traits of walk and among traits of trot. Volume and the quality of legs were the most negatively correlated. Estimated genetic parameters indicated that the linear scoring system is a valuable tool to assess conformation. The full (co)variance matrix is now available for breeding value estimation to support selection for conformation and gaits.     

Estimation of genetic parameters for longevity traits in dairy cattle: A review with focus on the characteristics of analytical models

Longevity is an economically important trait of dairy cattle for increasing the profitability of dairy management. The reasons for culling can be either voluntary (primarily productivity) or involuntary (primarily health and fertility). Longevity characteristics include: (i) true longevity (all culling reasons, including productivity); and (ii) functional longevity (all culling reasons, except productivity). Improvements to longevity are made to decrease the rate of involuntary culling rather than extend the herd life (HL). The proportional hazard model is useful for evaluating genetic ability for HL. However, the differences between estimates made using the proportional hazard model and those made using linear single or multiple‐trait animal models are not clear. The model commonly used for evaluation differs among countries. Productive traits, udder traits, and feet and legs traits are genetically correlated with longevity, and consequently these traits are used to indirectly evaluate longevity. The reliability of estimates of genetic ability for longevity is increased by combining direct and indirect estimates. In Japan, HL is evaluated using the multiple‐traits model. The genetic correlations between HL and other traits vary with the birth year. Therefore, these genetic correlations need to be reviewed regularly.     

Development of a model for inducing transient insulin resistance in the mare: preliminary implications regarding the estrous cycle

Peripheral insulin resistance is the failure of proper cellular glucose uptake in response to insulin. Insulin resistance and hyperinsulinemia are associated with several disease states in the horse and reproductive function disturbances in humans, including polycystic ovarian syndrome. To test the hypothesis that insulin resistance (IR) and hyperinsulinemia disrupt the estrous cycle in mares, two experiments were conducted to first develop a model to induce IR and to then examine the effect of this model on the duration of the estrous cycle. In Exp. 1, a hyperinsulinemic-euglycemic clamp (HEC) procedure was performed on seven mares to determine insulin sensitivity before and immediately following infusion of a heparinized lipid solution. The HEC procedure was repeated 1 wk after lipid infusion. Mares developed IR following the lipid infusion (P < 0.05), and some individuals maintained IR for up to 1 wk. Mares also exhibited increased blood insulin both immediately following treatment and 1 wk later (P < 0.05). In Exp. 2, induction of insulin resistance by lipid solution was not accompanied by changes in circulating concentrations of luteinizing hormone, and duration of the luteal phase, compared with the duration of untreated luteal phases. Nonetheless, lipid infusion and the resultant insulin resistance were associated with an increased interovulatory period (P < 0.05), and peak concentrations of progesterone (P < 0.05) were higher during the treated vs. untreated luteal phases of the estrous cycle. The results from the preliminary study suggest that infusion of a lipid solution may induce transient insulin resistance and hyperinsulinemia. The resulting insulin resistance and hyperinsulinemia may modify characteristics of the estrous cycle, perhaps at the level of the ovary.     

The utilisation of a high level energy/protein diet for lactating Mediterranean buffaloes: Intake capacity and effects on quanti–qualitative milk parameters

The trial was carried out using 14 multiparous buffaloes divided into two groups according to yield in the previous lactation (2653 vs. 2625 kg), number of lactations (4.8 vs. 4.1), live weight (670.6 vs. 657.1 kg), days post-calving (21.6 vs. 23.6 days) and average yield at the on-set of the trial (11.18 vs. 10.90 kg/day). The effects of both feed intake and the quanti–qualitative parameters of the milk yield were evaluated following the ad libitum administration of two diets containing two different forage/concentrate ratios, diet 1 = 48 : 52 (0.94 Milk FU/kg DM, 158.3 g/kg DM of crude protein); diet 2 = 55 : 45 (0.85 Milk FU/kg DM, 144.4 g/kg DM of crude protein). The trial lasted for 114 days, on average from day 23 to 137 of lactation. The dry matter intake for diet 1 was 16.37 kg/day, for diet 2 it was 16.53 kg/day. No significant differences were found with live weight of the animals; however, the buffaloes in group 1 showed gains of 35.0 kg compared to 54.2 kg of those in group 2. A significant difference (P < 0.05) in milk yield favouring group 1 (11.66 vs. 9.42 kg/day) was observed while fat (73.20 vs. 80.08 g/kg) and protein content (43.24 vs. 46.41 g/kg) was higher for group 2. Coagulation data indicated that the milk yield of group 2 showed better coagulation properties (r = 18.35 min, K₂₀ = 3.88 min, A₃₀ = 47.00 mm), than group 1.