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A major limitation to producing high seed yields in berseem clover (Trifolium alexandrinum L.) is failure to set seed, predominantly due to lack of pollination. Despite the importance of berseem clover as a leading forage legume, the contribution of pollinators to seed set is scarce. In Pakistan, the honeybee population is declining mainly because of the extensive use of neonicotinoid pesticides and habitat fragmentation. This, combined with the region's harsh environment and the use of inferior, locally bred genotypes, has resulted in low seed yields. Insufficient seed availability leads to limited forage supply, resulting in poor livestock nutrition, which subsequently impacts livestock health and productivity, and reduces farmers' income. The present study estimated the seed production of 3 berseem clover genotypes resulting from honeybees [Apis mellifera L. (Hymenoptera: Apidae)] pollination in 2 growing seasons (2012-2014) in the central Punjab region of Pakistan. Experiments had 2 pollination treatments (open pollination and honeybee inclusion) and 3 seed genotypes, viz. farmer own-saved, market, and the improved variety cultivars. For both growing seasons, honeybee pollination resulted in significant increases in seed yields ranging from 35% to 67%, regardless of seed genotype. With the exception of the number of seed heads/m2, all seed yield parameters also increased significantly in response to honeybee inclusion. The combination of improved variety and honeybee inclusion resulted in the production of a maximum number of seeds per head (45.3), 1,000-seed weight (3.7 g), and estimated seed yield (375.5 kg/ha). In addition, the increase in estimated net income of seed ranged from PKR 82,485 Rs/ha (US$844/ha) to PKR 168,975 Rs/ha (US$1728/ha) with the use of honeybees as an insect pollinator across all the seed genotypes. Honeybee pollination has broader implications for mixed farming systems by playing a key role in preserving biodiversity and promoting sustainable agriculture. It also enhances the quality and quantity of berseem crops by increasing the production of high-quality seeds and forage leading to improved livestock productivity and family food security which strengthens the economic resilience of rural communities., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America.)

Global health programs engaging in isolated or short-term medical missions can and do cause harm, reinforce health care disparities, and impede medical care in the regions where it is so desperately needed. Related ethical, medical, and legal concerns are reviewed in this article. The authors recommend abandoning these ill-considered missions and focusing attention and resources on advancing neurology through ethically congruent, multisectoral, collaborative partnerships to establish sustainable, self-sufficient training programs within low- and middle-income countries., (Copyright © 2023 Elsevier Inc. All rights reserved.)

This article provides an overview of current malpractice trends in neurology as well as non-malpractice and forensic liability concerns. It is more important for clinicians to recognize the common patient care scenarios that are likely to precipitate lawsuits rather than memorize arcane legal principles. Therefore, this article offers an introduction to malpractice jurisprudence as well as a general overview of current litigation trends and a review of the role and duties of a neurologist serving as an expert witness. The next article highlights mitigation strategies for the most prevalent neurologic misadventures., (Copyright © 2023 Elsevier Inc. All rights reserved.)

This chapter highlights the most frequently encountered neurological malpractice claims. The format is designed to provide a rudimentary understanding of how lawsuits arise and thereby focus discussion on adapting practice patterns to improve patient care and minimize liability risk., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Sialic acids (Sias) are acidic monosaccharides and red meat is a notable dietary source of Sia for humans. Among the Sias, N-acetylneuraminic acid (Neu5Ac) and 2-keto-3-deoxy-D-glycero-D-galacto-2-nonulosonic acid (KDN) play multiple roles in immunity and brain cognition. On the other hand, N-glycolylneuraminic acid (Neu5Gc) is a non-human Sia capable of potentiating cancer and inflammation in the human body. However, their expression within the animal kingdom remains unknown. We determined Neu5Ac and KDN in skeletal muscle and organs across a range (n = 9) of species using UHPLC and found that (1) caprine skeletal muscle expressed the highest Neu5Ac (661.82 ± 187.96 µg/g protein) following by sheep, pig, dog, deer, cat, horse, kangaroo and cattle; (2) Among organs, kidney contained the most Neu5Ac (1992−3050 µg/g protein) across species; (3) ~75−98% of total Neu5Ac was conjugated, except for in dog and cat muscle (54−58%); (4) <1% of total Sia was KDN, in which ~60−100% was unconjugated, with the exception of sheep liver and goat muscle (~12−25%); (5) Neu5Ac was the major Sia in almost all tested organs. This study guides consumers to the safest red meat relating to Neu5Ac and Neu5Gc content, though the dog and cat meat are not conventional red meat globally.

Chitosan oligosaccharide (COS) is derived through deacetylation of chitin from crustacean shells. Previous studies reported the benefits of COS to gut microbiota, immunity and health of host species. In this study, 120 pregnant composite ewes were subdivided into treatment and control groups in duplicate. COS was supplemented via a loose lick to provide an estimated intake of COS @100−600 mg/d/ewe for five weeks pre-lambing until lamb marking. Body weight was recorded pre-treatment for ewes, and at lamb marking and weaning for both ewes and lambs. Serum immunity markers immunoglobulin G (IgG), immunoglobulin M (IgM), immunoglobulin A (IgA), secretory immunoglobulin A (sIgA), interleukin (IL)-2, IL10 and faecal sIgA were determined for ewes and lambs at lamb marking and weaning by enzyme-linked immunosorbent assay (ELISA). We found that COS can be incorporated in sheep feed without compromising palatability. Maternal COS supplementation did not influence the body weight of ewes or lambs. It did, however, significantly increase the concentrations of serum IL2 in ewes at marking and weaning (p < 0.001). In lambs, COS also significantly increased the IL2 concentration at making (p = 0.018) and weaning (p = 0.029) and serum IgM at marking (p < 0.001). No significant effect was observed in the concentration of any other immune marker or cytokine in either ewes or lambs. In conclusion, maternal COS supplementation significantly modulated some immunity markers in both ewes and lambs. The short duration of maternal COS supplementation and optimal seasonal conditions during the trial may explain the lack of significant body weight in ewes and lambs from the COS supplementation.

Numerous catastrophic events in the 21st century have motivated renewed discussion regarding whether the traditional definition of standard of care appropriately applies to clinical decision-making in crisis scenarios. Some authorities have proposed the adoption of a crisis standard of care, which refines physician responsibilities during a crisis event in accordance with population health principles. However, this proposal is fraught with controversy, and current medical and legal scholarship on this topic remains complex and conflicted. To clarify these points and provide practicing neurosurgeons with guidance, we provide a review of current literature on the evolving definitions of crisis standard of care. Additionally, we provide an assessment of the implications of a crisis standard of care, as it relates to legal liability, clinical ethics, and neurosurgical practice., (Copyright © Congress of Neurological Surgeons 2022. All rights reserved.)

This study aimed to evaluate the sensory and physical characteristics of zingibain-injected meat combined with sous vide cooking. M. biceps femoris (BF; n = 12) acquired from 6-7 year old Angus cows were cooked using the sous vide method at 65 °C, for 8 h or 12 h, either with ginger powder (GP) injected in a 2 g/L solution in water (treatment) or un-injected (control). The sensory attributes included flavour, juiciness, tenderness, and physicochemical characteristics were Warner-Bratzler shear (WBSF), hardness, total water content (TWC), cooking loss (CL) and collagen content. A significant improvement in tenderness with injection treatment and cooking time was observed, as evaluated through trained sensory panellists, and reduced WBSF and hardness ( p < 0.05 for all). The flavour of the meat was not affected by injection treatment or cooking time ( p > 0.05), but juiciness and TWC were reduced with longer cooking times ( p < 0.01 for both). Soluble collagen increased with injection treatment and cooking time (both p < 0.05). Moderate to high correlations were found between sensory and physical measurements for tenderness and juiciness. The longer cooking time (12 h) with GP injection treatment caused over tenderization of the meat. The soft texture associated with over-tenderization may be suitable for some specialised consumer markets, for instance, the elderly population with chewing difficulties. Improving the eating quality of low-quality meat from old animals through sous vide cooking and the use of ginger proteases may increase the acceptability of lower value beef, potentially enhancing the commercial value of carcasses typically produced in the beef industry.

Red meat-derived sialic acid (Sia), N-glycolylneuraminic acid (Neu5Gc), promotes the risk of carcinoma and inflammation. Expression in skeletal muscle and organs across animal species remains unknown. We measured Neu5Gc in skeletal muscle and organ tissues from nine species using UHPLC and found that: (1) neu5Gc concentration in skeletal muscle was highest in goats (166 ± 48.7 µg/g protein), followed by cattle, pig, sheep, horse, cat and deer: >75% was conjugated. No Neu5Gc was detected in kangaroo and dog muscles; (2) total Neu5Gc in organ meats was generally about 2-54% higher than in muscle. Surprisingly Neu5Gc was absent in seven organs of female deer; (3) nine commercial ovine meat cuts contained similar Neu5Gc levels. Thus, red meat Neu5Gc concentration is tissue and species-specific and absent in muscle and organ tissue of some species. Our study provides guidelines for animal meat preferences for consumers and sheds light on the functionality of Neu5Gc., (Crown Copyright © 2020. Published by Elsevier Ltd. All rights reserved.)

Chitosan oligosaccharides (COS) are the hydrolyzed product of chitosan and have multifunctional health benefits. The objective of this study was to elucidate the effect of COS as a dietary supplement to gilts on their productivity and health and that of their litters. Gilts were randomly assigned to either a treatment ( n =  30) or control group ( n =  30). The treatment gilts were fed a standard dry sow ration supplemented with COS at 0.12 and 0.24 g/gilt per d during gestation and lactation, respectively, and the control group was fed the standard dry sow ration only. The body weight, reproductive performance, milk production and litter size for each gilt and body weight of corresponding litters were recorded. The serum immunoglobulins (IgA, IgG, IgM) and secretory immunoglobulin A (sIgA) concentrations of gilts and piglets and fecal sIgA concertation of gilts were measured by Enzyme-linked immunosorbent assay (ELISA). Our study showed that maternal COS supplementation 1) significantly increased gilt body weight in late pregnancy (P <  0.05), 2) significantly increased milk production of gilts at different stages (d 1, 3, 7 and 19) of lactation (P <  0.05), 3) significantly increased body weight gain of piglets at weaning ( P <  0.05), 4) significantly increased the serum concentrations of IgM and sIgA in piglets, and sIgA in fecal sample of gilts (P <  0.05), and 5) tended to increase the pregnancy success rate (P >  0.05) in the treatment group compared to the control group. These results suggest that maternal COS intervention in gilts can improve gilt milk production, piglet pre-weaning growth and immunity parameters in both gilts and piglets., Competing Interests: We declare that we have no financial and personal relationships with other people or organizations that can inappropriately influence our work, there is no professional or other personal interest of any nature or kind in any product, service and/or company that could be construed as influencing the content of this paper., (© 2020 Chinese Association of Animal Science and Veterinary Medicine. Production and hosting by Elsevier B.V. on behalf of KeAi Communications Co., Ltd.)

A field study was conducted on smallholder farmer fields between 2012 to 2014 to evaluate the performance of cv. Agaitti Berseem-2002, against local landraces exchanged between farmers (LBF1) or available from local markets (LBM1). The effects of genotype and harvesting regimen on forage production, quality and seed production were evaluated. Significant differences (P < 0.05) among genotypes and cutting treatments were recorded for forage and seed yields, and forage quality across all research sites in both years. Maximum cumulative fresh forage (89.7 t/ha) and dry matter (DM; 13.4 t/ha) yields were obtained with Agaitti Berseem-2002 when harvesting occurred five times over the season. However, maximum seed yield (1048 kg/ha) with higher 1000-seed weight (3.63 g) were obtained if forage was only harvested three times and the crop then left for seed set. Agaitti Berseem-2002 also produced forage with the higher crude protein content (27%), DM digestibility (69%), digestible organic matter (DM basis; 65%) and metabolizable energy content (10%) compared to the local landraces (LBF1 and LBM1). Therefore, the harvesting regimen for greatest economic return which produced optimum fresh and DM forage yields of highest nutritive values and maximum seed yield, were comprised of taking three forage cuts (at 65, 110 and 150 days after sowing) prior to seed harvest.

Milk oligosaccharides (MOs) are complex carbohydrates with multifunctional health benefits for the neonate. Poor reproductive performance in primiparous gilts limits their productivity. Changes in the structure and abundance of porcine MO (PMOs) through lactation with parity remains unknown and may explain superior new-born growth in litters from multiparous sows relative to gilts. We report 55 PMOs structures, of which 25 are new (17 sialylated and 8 neutral). Their incidence in gilt and sow colostrum was almost identical (53 vs. 54), but not in transitional milk (48 vs. 53) nor mature milk (41 vs. 47). These PMOs including neutral-, sialyl- and fucosyl- MOs in colostrum were more abundant in the gilt than the sow, but always decreased during lactation. Structural diversity decreased, although fucosylated MO were conserved. In conclusion, high diversity and levels of MO in porcine milk is parity dependent. Given the similarity between porcine and human MO profiles, our findings may help define key roles for MOs as potential dietary additives to improve growth of neonates from first pregnancies in both human and sows.

Lactoferrin (LF), a sialylated iron-binding glycoprotein, performs multiple beneficial functions including modulating immunity and improves neurodevelopment, health and growth performance. Maternal LF intervention for gilts (first parity sows) on the performance of gilts and their offspring remains unknown. In the current study gilts were fed with a commercial pig feed supplemented with 1g LF /day (treatment group) or 1g milk casein/day (control group) from day 1 post mating throughout pregnancy and lactation for about 135 days. The milk production and body weight gain was monitored. The immunoglobulin concentrations in the serum of gilts and piglets were measured using ELISA. Our study showed that maternal LF supplementation to the gilt (1) significantly increased milk production at different time points (day 1, 3, 7 and 19) of lactation compared to the control (p<0.001); (2) significantly increased body weight gain of their piglets during the first 19 days of life compared to the control group (p<0.05); (3) tended to increase pregnancy rate, litter size and birth weight, number of piglets born alive, and decrease the number of dead and intrauterine growth restriction (IUGR) piglets; (4) significantly increased the concentration of serum IgA in gilt and serum sIgA in piglet (p<0.05). In summary, maternal Lf intervention in gilts can improve milk production, pig production and serum IgA and sIgA levels, and therefore plays a key role in shaping the performance of their progeny.

Managing the return to regular cyclicity after the winter anestrous period in the mare has been a challenge for the equine breeding industry. Specifically, efforts have been made to shift or shorten the vernal transition period and to have it followed by a predictable first ovulation at the commencement of the breeding season. Intravenous administration of kisspeptin is known to stimulate an LH response in both reproductively active and inactive mares. This study examined the effects of a constant rate infusion (CRI) of kisspeptin on mares during vernal transition. Mares were given a 30 hours infusion of kisspeptin at a low and high rate (66 nmol [88 μg] and 100 nmol [130 μg] per hour, respectively) or saline, and the LH and follicular response tracked. Plasma samples were collected every 15 minutes for the first 6 hours to determine if there is an acute effect of kisspeptin infusion on LH secretion. Plasma samples were then collected every 3 hours for a total of 72 hours to examine the ability of kisspeptin to stimulate an LH surge. A CRI of kisspeptin increased LH secretion in these mares but was not able to stimulate an LH surge. To examine the effect of kisspeptin on ovarian activity, follicular measurements were collected ultrasonographically until ovulation occurred or the follicles regressed. CRI of kisspeptin at these rates was unable to induce ovulation earlier than controls., (Copyright © 2016 Elsevier Inc. All rights reserved.)

Sialic acids (Sia) are key monosaccharide constituents of sialylated glycoproteins (Sia-GP), human sialylated milk oligosaccharide (Sia-MOS), and gangliosides. Human milk sialylated glycoconjugates (Sia-GC) are bioactive compounds known to act as prebiotics and promote neurodevelopment, immune function, and gut maturation in newborns. Only limited data are available on the Sia content of porcine milk. The objective of this study was to quantitatively determine the total level of Sia N-acetylneuraminic acid (Neu5Ac), N-glycolylneuraminic acid (Neu5Gc), and ketodeoxynonulosonic acid (KDN) in porcine milk and to compare these levels in gilt and sow milk during lactation. Milk from 8 gilts and 22 sows was collected at 3 stages of lactation (colostrum, transition, and mature milk). Standard and experimental samples were derivatized using 1,2-diamino-4,5-methylenedioxy-benzene and analyzed by ultra-high-performance liquid chromatography using a fluorescence detector. The following new findings are reported: (1) Gilt and sow milk contained significant levels of total Sia, with the highest concentration in colostrum (1,238.5 mg/L), followed by transition milk (778.3 mg/L) and mature milk (347.2 mg/L); (2) during lactation, the majority of Sia was conjugated to Sia-GP (41-46%), followed by Sia-MOS (31-42%) and a smaller proportion in gangliosides (12-28%); (3) Neu5Ac was the major form of Sia (93-96%), followed by Neu5Gc (3-6%) and then KDN (1-2%), irrespective of milk fraction or stage of lactation; (4) the concentration of Sia in Sia-GP and Sia-MOS showed a significant decline during lactation, but the level of ganglioside Sia remained relatively constant; (5) mature gilt milk contained a significantly higher concentration of Sia-GP than sow milk. The high concentration of total Sia in porcine milk suggests that Sia-GC are important nutrients that contribute to the optimization of neurodevelopment, immune function, and growth and development in piglets. These findings provide an important rationale for the inclusion of Sia-GC in pig milk replacers to mimic porcine milk composition for the optimal growth and development of piglets., (Copyright © 2016 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

The aim of this study was to identify parameters that may be used as objective measures of wellbeing in weaner pigs on farm. Eighty-six weaner pigs were categorized into 3 groups based on their BCS: very compromised (VC, BCS = 1), moderately compromised (MC, BCS = 2), or healthy (HY, BCS ≥ 3). Nonphysiological and physiological measures were then collected from the pigs. Nonphysiological measures included 3 subjective binary scores that assessed the activity, hairiness, and responsiveness of each pig. Physiological measures included rectal temperature, skin temperature, and 11 blood-based parameters. Each physiological measure was assessed in a ternary manner, with pigs being classed as being above, below, or within the normal physiological range. Fisher’s exact tests (FET) were used to identify differences between the numbers of pigs in each BCS category for each variable, and ANOVA of physiological measures was also conducted. Correlations between all variables were also conducted. The FET results identified significant differences for 8 of the measures, with activity, hairiness, responsiveness, rectal temperature, and blood concentrations of glucose, creatinine kinase (CK), albumin, and globulin all differing significantly (all P < 0.05) according to category. Rectal temperature, glucose, and globulin concentrations were significantly different in ANOVA analyses. Of these measures, there were significant correlations between hairiness, rectal temperature, and glucose (all R > 0.30 and P < 0.05). Based on these results, rectal temperature, glucose, and hairiness all warrant further investigation as simple to apply, on-farm measures that can be used in conjunction with body condition and subjective assessment to aid management decisions on pig welfare.

The buffalo is an important livestock resource in several countries of South Asia and the Mediterranean regions. However, reproductive efficiency is compromised due to known problems of biological and management origins, such as lack of animal selection and poor nutrition. Under optimal conditions puberty is attained at 15 to 18 months in river buffalo, 21 to 24 months in swamp buffalo and is influenced by genotype, nutrition, management and climate. However, under field conditions these values deteriorate up to a significant extant. To improve reproductive efficiency, several protocols of oestrus and ovulation synchronization have been adopted from their use in commercial cattle production. These protocols yield encouraging pregnancy rates of (30% to 50%), which are comparable to those achieved in buffaloes bred at natural oestrus. The use of sexed semen in buffalo heifers also showed promising pregnancy rates (50%) when compared with conventional non-sexed semen. Assisted reproductive technologies have been transferred and adapted to buffalo but the efficiency of these technologies are low. However, these latest technologies offer the opportunity to accelerate the genetic gain in the buffalo industry after improving the technology and reducing its cost. Most buffaloes are kept under the small holder farming system in developing countries. Hence, future research should focus on simple, adoptable and impact- oriented approaches which identify the factors determining low fertility and oestrus behaviour in this species. Furthermore, role of kisspeptin needs to be explored in buffalo.

Colostrum consists of a number of biologically active proteins and peptides that influence physiological function and development of a neonate. The present study investigated the biological activity of peptides released from first day bovine colostrum through in vitro and in vivo enzymatic digestion. This was assessed for proliferative activity using a human intestinal epithelial cell line, T84. Digestion of the protein fraction of bovine colostrum in vitro was conducted with the enzymes pepsin, chymosin and trypsin. Pepsin and chymosin digests yielded protein fractions with proliferative activity similar to that observed with undigested colostrum and the positive control foetal calf serum (FCS). In contrast trypsin digestion significantly (P<0·05) decreased colostral proliferative activity when co-cultured with cells when compared with undigested colostrum. The proliferative activity of undigested colostrum protein and abomasal whey protein digesta significantly increased (P<0·05) epithelial cell proliferation in comparison to a synthetic peptide mix. Bovine colostrum protein digested in vivo was collected from different regions of the gastrointestinal tract (GIT) in newborn calves fed either once (n=3 calves) or three times at 12-h intervals (n=3 calves). Digesta collected from the distal duodenum, jejunum and colon of calves fed once, significantly (P<0·05) stimulated cell proliferation in comparison with comparable samples collected from calves fed multiple times. These peptide enriched fractions are likely to yield candidate peptides with potential application for gastrointestinal repair in mammalian species.

The objective of the study was to assess the post-weaning growth response of Sahiwal calves reared on four different pre-weaning dietary regimens. The four diets were: (a) whole cow's milk, starter ration (SR; CP = 20%, total digestible nutrients (TDN) = 72%) and Berseem hay (H; Egyptian clover; CP = 21%, TDN = 63%); (b) whole cow's milk + H; (c) milk replacer (MR; reconstituted to supplier specification; Sprayfo®) + SR + H; and (d) MR + H. The protein and fat percentages of reconstituted MR were 2.22 and 1.84, respectively. Milk or MR were fed at the rate of 10% of the calves' body weight (BW) until 56 days of age, and then withdrawn gradually until weaned completely by 84 days of age. The average initial BW of calves in groups A, B, C and D were 56.3 ± 1.0, 47.5 ± 1.0, 40.4 ± 1.0 and 30.3 ± 1.0 kg, respectively. Initially, there were 12 calves in each group with six of each sex; however, one male calf died from each of groups B and C and were not replaced. During the post-weaning period, 13 to 24 weeks, the calves were fed a single total mixed ration ad libitum based on maize, canola meal, wheat straw and molasses containing 16% CP and 70% TDN. Daily feed intake and weekly BW gains were recorded. The data were analyzed by MIXED model analysis procedures using the statistical program SAS. The intake of calves as percent of their BW, feed conversion ratio and cost per kg of BW gain were not different (P > 0.05) across treatments. The daily gain at 24 weeks of age for the pre-weaning treatments A, B, C and D were 746 ± 33, 660 ± 33, 654 ± 33 and 527 ± 33 g/day and the final liveweights of calves were 119 ± 4.2, 102 ± 4.2, 95 ± 4.2 and 75 ± 4.2 kg, respectively. Gains were influenced significantly (P < 0.05) by pre-weaning treatments. The calves fed MR and H only during the pre-weaning period were unable to catch up post weaning with calves on other dietary treatments. The calves fed whole milk from birth at the rate of 10% of liveweight together with concentrates had higher weaning weight and superior growth rate post weaning as well. Thus, pre-weaning feeding was important for higher weaning weights and superior growth rates post weaning.

The objective was to study the growth potential of Sahiwal calves given milk or milk replacer with or without concentrates. For this purpose, forty-eight Sahiwal calves were divided into four groups of 12 animals each with equal sex ratio. In each group, the calves were offered either milk or a milk replacer (MR) at a rate of 10% of their body weight adjusted weekly. In addition to this, calves were fed either a starter ration plus Egyptian clover hay (SR + H) or hay only (H) until the end of trial. The milk or MR was withdrawn gradually from day 56 until animals were weaned completely by day 84. Calves offered milk grew faster than those offered MR (357 ± 9 vs. 162 ± 9 g/day; p < 0.05) and displayed higher weaning weights (51.6 ± 0.8 vs. 35.2 ± 0.8 kg; p < 0.05). The calves offered SR + H grew faster (311 ± 9 vs. 208 ± 9 g/day; p < 0.05) and displayed higher weaning weights (48.7 ± 0.8 vs. 38.1 ± 0.8 kg; p < 0.05) than those fed H alone. Calves offered milk plus SR + H showed the highest growth rate and weaning weights (401 ± 13 g/day and 56.3 ± 1 kg, respectively). The lowest growth rate and weaning weights were observed in calves offered MR and H only (115 ± 13 g/day and 30.3 ± 1 kg, respectively). Calves offered the MR had higher number of scour days than those offered milk (13.5 vs. 3.3). The feeding of whole milk in combination with the starter ration and hay resulted in superior growth rates, higher weaning weights, and healthier calves than the other feeding regimens.

Determining circulating equine insulin concentrations is becoming increasingly important in equine clinical practice and research. Most available assays are optimized for human medicine, but there is strong equine cross-reactivity because of the highly conserved nature of insulin. To identify an accurate and reliable assay for equine insulin, 6 commercial immunoassays were evaluated for precision, accuracy, and specificity. Only 1 assay initially reached the requisite standard: Mercodia Equine Insulin Enzyme-linked Immunosorbent assay (ELISA). Plasma matrix interferences were identified when the provided assay buffer was used with the Siemens Count-a-Coat Insulin radioimmunoassay (RIA) but not when charcoal-stripped equine plasma was used as the diluent. This modified RIA and the Mercodia Equine Insulin ELISA were evaluated further by directly examining accuracy by comparing their results for 18 equine plasma samples with values obtained using liquid chromatography and high-resolution/high-accuracy mass spectrometry (LC-MS). Compared with LC-MS measurements, the modified Siemens Insulin RIA rendered a moderate Lin's concordance coefficient (ρ(c)) of 0.41, whereas the Mercodia Equine Insulin ELISA rendered a very poor ρ(c) of 0.06. This suggests that the Siemens Insulin RIA is appropriate to use for routine evaluations when LC-MS is not available., (Copyright © 2011 Elsevier Inc. All rights reserved.)

It is important to assess ovulation detection performance in commercial dairy herds both to investigate low reproductive performance and to enable herd managers to monitor the effectiveness of their system for detecting ovulations. A method was developed to assess ovulation detection performance that uses limited numbers of strategically collected milk samples, assesses performance over the period when herd managers are making maximal effort to detect ovulations, and when assessing proportions of ovulations detected, accounts for false positive diagnoses of estrus and for cows that have not recommenced postpartum ovulatory cycles. Milk was sampled from cows not diagnosed in estrus early in the breeding program (about d 26 in year-round calving herds and d 22 in seasonal calving herds); milk samples were also collected from cows on the day of insemination. Cows with high milk progesterone concentrations were assumed to have had undetected ovulations and false positive diagnoses of estrus, respectively. The method was successfully implemented in 161 of 167 commercial dairy herds. Positive predictive values (PPV; the proportions of ovulation diagnoses where ovulation was, in fact, imminent) were generally high in both year-round and seasonal calving herds (median values were 0.96 and 0.97, respectively), but 25% of herds had PPV <0.95. Ovulation detection sensitivities (ODS) were low in most year-round calving herds, but many seasonal calving herds had high ODS values; median ODS were 0.73 and 0.94, respectively. However, in 25% of seasonal calving herds, ODS was <0.91. These findings indicate that this method for assessing ovulation detection performance can be successfully implemented in commercial dairy herds with appropriate professional support. The wide range of ODS and the absence of correlation between ODS and PPV suggest that it is possible for managers of many commercial herds in Australia to achieve increased reproductive efficiency through increases in ODS and PPV., (Copyright (c) 2010 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.)

An in vitro bovine mammosphere model was characterized for use in lactational biology studies using a functional genomics approach. Primary bovine mammary epithelial cells cultured on a basement membrane, Matrigel, formed three-dimensional alveoli-like structures or mammospheres. Gene expression profiling during mammosphere formation by high-density microarray analysis indicated that mammospheres underwent similar molecular and cellular processes to developing alveoli in the mammary gland. Gene expression profiles indicated that genes involved in milk protein and fat biosynthesis were expressed, however, lactose biosynthesis may have been compromised. Investigation of factors influencing mammosphere formation revealed that extracellular matrix (ECM) was responsible for the initiation of this process and that prolactin (Prl) was necessary for high levels of milk protein expression. CSN3 (encoding kappa-casein) was the most highly expressed casein gene, followed by CSN1S1 (encoding alphaS1-casein) and CSN2 (encoding beta-casein). Eighteen Prl-responsive genes were identified, including CSN1S1, SOCS2 and CSN2, however, expression of CSN3 was not significantly increased by Prl and CSN1S2 was not expressed at detectable levels in mammospheres. A number of novel Prl responsive genes were identified, including ECM components and genes involved in differentiation and apoptosis. This mammosphere model is a useful model system for functional genomics studies of certain aspects of dairy cattle lactation.

The potential genetic and economic advantage of marker-assisted selection for enhanced production in dairy cattle has provided an impetus to conduct numerous genome scans in order to identify associations between DNA markers and future productive potential. One area of focus has been a quantitative trait locus on bovine chromosome 6 (BTA6) found to be associated with milk yield, milk protein and fat percentage, which has been subsequently fine-mapped to six positional candidate genes. Subsequent investigations have yet to resolve which of the potential positional candidate genes is responsible for the observed associations with productive performance. In this study, we analysed candidate gene expression and the effects of gene knockdown on expression of beta- and kappa-casein mRNA in a small interfering RNA transfected bovine in vitro mammosphere model. From our expression studies in vivo, we observed that four of the six candidates (ABCG2, SPP1, PKD2 and LAP3) exhibited differential expression in bovine mammary tissue over the lactation cycle, but in vitro functional studies indicate that inhibition of only one gene, SPP1, had a significant impact on milk protein gene expression. These data suggest that the gene product of SPP1 (also known as osteopontin) has a significant role in the modulation of milk protein gene expression. While these findings do not exclude other positional candidates from influencing lactation, they support the hypothesis that the gene product of SPP1 is a significant lactational regulatory molecule.

Folding variants of alpha-lactalbumin (alpha-la) are known to induce cell death in a number of cell types, including mammary epithelial cells (MEC). The native conformation of alpha-la however has not been observed to exhibit this biological activity. Here we report that native bovine alpha-la reduced the viability of primary bovine mammary epithelial cells (BMEC) and induced caspase activity in mammospheres, which are alveolar-like structures formed by culturing primary BMEC on extracellular matrix in the presence of lactogenic hormones. These observations suggest a possible role for bovine alpha-la in involution and/or maintaining the luminal space in mammary alveoli during lactation. In addition, co-incubation of bovine alpha-la in an in-vitro mammosphere model resulted in decreased beta-casein mRNA expression and increased alphas1- and kappa-casein mRNA expression. This differential effect on casein expression levels is unusual and raises the possibility of manipulating expression levels of individual caseins to alter dairy processing properties. Manipulation of alpha-la levels could be further investigated for its potential to enhance milk protein expression and/or improve lactational persistency by influencing the balance between proliferation and apoptosis of BMEC, which has a major influence on the milk-producing capacity of the mammary gland.

Secretory characteristics of the ghrelin profile for the pig are still unknown. Our objective was to clarify the mechanisms that influence ghrelin secretion during differing feeding patterns. Pigs were initially fed a commercial pelleted diet offered ad libitum and blood samples collected for 24 h at intervals of 1 h. The pigs were then entrained for 17 days to a twice daily interval feeding regimen (0900-1000 and 1600-1700 h) and blood samples were collected for 12 h (0800-2000 h). This was followed by a similar interval feeding and blood sampling regimen with the 0900-1000 h feeding period being replaced by a sham feed where pigs were shown their usual feed but none offered. During the ad libitum feeding regimen, there was no preprandial rise or postprandial fall in circulating plasma total ghrelin concentration, which remained constant throughout the sampling period. In addition, no preprandial rise or postprandial fall in ghrelin concentrations was observed when pigs were fed either twice or once daily; however, plasma ghrelin concentration rose gradually over the 12-h sampling period during the twice daily feeding regimen and increased further when pigs were fed once per day. This increase in ghrelin levels coincided with an increase in plasma GH and non-esterified fatty acid concentrations and was not associated with either plasma glucose or insulin concentrations. These results suggest that circulating total plasma ghrelin concentrations in the pig appear to be influenced by chronic changes in energy balance rather than the feeding pattern per se.

The concentration of lactoferrin (LTF) in milk varies during lactation, rising sharply during involution. We proposed that LTF might have a regulatory role in involution and investigated its effects in vitro on the viability of bovine mammary epithelial cells (BMEC) and on casein expression in bovine mammospheres. Mammospheres capable of milk protein expression were formed by culturing primary BMEC on extracellular matrix in the presence of lactogenic hormones. Exogenously added LTF decreased beta-casein and kappa-casein mRNA expression in mammospheres while transfection of a short interfering RNA (siRNA) to suppress LTF expression resulted in increased casein mRNA expression. We believe that LTF exerts its effect on casein gene expression by up-regulating interleukin-1beta (IL-1beta) as IL-Ibeta gene expression was elevated in mammospheres treated with LTF. LTF also decreased viability of BMEC grown as monolayers and as mammospheres. Interestingly, LTF was only effective in reducing casein mRNA expression and viability in mammospheres when added at concentrations found during early involution but was inactive when used at concentrations found in milk. We suggest that LTF has a regulatory role during early involution, decreasing casein expression and reducing BMEC viability.

In addition to its role in reproduction, oxytocin has central actions modulating behavioural and hypothalamic-pituitary-adrenal (HPA) axis responses during late pregnancy and lactation. The hypothesis that ovarian hormones modulate the effects of oxytocin on HPA axis activity was studied in 7-day ovariectomised rats receiving oestradiol with or without progesterone replacement and intracerebroventricular (i.c.v) minipump infusion of oxytocin (100 ng/h). In an initial experiment, i.c.v. oxytocin had no effect on basal or restraint-stimulated plasma adrenocorticotrophic hormone (ACTH) and corticosterone concentrations or hypothalamic corticotrophin-releasing factor (CRF) mRNA expression with low oestradiol replacement alone but it had a stimulatory effect in the presence of low oestradiol and progesterone. To investigate further whether oestradiol modulates central actions of oxytocin, rats received low dioestrous (low), pro-oestrous (medium) or pregnancy (high) oestradiol replacement levels, yielding plasma concentrations of < 5, 17.3 +/- 4.5 and 258 +/- 32 pg/ml, respectively, with or without i.c.v. oxytocin. Oestradiol caused dose-dependent increases in basal plasma ACTH and corticosterone concentrations but decreased the ACTH response to restraint stress. In parallel to the changes in basal plasma ACTH, high oestrogen increased basal CRF hnRNA, CRF mRNA in the paraventricular nucleus and pro-opiomelanocortin (POMC) mRNA in the pituitary gland, while decreasing restraint stress-stimulated levels. Intracerebroventricular administration of oxytocin reduced basal and stress-stimulated plasma ACTH, hypothalamic CRF hnRNA (30 min), CRF mRNA and pituitary POMC mRNA (4 h) levels parallel to the increases induced by elevating plasma oestradiol. The present study demonstrates the converse effects of oestradiol on basal and restraint stress-stimulated basal HPA axis activity, and that the ability of central oxytocin to inhibit HPA axis activity depends on the levels of circulating oestradiol.

We evaluated 2 strains of mice for their utility in the investigation of nutritional and molecular regulatory mechanisms of lactation. The lactational performance and milk composition were characterized for an inbred mouse strain, inbred Quackenbush Swiss line 5 (QSi5) selected persistently for fecundity, and a nonselected strain, CBA. The milk yield assessed by changes in BW in response to suckling of sustainable litter sizes for each strain was 3-fold greater (P < 0.001) in QSi5 mice than the CBA strain. The QSi5 mice also produced milk more efficiently (P < 0.001) than CBA mice, despite having the same quantity of mammary tissue per unit of BW. Milk composition did not vary between strains or by stage of lactation, with the exception of lactose concentration, which was greater (P = 0.003) in QSi5 mice. Expression of epsilon-casein was > or = 10-fold greater, and alpha(S1)-casein was > or = 3-fold greater, during mid and late lactation compared with early lactation in both strains, whereas kappa-casein underwent an apparent alteration in posttranslational modifications in both strains from early to mid lactation. Changes in casein composition coincided with an increased susceptibility to proteolytic degradation; hence milk from early lactation may be more readily degraded to facilitate digestion in the neonate. The greater milk synthetic capacity of QSi5 mice over the lactation cycle provides a useful model for studies of nutritional and molecular regulation of lactation.

Because the poor growth performance of intensively housed pigs is associated with increased circulating glucocorticoid concentrations, we investigated the effects of glucocorticoid suppression by inducing a humoral immune response to ACTH on physiological and production variables in growing pigs. Grower pigs (28.6 +/- 0.9 kg) were immunized with amino acids 1 through 24 of ACTH conjugated to ovalbumin and suspended in diethylaminoethyl (DEAE) dextran-adjuvant or adjuvant alone (control) on d 1, 28, and 56. The ACTH-specific antibody titers generated suppressed increases in cortisol concentrations on d 63 in response to an acute stressor (P = 0.002; control = 71 +/- 8.2 ng/mL; ACTH-immune = 43 +/- 4.9 ng/mL) without altering basal concentrations. Plasma beta-endorphin concentrations were also increased (P < 0.001) on d 63 (control = 18 +/- 2.1 ng/mL; ACTH-immune = 63 +/- 7.3 ng/mL), presumably because of a release from negative feedback on the expression of proopiomelanocortin in pituitary corticotropes. Immunization against ACTH did not alter ADG (P = 0.120; control = 1,077 +/- 25; ACTH-immune = 1,143 +/- 25 g) or ADFI (P = 0.64; control = 2,719 +/- 42; ACTH-immune = 2,749 +/- 42 g) and did not modify behavior (P = 0.681) assessed by measuring vocalization in response to acute restraint. In summary, suppression of stress-induced cortisol responses through ACTH immunization increased beta-endorphin concentrations, but it did not modify ADG, ADFI, or restraint vocalization score in growing pigs.

A method for the collection of mammary biopsies developed previously was refined and used to study the endocrine regulation of bovine milk protein gene expression. Our surgical biopsy method used real-time ultrasound imaging and epidural analgesia to enable recovery of a sufficient quantity of mammary tissue from late-pregnant dairy cows for explant culture in vitro. The time of biopsy was critical for prolactin-dependent induction of milk protein gene expression in mammary explants, as only mammary tissue from cows nearing 30 d prepartum was hormone-responsive. This suggests that during the later stages of pregnancy a change in the responsiveness of milk protein gene expression to endocrine stimuli occurred in preparation for lactation. This may relate to the diminution of a putative population of undifferentiated cells that were still responsive to prolactin. Alternatively, the metabolic activity of the tissue had increased to the level whereby the response of the tissue was no longer assessable using this model in vitro.

Hypothalamic corticotropin releasing hormone (CRH) regulates pituitary ACTH secretion and mediates behavioral and autonomic responses to stress, through interaction with type 1 plasma membrane receptors (CRHR1) located in pituitary corticotrophs and the brain. Although the CHRI are essential for ACTH responses to stress, their number in the pituitary gland does not correlate with corticotroph responsiveness, suggesting that activation of a small number of receptors is sufficient for maximum ACTH production. CRH binding and hybridization studies in adrenalectomized, glucocorticoid-treated or stressed rats revealed divergent changes in CRH receptors and CRH1 mRNA in the pituitary, with a reduction in receptor binding but normal or elevated expression of CHR1 mRNA levels. Western blot analysis of CRHR1 protein in pituitary membranes from adrenalectomized rats showed unchanged receptor mRNA levels and increased CRHR1 protein, despite binding down-regulation, suggesting that decreased binding is due to homologous desensitization, rather than reduced receptor synthesis. In contrast, decreased CRH binding following glucocorticoid administration is associated with a reduction in CRHR1 protein, suggesting inhibition of CRH1 mRNA translation. The regulation of CRHR1 translation may involve binding of cytosolic proteins, and a minicistron in the 5'-UTR of the CRHR1 mRNA. It is likely that post-transcriptional regulatory mechanisms that permit rapid changes in CRH receptor activity are important for adaptation of corticotroph responsiveness to continuous changes in physiological demands.

The present study was conducted to determine the pattern of incorporation of dietary EPA and docosahexaenoic acid (DHA) into milk, and to evaluate consequent changes in milk fat composition and sensory characteristics. Fourteen multiparous cows in early lactation were divided into two groups and were offered supplements for 10 d. While individual stalls after each morning milking, one group was offered a mixture of rumen-protected tuna oil (RPTO)-soyabean supplement (2 kg; 30:70, w/w; +RPTO) and the second group was offered the basal ration without RPTO (-RPTO). Both groups grazed together on a spring pasture after supplementation. Feeding supplemental RPTO increased the concentrations of EPA and DHA in milk fat from undetectable levels in -RPTO cows to 6.9 and 10.1 g/kg milk fat respectively. Total n-3 PUFA concentration in milk fat was increased three- to fourfold by tuna-oil supplementation (8.4 to 32.0 g/kg milk fat). There were no significant effects on milk production (35.4 v. 33.9 l/d), milk protein (28.2 v. 30.1 g/kg) or milk fat (36.2 v. 40.4 g/kg for -RPTO and +RPTO respectively). The concentration of total saturated fatty acids in milk fat was significantly reduced (568 v. 520 g/kg total fatty acids) and there was a 17 % reduction in the atherosclerotic index of milk after tuna-oil supplementation. Untrained consumer panellists (n 61) rated milk from both groups of cows similarly for taste and smell. We conclude that it is possible to enrich milk with n-3 PUFA without deleterious effects on yield, milk composition or sensory characteristics.

Saliva was collected from neonatal piglets in order to assess the secretory dynamics of cortisol in the newborn piglet. Saliva was collected from two litters of piglets (n = 16) from multiparous sows at 07.00, 11.00, 15.00 and 19.00 h on days 0, 1, 2, 4, 6, 10, 14, 18, 22, 26 and 30 post partum. Salivary cortisol concentration was determined by radioimmunoassay. Salivary cortisol concentration was high perinatally and declined with age. The frequency of escape attempts and vocalisation intensity during collection, as well as the similarity in salivary cortisol concentration on day 22 in control piglets and piglets that had provided samples repeatedly from day 0 suggested that this was not an adaptive response. A circadian pattern was evident in female pigs from day 6, whereas males displayed a similar pattern from day 10. The collection of saliva in the piglet provides a viable alternative to blood sampling for the assessment of a glucocorticoid circadian rhythm., (Copyright 2002 S. Karger AG, Basel)

Eighty two multiparous Holstein cows were blocked by genetic merit (high vs. low) and assigned to one of two treatments [high rumen-undegradable protein (RUP): rumen-degradable protein (RDP) vs. low RUP: RDP] from d 21 before to d 150 after calving to study the effects of these treatments on production and reproductive performance. Diets were isonitrogenous (dry cow 10.5% crude protein; lactating cow 19.3%), isoenergetic (dry cow 10.0 MJ of metabolizable energy (ME); lactating cow 10.9 MJ of ME) and fed as total mixed rations. Feeding more RUP significantly increased dry matter intake and milk yield, reduced body tissue mobilization, and lowered concentrations of serum nonesterified fatty acids (NEFA) and plasma urea. Expression of estrus at first ovulation was improved, first service conception rate was higher, and calving to conception interval was shorter for the high RUP group. Cows of high genetic merit produced more milk, mobilized more body tissue, and had higher concentrations of plasma growth hormone. The dry matter intake and concentrations of blood metabolites did not significantly differ with genetic merit. Expression of estrus at first ovulation was significantly lower for cows of high genetic merit. Serum NEFA concentrations were significantly higher, and estrus was not observed at first ovulation for cows of higher genetic merit fed the low RUP diet. The interaction between dietary RUP and genetic merit was not significant for other measures of performance or fertility. Feeding a low RUP: high RDP diet had negative effects on some aspects of production and reproductive performance. The effects of diet on NEFA concentrations and estrus display were greater in cows of high genetic merit, indicating that potential interactions should be evaluated in future reproductive studies involving protein and fertility.