The effects of maternal calcium status during late gestation on lamb growth and survival in twin-bearing Merino ewes grazing pasture.

Our project aimed to investigate whether low maternal calcium status during late gestation decreased weight, growth and survival of lambs born to twin-bearing Merino ewes grazing pasture. On day 130 of gestation, approximately a 10 mL blood sample was collected from 108 twin-bearing Merino ewes. Blood pH and calcium ion concentration were measured in whole blood and serum was analysed for calcium, magnesium, phosphate, beta hydroxy butyrate, non-esterified fatty acids and cholesterol. Six to 18 h after birth and at 49 days of age, lamb live weight was recorded. Ewe serum calcium level at day 130 of gestation were divided into tertiles (1.58-2.13, 2.14-2.30, and 2.31-2.99 mmol/L) with ewes classified as either Low, Med or High (n = 36 per group). A general linear mixed model was used to analyse the data. Survival to 1, 2, 3 and 7 days of age was lower for lambs born to ewes in the Low calcium group compared with those born in the Med and High calcium groups (P < 0.05). At day 49 of age, lamb survival tended to be lower for lambs in the Low (73.6%) compared with Med (86.8%) and High (85.3%) calcium groups (P < 0.1). There were no differences in lamb body weight (P > 0.05). Calcium levels of ewes in the Low calcium group were, on average, indicative of subclinical hypocalcaemia, with this group of ewes also having the lowest lamb survival rate, suggesting that maternal subclinical hypocalcaemia reduces lamb survival. Further studies, particularly on improving maternal calcium levels, are warranted.

Supplementing pregnant Merino ewes with caffeine to improve neonatal lamb thermoregulation and viability.

Percentages of lamb mortalities prior to weaning are a significant and persistent problem for the Australian sheep industry. Maternal caffeine supplementation reduced stillbirths and improved viability in piglets; however, the efficacy of caffeine in enhancing viability and lamb survival is equivocal. The aim of this study was to determine the optimal concentration and duration of maternal caffeine supplementation to improve lamb viability; time to stand and suck with consumption of colostrum occurring, as well as survival to weaning. Multiparous Merino ewes were supplemented with either 0 (CTL), 10 (C10) or 20 mg/kg (C20) body weight (BW) caffeine in feed after day 120 of gestation (dG), or 20 mg/kg BW (LC20) caffeine from dG 142 until parturition. Ewes were housed indoors in individual lambing pens from dG 130 to 72 -h post-partum (pp). Values for pp ewe and lamb variables were analysed using a generalised linear mixed model in IBM SPSS version 25. While ewes within the CTL, C10 and LC20 groups consumed more caffeine compared to C20 ewes (P =  0.001), lambs of C20 ewes had greater rectal temperatures at 20 -h pp (P =  0.021), greater 4 -h serum IgG concentrations (P =  0.041), a longer latency to first sucking bout (P =  0.030), and a greater number of sucking attempts (P =  0.044) compared to lambs from CTL, C10 and LC20 ewes. These results indicate that caffeine supplementation during late-gestation stimulates neonatal lambs as a result of increased sucking (4 -h serum IgG) and increased temperatures within the first 24 -h.

Effect of feed grade L-methionine on growth performance and gut health in nursery pigs compared with conventional DL-methionine.

Two experiments were conducted to test if supplementation of LMET has beneficial effects on growth performance and gut health in nursery pigs compared with DL-Met. In Exp. 1, 168 pigs in 56 pens were randomly allotted to 7 dietary treatments for 20 d, including a basal diet (BD; 55% of the NRC requirement for Met), the BD+0.048% L-Met or DL-Met (70% of the NRC requirement), the BD+0.096% L-Met or DL-Met (85% of the NRC requirement), and the BD+0.144% L-Met or DL-Met (100% of the NRC requirement). Body weight and feed disappearance were recorded every 5 d for computation of growth performance. In Exp. 2, 20 individually housed nursery pigs were randomly allotted to 2 dietary treatments for 20 d: DML (0.16% Met from the BD+0.145% supplemental DL-Met) or LMET (0.16% Met from the BD+0.145% supplemental L-Met). Both diets had Met meeting 95% of the NRC requirement. Duodenum samples from all pigs were collected at the end of the trial to evaluate morphology and redox status. In Exp. 1, during the entire 20 d, pigs fed diets supplemented with L-Met tended to have greater (P=0.087) ADG and reduced (P<0.01) plasma urea nitrogen (PUN) than pigs fed diets supplemented with DL-Met. The relative bioavailability (RBA) of L-Met to DL-Met for ADG and G:F was 143.8 and 122.7%, respectively. In Exp. 2, pigs fed a diet supplemented with L-Met had duodenum tissue with greater (P<0.05) concentrations of glutathione (GSH) and greater villus height and width as well as lower (P<0.05) concentrations of protein carbonyl compared with pigs fed DL-Met. Overall, compared with DL-Met, the use of L-Met as a source of supplemental Met in nursery pig diets enhanced duodenum villus development in association with reduced oxidative stress and improved GSH. The beneficial effects of supplementing L-Met compared to DL-Met in gut of nursery pigs resulted in a potential enhancement of ADG and reduction of PUN.

Efficacy of dietary spray dried plasma protein to mitigate the negative effects on performance of pigs fed diets with corn naturally contaminated with multiple mycotoxins.

The ability of spray dried plasma protein (SDPP) to reduce the negative effects of multiple mycotoxins from naturally contaminated corn on weaned pig performance and health was investigated (n = 180; 6.84 ± 0.11 kg). For 12 d after weaning, pigs were fed phase 1 nursery diets with either 0% SDPP (PP0) or 6% SDPP (PP6). After 12 d, pigs were fed phase 2 diets for 3 wk. Pigs fed PP0 in phase 1 continued to be fed a phase 2 diet with no SDPP (PP0/PP0) or were fed a diet including corn naturally contaminated with multiple mycotoxins (M), labeled PP0/PP0M. Pigs fed SDPP in phase 1 were fed either a diet with no SDPP (PP6/PP0), a diet with M and no SDPP (PP6/PP0M), a diet with M and 3% SDPP (PP6/PP3M), or a diet with M and 6% SDPP (PP6/PP6M). During phase 1, pigs fed PP6 had increased (P < 0.05) ADG, ADFI, and G:F, whereas immunological parameters were not altered. During phase 2, pigs consuming PP0/PP0M had reduced ADG (P < 0.01) and ADFI (P < 0.05) in contrast to pigs fed PP0/PP0, whereas the performance of pigs fed PP6/PP0M was intermediate to pigs fed PP0/PP0M and PP6/PP0. The ADG and ADFI did not differ for pigs fed PP0/PP0M and PP6/PP0M during phase 2. Performance of pigs fed PP6/PP3M in contrast to pigs fed PP6/PP0M during phase 2 did not differ; however, these pigs had lower (P < 0.05) tumor necrosis factor α and tended (P = 0.094) to have lower DNA damage. During phase 2, ADG and ADFI of pigs fed PP6/PP6M did not differ from pigs fed PP6/PP0M, but G:F tended (P = 0.067) to be increased in pigs fed PP6/PP6M. Over the entire study period, pigs fed PP0/PP0M had reduced (P < 0.05) ADG and tended (P = 0.067) to have reduced ADFI. During this time, pigs fed PP6/PP0M tended to have greater ADG and ADFI (P = 0.093 and P = 0.067, respectively) compared with pigs fed PP0/PP0M. Overall, feeding a diet with SDPP improved growth performance and feed intake of young pigs directly after weaning. Feeding multiple M had a negative impact on growth performance of pigs during this trial. This response was more significant when pigs were not fed SDPP in phase 1. Overall, when combining phase 1 and 2 performance data, daily gain and feed intake tended to be reduced when pigs were not fed 6% SDPP in phase 1. This study indicates that the composition of diets fed immediately after weaning may be important for pigs that subsequently are under a M challenge.

Supplemental nucleotides high in inosine 5'-monophosphate to improve the growth and health of nursery pigs.

This study was conducted to determine the ability of a supplemental nucleotide mixture high in inosine 5'-monophosphate (5'IMP) to enhance pig growth and health after weaning. Pigs (n = 120 and 7.3 ± 0.1 kg) were allotted at weaning to phase 1 diets (3.3 Mcal ME/kg, 22.4% CP, and 1.34% standardize ileal digestible [SID] Lys) supplemented with 0.0, 0.2, 0.5, or 1.0 g/kg of a nucleotide additive. After 7 d, pigs were fed phase 2 diets (3.3 Mcal ME/kg, 21.3% CP, and 1.20% SID Lys) for 21 d with the same additive levels. Growth performance was measured, blood samples were collected for analysis of immune responses and oxidative stress, and fecal scoring was completed. During phase 1, ADG, ADFI, and G:F linearly increased (P < 0.05) as dietary nucleotides increased. During phase 2, ADFI increased linearly (P < 0.05). Over the entire 28-d trial, ADG and ADFI increased linearly (P < 0.05) as nucleotide content increased. Immune responses were not altered during phase 1. At the end of phase 2, IgA showed a quadratic effect (P < 0.05) with the lowest concentrations at 0.2 and 0.5 g/kg of the nucleotide additive whereas IgM changed cubically (P < 0.05) with the lowest concentration at 0.5 g/kg. The cytokine tumor necrosis factor-α tended to decrease linearly (P = 0.093) as nucleotide content increased whereas the marker for oxidative DNA damage, 8-hydroxy-deoxyguanosine, tended to have a quadratic effect (P = 0.064) with the lowest levels of damage in pigs fed 0.5 g/kg. On d 8 after changing from the phase 1 to phase 2 diets, fecal diarrhea scores tended to be lowest (P = 0.072) when pigs were fed 0.5 g/kg of the nucleotide additive. Overall, 1.0 g/kg of the nucleotide additive provided the most benefit to the growth performance of nursery pigs. However, 0.5 g/kg of the nucleotide additive reduced immune responses and oxidative stress. In conclusion, the addition of a nucleotide mixture high in 5'IMP to the diet of young pigs may be beneficial to enhance growth performance and reduce postweaning stress.

Oocyte maturation and embryo survival in nulliparous female pigs (gilts) is improved by feeding a lupin-based high-fibre diet.

Inclusion of high levels of the high-fibre ingredient sugar-beet pulp in pre-mating diets has been shown to increase gonadotrophin concentrations and improve oocyte quality in nulliparous pigs (gilts). This study evaluated the effects of two alternative fibre sources on reproductive performance in gilts. Gilts received one of three diets from 3 weeks before puberty stimulation until Day 19 of the first oestrous cycle: control (39 g kg⁻¹ fibre), bran (500 g kg⁻¹ wheat bran, 65 g kg⁻¹ fibre) or lupin (350 g kg⁻¹ lupin, 118 g kg⁻¹ crude fibre). Diet did not affect circulating LH concentrations or ovarian follicle size. However, a higher percentage of oocytes collected from lupin-supplemented gilts reached metaphase II in vitro compared with those collected from bran-fed or control gilts (89±5% versus 72±5% and 66±5%, respectively; P<0.05). Furthermore, in a second experiment, gilts fed the same lupin-based diet before mating had improved embryo survival (92±5%) on Day 28 after mating compared with control gilts (76±4%; P<0.05). Therefore, feeding a high-fibre diet before mating can improve oocyte quality in gilts without changes in circulating LH, but this effect is dependent on the fibre source.

Bullous pemphigoid antigen localization suggests an intracellular association with hemidesmosomes.

Autoantibodies to a normal component of stratified squamous epithelia, the bullous pemphigoid antigen (BPA), are synthesized in patients with the disease bullous pemphigoid. We have used these sera to study the distribution of BPA in vivo and in vitro. At low magnification, indirect immunofluorescent staining for BPA is linear at the basement membrane zone (BMZ) of skin and many other epithelial tissues. At higher magnification however, we observed a punctate staining pattern for BPA which was regular in appearance and suggested localization of BPA to discrete structures at the BMZ. Subsequent immunoelectron microscopy using both peroxidase and colloidal gold labeling techniques with patients' sera or IgG, revealed that BPA is associated with hemidesmosomes--putative adhesion structures at the BMZ, based on their similarity in ultrastructure to desmosomes. More specifically BPA was immunolocalized to the cytoplasmic face of hemidesmosomes and was not observed extracellularly in the basement membrane. In stratifying and nonstratifying cultures of rat keratinocytes, BPA is expressed intracellularly and not in the cell-derived matrix, unlike other known basement membrane components. These cells also synthesize BPA in vitro, and immunoprecipitation from metabolically labeled cultures revealed a 220 kD polypeptide under reducing conditions. From these observations we conclude (1) that BPA is a 220 kD polypeptide component either of or associated with hemidesmosomes, and (2) that it is localized intracellularly both in vivo and in vitro. We suggest that BPA is not normally a lamina lucida component, but that it may form part of a linkage between the cytoskeleton and the basement membrane.

Fibronectin distribution during the development of fetal rat skin.

Fibronectin distribution during fetal rat skin development has been studied immunocytochemically at the light and electron microscope level from 16 days of gestation to birth. The dermal-epidermal junction, the dermis, and connective tissue around developing muscle were shown by light microscopy to be heavily stained throughout this period. The development of hair follicles from about 18 days onward was not associated with any consistent change in fibronectin distribution. The heavy staining of the upper dermis was associated with a high density of mesenchymal cells, and immunoelectron microscopy revealed fibronectin on the surface of many of these cells and in association with the surrounding fine collagen fibrils. At the dermal-epidermal junction, both follicular and interfollicular, fibronectin was localized mainly in the plasma membrane and lamina lucida regions of the basement membrane, and there was also staining associated with the underlying fine collagen fibrils. These observations are further evidence for the proposed role of fibronectin as a mediator of the cell-matrix interactions which are of importance for tissue development and maintenance.

Decision aid development for use in ambulatory health care settings.

An approach is presented using interactive microcomputers for the development of diagnostic decision aids applicable to some complaints encountered in ambulatory care. The central feature of the descriptive phase of the approach is the use of the underlying (and perhaps dynamic) state of patient health. The central feature of the prescriptive phase of the approach is quick, simple assessment which produces a set of nondominated diagnostic tests, the selection of which is biased by the subjectively determined disease(s) that the diagnostician wishes to rule out or confirm. We present an application of the approach to the complaint, "diarrhea of recent onset in adults," discuss the hardware/software implementation, and summarize preliminary evaluation results.

Fibronectin distribution in epithelial and associated tissues of the rat.

Specific antiserum was used to investigate the distribution of the extracellular glycoprotein, fibronectin, in rat skin and tongue tissue by light and electron microscopy with immunofluorescence and immunoperoxidase techniques. We conclude that fibronectin is absent from stable, differentiated parts of tissues, such as the sebaceous glands or the matrix, medulla, cortex, and cuticles of the hair and the inner and outer root sheaths, or even in tissues in which there is some cell movement, such as the epidermis. It is, however, characteristic of sites at which cell division is occurring in contact with an extracellular scaffolding, such as basement membrane or loose connective tissue. Conspicuous examples were in the glassy membrane and connective tissue sheath associated with the follicular epithelium, the basement membrane underlying vascular endothelial cells, the connective tissues surrounding and investing nerve and muscle fibre bundles, and the dermal connective tissue where fibronectin was often associated closely with collagen fibres. At the basement membrane of the dermal/epidermal junction, fibronectin occurred at the plasma membrane of the basal cells and in the lamina lucida area. There was no correlation with specific areas of cell-substrate adhesion, such as the hemidesmosomes. The endoplasmic reticulum of fibroblasts stained strongly suggesting that these cells represent a major site of synthesis.

The interaction of 3,3',4',5-tetrachlorosalicylanilide with phosphatidylcholine bilayers.

1. The interaction of the germicide 3,3',4',5-tetrachlorosalicylanilide (T4CS) with vesicles and dispersions of egg phosphatidylcholine has been studied by gel permeation chromatography, electron microscopy, electron spin resonance spin labelling and ion permeability measurements. 2. Incorporation of T4CS into vesicles of egg phosphatidylcholine gives rise to a large increase in the permeability rate of the paramagnetic cation N,N-dimethyl-N-(1'-oxyl-2',2',6',6'-tetramethyl-4'-piperidyl)-2-hydroxyethylammonium chloride through the lipid bilayer but has no significant effect on the vesicle sizes as measured by gel permeation chromatography or electron microscopy. 3. ESR studies using a spin-labelled fatty acid have demonstrated the presence of two different environments for the spin label when T4CS is incorporated into phosphatidylcholine bilayers. These two environments are identified as (a) highly ordered areas of the bilayer, rich in T4CS and (b) areas with very similar ordering to that in pure egg phosphatidylcholine. 4. The effectiveness of very low concentrations of the germicide in increasing vesicle permeability is explained in terms of its clustering to give rigid patches, rich in T4CS, rather than being evenly distributed throughout the bilayer. It is proposed that the increased ion permeability arises from leakage at the interfaces between the rigid and flexible regions of the lipid bilayer. 5. Comparisons between the effective levels of T4CS in phosphatidylcholine vesicles and its minimum inhibitory concentration with a Gram-positive bacterium confirm the validity of phospholipid vesicles as a model for studies of germicidal activity.