Short communication: Red deer (Cervus elaphus) colostrum during its transition to milk.

We studied changes in chemical composition, somatic cell count, and immunoglobulin G (IgG) and M (IgM) content in red deer (Cervus elaphus) colostrum during the transition to milk at different times after parturition (<5 h, 24 h, 48 h, 2 wk, and 4 wk). The production level was higher at 2 and 4 wk of lactation than during the first day after parturition, with intermediate values at 48 h postpartum. Fat content did not vary during the study period. However, total protein and casein contents were particularly high in the initial 5 h after parturition, decreasing to approximately 50% after 24 h postpartum. Conversely, lactose concentration was low in the beginning (<5 h), increasing gradually throughout the study. Similarly, dry matter dropped during the first 24 h and then remained constant throughout the study. Urea content decreased during the study, showing a slight recovery at 4 wk. Somatic cell count was higher during the first hours after parturition and gradually decreased throughout the study period. The IgG content was higher before 5 h postpartum than at 24 h postpartum. After 5 h, the level of IgG decreased progressively until it reached 0.18 mg/mL at 4 wk of lactation. We observed a similar pattern for IgM content, but it decreased more quickly than IgG and was not detected after 2 wk. In the case of deer, milk should be considered transitional from 24 to 48 h after parturition, and samples collected after 2 wk can be considered mature milk.

Improving appearance and microbiologic quality of broiler carcasses with an allostatic modulator.

An important priority of poultry producers is to guarantee animal welfare during animal production; however, broilers are exposed to unavoidable chronic stress (also known as allostasis) when they are captured, caged, and transported to the processing plant. This antemortem management causes allostatic load, animal injuries, and poor carcass quality. The aim of the present study was to evaluate the effects of an allostatic modulator (AM) on antemortem stress by measuring the appearance and microbiological quality of broiler carcasses. The AM consisted of a liquid formula containing ascorbic acid, acetyl salicylic acid, and electrolytes, administered orally 48 h before shipment to the processing plant. A total of 600 chickens (49-days-old) were used under a factorial arrangement 2 × 2 × 2 [2 commercial hybrid lines, 2 feed withdrawal programs (10 and 16 h), and 2 water treatments (control and AM)]. Each treatment included 25 chickens per pen and was carried out in triplicate. The broilers were shipped, slaughtered, and processed in a commercial processing plant where carcass defects (bruises and broken bones caused by antemortem management), crop pH, and carcass bacterial counts were evaluated in all experimental groups. Broilers under AM treatment showed a reduction in carcass defects (P = 0.015), crop pH (P = 0.0001), coliforms counts (P = 0.014), and total aerobic mesophilic bacteria (P = 0.0001) when compared to the control treatment. The present study indicates that the AM can be used to improve carcass quality in broilers. Our study provides a novel and economic alternative to reduce the allostatic load in broilers.

Influence of feed form and source of soybean meal on growth performance, nutrient retention, and digestive organ size of broilers. 2. Battery study.

Two experiments were conducted to determine the apparent ileal digestibility (AID) of the amino acids (AA) of 4 commercial soybean meals (SBM) from the United States (USA-1, 48.1% CP and USA-2, 46.2% CP), Brazil (BRA, 47.6% CP), and Argentina (ARG, 46.3% CP) and the effects of the inclusion of these SBM in diets in mash, crumble, or pellet form on growth performance, total tract apparent retention of nutrients, and digestive organ size in broilers reared in cages from 1 to 25 d of age. In experiment 1, the AID of Lys was higher (P < 0.05) for the USA-2 than for the BRA SBM, with the SBM from USA-1 and ARG being intermediate. In experiment 2, 12 diets were arranged as a 3 × 4 factorial with 3 feed forms (mash, crumbles, and pellets) and the 4 sources of SBM used in experiment 1. The feeds were isonutritive and the AID of the AA of the SBM obtained in experiment 1 was used for diet formulation. Broilers fed mash had lower (P < 0.001) ADFI and ADG and poorer (P < 0.001) feed-to-gain ratio than broilers fed crumbles or pellets but source of SBM did not affect growth performance. Nitrogen retention was higher (P < 0.01) in birds fed mash than in birds fed crumbles or pellets at all ages. The total tract apparent retention of nutrients was lower (P < 0.05) for the BRA and ARG SBM diets than for the USA-1 and USA-2 SBM diets. Gizzard empty relative weight (% BW) was higher and gizzard pH lower for broilers fed mash than for broilers fed crumbles or pellets (P < 0.001). The results indicate that crumbling or pelleting of the diets improved growth performance of broilers from 1 to 25 d of age. Diets formulated with analyzed rather than calculated AID of AA of the SBM sources resulted in similar broiler performance.

Influence of feed form and source of soybean meal of the diet on growth performance of broilers from 1 to 42 days of age. 1. Floor pen study.

In total, 3,120 broilers were used to study the effects of feed form and source of soybean meal (SBM) of the diet on growth performance. From 1 to 21 d of age, there were 12 treatments arranged factorially with 3 feed forms (mash, crumbles, and pellets) and 4 commercial sources of SBM that differed in the CP content [48.1 and 46.2% CP from the United States (USA-1 and USA-2), 47.6% CP from Brazil (BRA), and 46.3% CP from Argentina (ARG)]. From 21 to 42 d of age, diets were fed as pellets. Diets were formulated assuming that all SBM had similar digestible amino acid content per unit of CP. From 1 to 21 d of age, chicks fed crumbles or pellets had higher (P < 0.001) ADG than chicks fed mash. Also, chicks fed pellets had better (P < 0.001) feed-to-gain ratio (F:G) than chicks fed crumbles, and both were better than chicks fed mash. However, from 21 to 42 d of age, F:G was best (P < 0.001) for chicks previously fed mash. For the entire experimental period, broilers that were fed crumbles or pellets from 1 to 21 d of age had higher (P < 0.001) ADG than broilers that were fed mash. Also, broilers that were fed pellets had better (P < 0.05) F:G than broilers fed mash, with broilers fed crumbles being intermediate. Broilers fed the USA-2 meal had higher (P < 0.01) ADG than broilers fed the BRA or the ARG meals, with broilers fed the USA-1 meal being intermediate. Feed efficiency tended (P = 0.07) to be hindered in broilers fed the BRA meal. The results show that pelleting improved growth performance of broilers from 1 to 42 d of age with effects being less evident at 42 d than at 21 d of age. Source of SBM affected growth performance suggesting the need for a better control of chemical composition and quality of this ingredient before diet formulation.

Effects of maternal age and offspring sex on milk yield, composition and calf growth of red deer (Cervus elaphus).

Differential maternal allocation theory states that mothers will invest more heavily in the offspring sex that will secure higher reproductive output. Senescence theory is concerned with the gradual deterioration of physiological function with age. We analysed the offspring sex-dependent response of calf growth and milk traits to mother age in an Iberian population of captive red deer (Cervus elaphus) using a 22 year time series longitudinal data set. Previous studies revealed that there was little evidence for the differential allocation theory on milk traits and that most studies lacked proper control for confounding factors. Our results indicated that (i) calf growth was offspring male-biased, negatively affected by mother age and positively influenced by mother weight and parity, and (ii) there was no support for differential allocation offspring sex-dependence in milk traits (yield, energy density, fat, protein and lactose content). Our findings suggest that maternal allocation responds to offspring energy requirements, which are mainly driven by offspring body weight, and contingent on mother age and weight and previous maternal reproductive effort.

Water sprinkling as a tool for heat abatement in farmed Iberian red deer: Effects on calf growth and behaviour.

Climatic models predict scenarios in which ambient temperature will continue increasing worldwide. Under these climatic conditions, fitness and animal welfare of many populations are expected to suffer, especially those that live in captive or semi-natural conditions, where opportunities of heat abatement are limited. We undertook an experimental design to assess the effect of heat abatement that water sprinkling might have on Iberian red deer calf growth and behaviour from birth to weaning (135 days). One group of ten mother-calf pairs lived on plots with water sprinkling (treatment) available during summer's hottest time of the day, while the control group (nine mother-calf pairs) occupied plots with no available water sprinkling. Treatment and control groups were fed ad libitum and swapped between plots every seven days to minimise any plot effect. Body weight was monitored weekly and individual behaviour was recorded once or twice a week at mid-day. We observed that calves had showers under the sprinklers and wallowed in mud puddles. The results clearly indicated that calves of the treatment group showed a significant increase in body weight at weaning in comparison with the control group, with no differences between sexes (treatment: male = 56.5 kg, female = 50.3 kg; control: male = 50.3 kg, female = 46.5 kg). Mother weight and mother age effects were negligible on calf body weight at weaning. The heavier the mother the faster was the rate of growth of its offspring, irrespective of calf sex. The model indicated that although males grew significantly slower than female calves in the control group, males grew faster than females when exposed to the treatment. Calves of the treatment group spent less time drinking, less time in the shade, similar time eating and more time in motion than calves of the control group. There were no behavioural differences between calf sexes of treatment and control groups. The results indicate the importance of providing animals with opportunities of heat abatement in hot environments to improve animal growth and welfare in farmed Iberian red deer.

Heat stress reduces growth rate of red deer calf: Climate warming implications.

Climate models agree in predicting scenarios of global warming. In endothermic species heat stress takes place when they are upper their thermal neutral zone. Any physiological or behavioural mechanism to mitigate heat stress is at the cost of diverting energy from other physiological functions, with negative repercussions for individual fitness. Tolerance to heat stress differs between species, age classes and sexes, those with the highest metabolic rates being the most sensitive to stressing thermal environments. This is especially important during the first months of life, when most growth takes place. Red deer (Cervus elaphus) is supposedly well adapted to a wide range of thermal environments, based on its worldwide distribution range, but little is known about the direct effect that heat stress may have on calf growth. We assessed the effect that heat stress, measured by heat stress indices and physical environment variables (air temperature, relative air humidity, wind speed and solar radiation), have on calf and mother body weights from calf´s birth to weaning. We used 9265 longitudinal weekly body weight records of calf and mother across 19 years in captive Iberian red deer. We hypothesised that (i) heat stress in hot environments has a negative effect on calf growth, especially in males, as they are more energetically demanding to produce than females; and that (ii) the body weight of the mother through lactation should be negatively affected by heat stress. Our results supported hypothesis (i) but not so clearly hypothesis (ii). By weaning (day 143) calves growing under low heat stress environment grew up to 1.2 kg heavier than those growing in high heat stress environment, and males were more affected by heat stress than females. The results have implications in animal welfare, geographical clines in body size and adaptation to climate change.

What do rates of deposition of dental cementum tell us? Functional and evolutionary hypotheses in red deer.

Cementum is a bone connective tissue that provides a flexible attachment for the tooth to the alveolar bone in many mammalian species. It does not undergo continuous remodelling, unlike non-dental bone, which combined with its growth pattern of seasonal layering makes this tissue uniquely suitable as a proxy for tracking changes in body repair investment throughout an animal´s life. We tested functional and sexual selection hypotheses on the rate of cementum deposition related to the highly polygynous mating strategy of red deer. We used a sample of 156 first lower molars from wild Scottish red deer of known age between 1 and 17 years old, approximately balanced by sex and age class. Cementum deposition on the inter-radicular pad increased with age at a constant average rate of 0.26 mm per year, with no significant differences between sexes. Cementum deposition was independent of (i) tooth wear, other than that associated with age, and (ii) enamel and dentine micro-hardness. The results partially supported the hypothesis that the main function of cementum is the repositioning of the tooth to maintain opposing teeth in occlusion. However, teeth that had more wear or males´ teeth that had faster rates of tooth wear than those of females did not present the expected higher rates of cementum deposition.

Influence of the main cereal and feed form of the diet on performance and digestive tract traits of brown-egg laying pullets.

The influence of the main cereal and feed form of the diet on performance and digestive tract traits was studied in 576 brown-egg laying pullets from 1 to 120 d of age. From 1 to 45 d of age, 4 diets arranged factorially with 2 cereals (corn vs. wheat) and 2 feed forms (mash vs. pellets) were used. Each treatment was replicated 6 times (24 pullets per replicate). From 46 to 120 d of age, all diets were offered in mash form and the only difference among diets was the cereal used. Cumulatively, pullets fed the corn diets had higher BW gain (P < 0.05) but similar feed conversion ratio as pullets fed the wheat diets. From 1 to 45 d of age, pullets fed pellets consumed more feed (P < 0.001) and had higher BW gain (P < 0.001) than those fed mash. Most of the beneficial effects of pelleting on productive performance were still evident at 120 d of age. At 45 d of age, gizzard weight (g/kg of BW) was higher (P < 0.01) in pullets fed corn than in pullets fed wheat diets. Feeding pellets reduced the relative weight of the digestive tract and the gizzard (P < 0.001) as well as the length (cm/kg of BW) of the small intestine (P < 0.01) at both ages. The pH of gizzard contents at 120 d of age was not affected by cereal but was lower in pullets that were fed mash from 1 to 45 d of age (P < 0.01). We conclude that wheat can be used in substitution of corn in pullet diets with only a slight reduction in BW gain and that feeding pellets from 1 to 45 d of age increased BW gain and pH of the gizzard and reduced the relative weight of the gizzard and the length of the gastrointestinal tract at 120 d of age.

Effect of main cereal of the diet and particle size of the cereal on productive performance and egg quality of brown egg-laying hens in early phase of production.

A total of 960 Lohmann Brown laying hens were used to study the effects of the main cereal of the diet and geometric mean diameter (GMD) of the cereal on productive performance and egg quality from 20 to 48 wk of age. The experiment was a completely randomized design with 6 treatments arranged factorially with 2 cereals (dent corn vs. durum wheat) and 3 GMD of the cereal (hammer-milled to pass through 6-, 8-, or 10-mm screens). Each treatment was replicated 8 times (20 hens per replicate). All diets were formulated to be isonutritive and contained 50% of either corn or wheat. Productive traits were recorded every 4 wk and egg quality was measured at 48 wk of age. The only significant effect detected was for feed intake (P < 0.05) that was greater for hens fed coarse-ground cereals (10-mm screen) than for hens fed medium and fine-ground cereals (8- or 6-mm screen). None of the egg quality parameters studied were influenced by dietary treatment. We conclude that neither type of cereal nor GMD affected productive performance or egg quality of young brown hens, except for feed intake that increased with the coarser particle size.

Meat quality of farmed red deer fed a balanced diet: effects of supplementation with copper bolus on different muscles.

Supplementation with copper (Cu) improves deer antler characteristics, but it could modify meat quality and increase its Cu content to levels potentially harmful for humans. Here, we studied the effects of Cu bolus supplementation by means on quality and composition of sternocephalicus (ST) and rectus abdominis (RA) muscles (n=13 for each one) from yearling male red deer fed with a balanced diet. Each intraruminal bolus, containing 3.4 g of Cu, was administered orally in the treatment group to compare with the control group. Meat traits studied were pH at 24 h postmortem (pH24), colour, chemical composition, cholesterol content, fatty acid (FA) composition, amino acid (AA) profile and mineral content. In addition, the effect of Cu supplementation on mineral composition of liver and serum (at 0 and 90 days of treatment) was analysed. No interactions between Cu supplementation and muscle were observed for any trait. Supplementation with Cu increased the protein content of meat (P<0.01). However, Cu content of meat, liver and serum was not modified by supplementation. In fact, Cu content of meat (1.20 and 1.34 mg/kg for Cu supplemented and control deer, respectively) was much lower in both groups than 5 mg/kg of fresh weight allowed legally for food of animal origin. However, bolus of Cu tended to increase the meat content of zinc and significantly increased (P<0.05) the hepatic contents of sodium and lead. Muscles studied had different composition and characteristics. The RA muscle had significantly higher protein content (P<0.001), monounsaturated FA content (P<0.05) and essential/non-essential AA ratio (P<0.01) but lower pH24 (P<0.01) and polyunsaturated FA content (P=0.001) than the ST muscle. In addition, RA muscle had 14.4% less cholesterol (P=0.001) than ST muscle. Also, mineral profile differed between muscles with higher content of iron, significantly higher (P<0.001) content of zinc and lower content of calcium, magnesium and phosphorus (P<0.05) for ST muscle compared with RA. Therefore, supplementation with Cu modified deer meat characteristics, but it did not increase its concentration to toxic levels, making it a safe practice from this perspective. Despite the lower content of polyunsaturated FA, quality was better for RA than for ST muscle based on its higher content of protein with more essential/non-essential AA ratio and lower pH24 and cholesterol content.

Influence of sex and terminal sire line on performance and carcass and meat quality of Iberian pigs reared under intensive production systems.

A trial was conducted to study the effects of sex (castrated males; females) and terminal sire line (DD, Danish Duroc; SD, Spanish Duroc; RIB, Retinto Iberian) on performance and carcass and meat quality of pigs slaughtered at 145kg body weight. The female line was pure Iberian in all cases. Each treatment was replicated five times (six pigs). Females were more efficient, had less carcass and inter- and intramuscular fat and more trimmed primal cuts yield than castrated males. Duroc sired pigs grew faster and had better feed conversion and carcass quality than RIB sired pigs. Crossbreeds from SD had less carcass yield than crossbreeds from DD or RIB. We conclude that productive performance and primal cuts yield were higher for females than for castrated males. Danish Duroc sires are an alternative to Spanish Duroc and Retinto Iberian sires for production of heavy pigs destined for the dry cured industry.

Effect of gender and castration of females and slaughter weight on performance and carcass and meat quality of Iberian pigs reared under intensive management systems.

A total of 360 Iberian dam×Duroc sire pigs was used to study the influence of gender and castration of females (CM, castrated males; CF, castrated females; IF, intact females) and slaughter weight (SW; 145 and 156kg body weight, BW) on performance and carcass and meat quality. Each treatment was replicated four times (15 pigs). Intact females ate less, tended to have better feed conversion and had less carcass fat and more primal cuts yield than CM and CF. Pigs slaughtered at 156kg BW ate more feed and had worse feed conversion than pigs slaughtered at 145 BW. An increase in SW improved carcass yield, but tended to decrease trimmed ham yield. Intact females are an alternative to castrated females for intensive production of Iberian pigs. Also, the reduction in slaughter weight from 156 to 145kg BW is recommended for this type of production.

Productive performance and egg quality of brown egg-laying hens in the late phase of production as influenced by level and source of calcium in the diet.

A total of 1,152 Lohmann Brown laying hens were used to study the influence of level (3.5 and 4.0%) and source (coded FIN, COA, and OYS) of Ca in the diet on productive performance and egg quality from 58 to 73 wk of age. The FIN diet contained all the Ca carbonate as fine limestone (LIM). In the COA and OYS diets, 40% of the fine LIM was substituted with either coarse LIM or oyster shell. Each treatment was replicated 8 times (24 hens). Productive performance and egg quality traits were recorded every 4 wk, and tibia characteristics and shell quality traits were determined at 73 wk of age. An increase in Ca intake from 4.08 to 4.64 g/hen per day improved egg production (71.2 vs. 74.9%; P < 0.001), egg mass (49.0 vs. 51.4 g; P < 0.05), and feed conversion ratio (2.43 vs. 2.30 kg of feed/kg of egg; P < 0.001). In addition, an increase in Ca intake improved shell weight (9.98 vs. 10.20%; P < 0.05), shell thickness (0.342 vs. 0.351 mm; P < 0.01), and shell density (82.0 vs. 83.8 mg/cm2; P < 0.001). Calcium source had no effect on productive performance, tibia characteristics, or egg quality except for shell density, which was greater for hens fed COA than for hens fed FIN, with hens fed OYS being intermediate (81.9 vs. 84.0 vs. 82.7 mg/cm2, respectively; P < 0.05). It was concluded that Brown egg-laying hens in the late phase of production require more than 3.5% Ca in the diet (4.08 g of Ca/hen per day) and that the substitution of 40% of fine LIM with COA or OYS does not affect productive performance and has little impact on shell quality and tibia characteristics.

Effects of the levels of methionine, linoleic Acid, and added fat in the diet on productive performance and egg quality of brown laying hens in the late phase of production.

Two trials were conducted to study the effect of reducing the methionine (MET, 0.36 vs. 0.31%), linoleic acid (LIN, 1.60 vs. 1.12%), and supplemental fat (SFAT, 3.0 vs. 1.1%) content of isoenergetic diets on the productive performance and egg quality of brown laying hens late in the production cycle. The 8 treatments were arranged factorially (2 x 2 x 2), with 6 replicates of 20 hens per treatment (Hy-Line, from 59 to 70 wk of age in trial 1, and Lohmann, from 56 to 75 wk of age in trial 2). Except for SFAT content, dietary treatment had little effect on laying hen performance and egg quality. The only effect of a reduction in MET content on hen performance was the decrease in the percentage of large and extra large eggs (79.8 vs. 85.9%; P < 0.05) from 60 to 67 wk of age in trial 2. A decrease in the level of SFAT reduced egg production (79.3 vs. 77.0%; P < 0.05), egg weight (66.3 vs. 64.9 g; P < 0.001), egg mass (52.5 vs. 49.8 g/d; P < 0.001), feed conversion ratio (2.26 vs. 2.36 kg of feed/kg of eggs; P < 0.001), and percentage of extra large eggs (13.1 vs. 8.2%; P < 0.05) in trial 1, but no significant differences were detected in trial 2. Reducing the LIN content of the diet from 1.60 to 1.12% did not affect any trait in either of the 2 trials. We conclude that reducing the level of SFAT from 3.0 to 1.1% might decrease productivity and the percentage of extra large eggs. However, a reduction in the MET level from 0.36 to 0.31% and in LIN from 1.60 to 1.12% did not affect any trait in hens late in the production cycle. Eggshell quality was not affected by any of the dietary treatments.

Effects of Mn supplementation in late-gestating and lactating red deer (Cervus elaphus hispanicus) on milk production, milk composition, and calf growth.

This study describes the effects of Mn supplementation of 20 late-gestating and lactating Iberian red deer (Cervus elaphus hispanicus) females (hinds) fed a balanced diet on milk production and milk composition over the lactation period. Body weight of their calves at birth and at weaning was also evaluated. In addition, the effect of lactation stage was studied. For these purposes, 2 groups of hinds, one composed by 12 individuals (experimental) and the other by 8 individuals (control) were compared. Experimental hinds were s.c. injected weekly with Mn (2 mg Mn/kg BW) from day 140 of gestation until the end of lactation (week 18; forced weaning by physical separation). Control hinds were injected with a physiological saline solution with the same volume and at the same frequency as the experimental group. Serum Mn content of hinds was assessed just before the first Mn injection and at week 10 of lactation to assess whether the injected Mn increased Mn concentrations in blood. No differences were observed for BW of calves at birth but calves whose mothers were injected with Mn tended (P = 0.07) to have greater gain of BW from birth to weaning in proportion of BW at birth compared to calves from control hinds. In addition, supplementation with Mn increased (P ≤ 0.05) daily milk production by 10.2%, milk fat content by 11.2%, and total fat yield by 17.8%. Also, milk from hinds supplemented with Mn had more Ca (P < 0.001) and P (P < 0.05) than milk from control hinds. Manganese supplementation did not influence Mn serum content when blood was analyzed at week 10 of lactation, but increased the Mn content of milk by 18.3% (P < 0.001). Lactation stage affected (P < 0.001) fat, protein, lactose, and DM. Their contents increased as lactation proceeded, and protein was substituted by fat. Therefore, results suggest that Mn supplementation of hinds is recommended, even when they are fed a balanced diet, to increase milk production and the content of fat, Ca, P, and Mn of milk.

Does Cu supplementation affect the mechanical and structural properties and mineral content of red deer antler bone tissue?

The main factors affecting the mechanical (and other) properties of bone, including antler, are the proportions of ash (especially Ca and P) and collagen content. However, some trace minerals may also play more important roles than would be expected, given their low levels in bone and antler. One such trace mineral is Cu. Here, we studied the effects of Cu supplementation on the mechanical and structural characteristics, and mineral content of antlers from yearling and adult (4 years of age) red deer fed a balanced diet. Deer (n=35) of different ages (21 yearlings and 14 adults) were studied. A total of 18 stags (11 yearlings and 7 adults) were injected with Cu (0.83 mg Cu/kg BW) every 42 days, whereas the remaining 17 (10 yearlings and 7 adults) were injected with physiological saline solution (control group). The Cu content of serum was analysed at the beginning of the trial and 84 days after the first injection to assess whether the injected Cu was mobilized in blood. Also, the mechanical and structural properties of antlers and the mineral content in their cortical walls were examined at three (yearlings) or four (adults) points along the antler beam. The effect of Cu supplementation was different in yearlings and adults. In yearlings, supplementation increased the Cu content of serum by 28%, but did not affect antler properties. However, in adults, Cu supplementation increased the Cu content of serum by 38% and tended to increase the cortical thickness of antlers (P=0.06). Therefore, we conclude that, even in animals receiving balanced diets, supplementation with Cu could increase antler cortical thickness in adult deer, although not in yearlings. This may improve the trophy value of antlers, as well as having potential implications for bones in elderly humans, should Cu supplementation have similar effects on bones as those observed in antlers.

Nucleotide sequences of variable regions of an human anti-acetylcholine receptor autoantibody derived from a myasthenic patient.

Autoantibodies against the acetylcholine receptor (AChR) are involved in the neuromuscular dysfunction associated with myasthenia gravis (MG). We determined the nucleotide and the deduced amino acid sequences of the heavy and light chains for one human monoclonal anti-AChR autoantibody, derived from peripheral blood lymphocytes obtained from one MG patient. The heavy and light chain (VH and V lambda) genetic elements used in this autoantibody are respectively or closely related to HHG19 and Humlv117 germline structure. In addition, the expressed JH, J lambda and D segments differ from their described germline structures. This diversity could reflect allelic variation. The large number of differences found in VH, VL and D genetic elements when compared with their most closely related germline structures allowed us to conclude that we have characterized new VH and VL genes and we could not identify univocally somatic mutations. However, the analysed autoantibody, an IgG specific for the alpha chain of the AChR, revealed the presence of N addition segments on both sides of the D region and we may postulate that this antibody was the result of an antigen driven phenomenon.

Evidence for antigen driven selection in two monoclonal auto-antibodies derived from nonobese diabetic mice.

The nonobese diabetic (NOD) mouse is a model of human type I diabetes. This diabetes is due to massive infiltration of the pancreatic beta cell of islets by autoreactive T cells (insulitis) followed by the destruction of insulin-producing cells. Circulating autoantibodies are also detected, notably against glutamic acid decarboxylase, peripherin and insulin. Two monoclonal autoantibodies directed against insulin and peripherin were obtained by fusing NOD spleen and myeloma cells. We report here the nucleotide sequence of the genes encoding for the V regions of these two antibodies. Somatic mutations were identified by comparing the light chain nucleotide sequence of one of these autoantibodies with its germline counterpart precursor established from NOD mice after PCR gene amplification. The other one displays N additions on both sides of the D region. These results strongly suggest that both autoantibodies have undergone diversification, either N additions or somatic mutations, and therefore present structural features of antibodies derived from animals immunized against exogenous antigens.

Gastrointestinal autonomic nerve tumours and their separation from other gastrointestinal stromal tumours: an ultrastructural and immunohistochemical study of seven cases.

Gastrointestinal stromal tumours (GIST) represent a heterogeneous group whose classification frequently requires ultrastructural and immunohistochemical studies. In a retrospective study of the ultrastructural findings of 24 gastrointestinal stromal tumours, whose light microscopic study has yielded ambiguous results and in which accurate diagnosis had required ultrastructural support, seven were found to have the characteristics of gastrointestinal autonomic nerve (GAN) tumours. In all of them the diagnosis was based on the presence of dendritic processes with dense neuroendocrine granules. Immunohistochemically, the seven tumours were negative for smooth-muscle markers. All stained positively for vimentin. NSE, chromogranin, and synaptophysin were positive in most of them, while S-100 protein was positive only in two cases. We present the ultrastructural and immunohistochemical features of seven GANT against the background of the GISTs of our series. We conclude that GAN tumours cannot be diagnosed by light microscopy alone but this tumour group displays characteristic electron microscopic and immunohistochemical features and appears to represent a distinct type of GIST.