Transport of nutrients and electrolytes across the intestinal wall in pigs

In recent years, intestinal transport processes have been studied in detail regarding both, functional and structural aspects. For monosaccharides different systems have been demonstrated for apical uptake: this includes the high-affinity SGLT1 as a distinct d-glucose system and GLUT5 for fructose. Specifically in pigs a low affinity, high-capacity system for d-glucose and d-mannose with no preference for Na⁺ over K⁺ and a very low affinity system are suggested as further uptake systems. As in other species, basolateral extrusion is mediated by GLUT2. The distributions of monosaccharide transport along the gastrointestinal axis as well as the potential role of paracellular monosaccharide absorption have not yet been clarified.

Amino acids can principally be absorbed by the paracellular and transcellular pathway whereas transcellular transport can either be mediated by facilitated diffusion or secondary active Na⁺-coupled transport. This includes different transport systems for neutral, anionic and cationic acids. In addition, the presence of the di-/tripeptides transport system PEPT1 which depends on an inwardly directed H⁺-gradient has also been confirmed for the pig small intestine, its quantitative proportion is still under debate.

Short chain fatty acids (SCFA) are the major end products of microbial carbohydrate fermentation which occurs along the gastrointestinal tract with the highest production rates in the large intestines. At least two uptake mechanisms have to be assumed, i.e., non-ionic diffusion and anionic exchange via SCFA⁻/HCO₃⁻-exchange. Controversial views still exist to what extent SCFA are metabolized within the epithelial tissue.

Segmental differences between small and large intestines have been demonstrated for Na⁺ absorption. Whereas in the small intestines the major part of Na⁺ absorption is mediated by coupled nutrient transport systems, aldosterone sensitive Na⁺ channels and Na⁺/H⁺-exchange are the dominant mechanisms in the hindgut. For Cl⁻ paracellular transport and anionic Cl⁻/HCO₃⁻-exchange are the major absorptive mechanisms. Cl⁻ secretion is mediated by apical channels which may be activated by toxins of different origin. Different types of Cl⁻ channels have been identified, such as Cystic Fibrosis Transmembrane Regulator (CFTR), Ca-activated Cl⁻ channels (CLCA) and Outwardly Rectifying Cl⁻ Channels (ORCC). Whereas CFTR has clearly been shown for jejunal and colonic epithelial and goblet cells controversy still exists on the relevance of CLCA and ORCC in pigs.

For Ca²⁺ there is evidence that both recently published channels TRPV5 and TRPV6 are also expressed in pig intestinal tissues, however, this has not yet been shown on protein level. From several functional approaches it was demonstrated that phosphate uptake can be mediated by both, a Na⁺-dependent transcellular component and paracellularly. On a molecular basis it is uncertain whether the transport protein of transcellular mechanism belongs to the NaPi-IIb cotransporter family.     

Effects of VIP and GRF on the release of growth hormone in perifused bovine adenohypophysis

The effects of vasoactive intestinal polypeptide (VIP) and growth hormone releasing factor (GRF:hpGRF(1–29)-NH₂) on the release of growth hormone (GH) from anterior pituitaries from cows were examined by using an in vitro superfusion system. The pituitaries were excised randomly from cycling cows, dissected to obtain medial portions, and minced to obtain cubes with approximate dimensions of 1.5mm on a side. For each perifusion setup, 5 pieces of pituitary tissues were chambered and flushed with modified KRB solution saturated with 95% O₂-5% CO₂ at 38C. Perifusion with media containing 10⁻⁶ and 10⁻⁷M VIP for 30 min induced a significant release of GH during the treatments (P<0.05). VIP (10⁻⁸M) increased GH levels significantly (P<0.05), but to a minor degree. Perifusion with the media containing 10⁻⁶, 10⁻⁷ and 10⁻⁸M GRF for 30 min markedly increased the GH concentration and the effects continued up to 90 min after termination of the perifusion of the peptide (P<0.05, P<0.01). The GH releasing effects of GRF could be seen at doses as low as 10⁻¹¹M GRF (P<0.05, P<0.01).

These findings indicate that the GH releasing effect of VIP is less potent that that of GRF in cows.     

Studies on the pathogenesis of bovine ephemeral fever in experimental cattle III. Virological and biochemical data

In an attempt to define the nature of the response of cattle to ephemeral fever infection, a number of indicators of inflammation were monitored during clinical disease. The total Ca, Zn, Fe, Cu, glucose and phosphate in plasma, together with blood ammonia, were assayed relative to changes in the rectal temperature. Ca_T levels fluctuated markedly and hypocalcaemia occurred in 4 of 8 cattle. Plasma Zn and Fe values fell while plasma Cu levels rose markedly in all cattle. Mean levels of serum NH₃ of 20–30 μmol 1⁻¹ rose to a peak value of 56 μmoll⁻¹. Plasma glucose levels rose to a peak of 4.6 ± 0.5 mMl⁻¹ and the plasma phosphate levels fell from 2.4 ± 0.1 mMl⁻¹ to 1.17 ± 0.2 mMl⁻¹ during fever. Values of pCO₂ fell from a mean of 46.9 ± 3.6 mmHg to 36.4 ± 3.1 mmHg and coincided with a rise in pH. Virus was isolated 73 h (± 23) after inoculation and persisted until 130 h (± 21). The common role of these parameters in generalised inflammation and ephemeral fever is discussed.     

Physiological responses to water restriction in dry and lactating Awassi ewes

The effect of water restriction in Awassi ewes was assessed under two physiological conditions: lactating and dry, over a 3-week-period (August–September, 2002). Eight dry and eight lactating Awassi ewes (with their lambs) were assigned to one of two watering regimes: watered once every three days and daily watering, respectively. Weather data, body weight and rectal temperature were recorded. Venous blood was sampled on the watering day of the water-restricted group, and subjected to haematological, biochemical and electrolytes analysis. Ewes under water restriction lost more weight and had higher packed cell volume, haemoglobin, serum cholesterol, urea, creatinine, total protein and albumin as compared to daily-watered animals. However, lactation seemed to have no significant effect on these serum components. As for blood pH and electrolytes, Na⁺ and Cl⁻ were significantly higher under water restriction; whereas significantly lower K⁺ and Ca⁺⁺ but higher pH and Cl⁻ were observed in lactating animals. Finally, lower cortisol concentrations were observed in water-restricted animals, warranting further studies on the hormonal adaptation to long term water stress. It was concluded that water restriction induces similar changes in blood physiological indicators in dry and lactating Awassi ewes; while lactation beyond the first month post partum seems to mostly affect blood electrolytes.     

后海、后三里、大肠俞对大鼠便秘及胃肠运动神经递质的影响

后海（GV-1）、后三里（ST-36）、大肠俞（BL-25）是治疗动物便秘的常用穴位,本文旨在明确3个穴位对大鼠便秘的缓解作用及其对胃肠运动相关神经递质的调节作用,比较3个穴位作用途径的异同,探索便秘的优势穴位。选取SPF健康大鼠60只,随机分为6组。除对照A组外,其他5组大鼠每只每天灌服3 mg·（kg·d）^-1洛哌丁胺,建立便秘模型。5 d后,在便秘模型基础上,西药治疗C组灌服莫沙必利1.38 mg·（kg·d）^-1混悬液,D、E、F组分别针刺GV-1、ST-36、BL-25各30 min,连续6 d。检测血清5-羟色胺（5-HT）、生长抑素（SS）、血管活性肠肽（VIP）、P物质（SP）,结肠组织中5-羟色胺3受体（5-HT₃R）mRNA、5-羟色胺4受体（5-HT₄R）mRNA相对表达量及结肠组织结构。结果显示：造模5 d后,模型B组大鼠出现典型便秘症状,血清中5-HT、SS含量显著增加,VIP、SP显著下降,结肠组织5-HT₃R和5-HT₄R mRNA显著降低。西药治疗组和3个穴位组大鼠的便秘症状均显著缓解,首粒粪便时间缩短,粪便含水量增加,粪便长度增加。另外,针刺GV-1使血清中SS含量极显著降低36.34%（P<0.01）,5-HT含量显著下降。针刺双侧ST-36下调血清中5-HT含量23.93%,显著增加结肠组织5-HT₃R、5-HT₄R mRNA相对表达量（P<0.01）。针刺双侧BL-25,血清中VIP、SP含量分别提高27.48%、22.75%,显著降低5-HT含量。另外针刺3个穴位均可增加结肠杯状细胞表达。GV-1、ST-36、BL-25缓解便秘的途径有一定差异,但均能缓解大鼠便秘,改善胃肠道运动,调节胃肠道神经递质的含量,其中GV-1对便秘的改善作用最为显著。     

5-羟色胺影响肠黏膜上皮细胞屏障的研究进展

5-羟色胺（5-hydroxytryptamine,5-HT）作为一种重要的神经递质和胞间信使,具有多种生物学功能。90%以上的5-HT分布在胃肠系统中,在胃肠系统中发挥重要的生理学功能,许多肠道疾病的发生与肠道中5-HT的表达量变化有关。5-HT影响肠黏膜上皮细胞的增殖和维持,同时升高肠黏膜通透性,间接促进杯状细胞黏液的分泌。其作用机制可能是5-HT刺激传入神经元,激活血管活性肠肽、乙酰胆碱的分泌,间接引起腹泻。作者对5-HT引起腹泻的原因,以及对肠黏膜上皮细胞生物学功能的影响作一综述,以期对腹泻等肠道性疾病诊疗具有一定指导作用。     

硒、锌元素配施对紫花苜蓿产量、植株体内硒锌积累和氨基酸含量的影响

在严重缺硒缺锌的石灰性土壤中,采用锌、硒两因素三水平(纯锌施用量分别0、0.10、0.40 g·kg^-1,纯硒施用量 0、0.30、1.80 mg·kg^-1)完全设计盆栽基施的方法,以紫花苜蓿为对象,研究了硒、锌元素配施对紫花苜蓿产量、硒锌积累量和氨基酸含量与组成的影响,并采用氨基酸评分(AAS)、化学评分(CS)和必需氨基酸指数(EAAI)对紫花苜蓿氨基酸组成特点进行综合评价,从而探讨硒、锌复合微肥对紫花苜蓿营养价值的影响。结果表明,除单施低锌处理(0.10 g·kg^-1)外,其他处理均显著提高苜蓿干草产量,硒是提高产量的主要因素,锌对提高苜蓿干草产量有促进作用,两者配合效果最好;单施硒或硒锌配施均极显著提高了苜蓿硒含量,硒锌间表现为协同作用;单施锌极显著提高了苜蓿锌含量,配施适量硒促进了紫花苜蓿对锌的吸收,但过量施硒不利于苜蓿对锌的吸收。各处理紫花苜蓿总氨基酸含量(T)在15.88~18.89 g·100 g^-1(以干重计),以低硒低锌处理(Se₁Zn₁)含量最高,高硒高锌处理(Se₂Zn₂)含量最低。各处理紫花苜蓿必需氨基酸含量(E)在5.11~6.45 g·100 g^-1,以低硒高锌处理(Se₁Zn₂)含量最高,Se₂Zn₂处理含量最低;低硒(Se₁Zn₀)、高硒低锌(Se₂Zn₁)、高锌(Se₀Zn₂)与低硒高锌(Se₁Zn₂)等处理紫花苜蓿氨基酸E/T值接近40%。所有处理紫花苜蓿蛋白质的第一限制氨基酸均为含硫氨基酸(蛋氨酸+胱氨酸)。可见,在严重缺硒和锌的石灰性土壤中,施锌量0.1~0.4 g·kg^-1,施硒量0.3 mg·kg^-1不仅能提高紫花苜蓿产量,而且能改善其氨基酸组成和品质。     

Effects of dietary rhIGF-I in neonatal calves on the appearance of glucose, insulin, D-xylose, globulins and γ-glutamyl transferase in blood

Cow colostrum is rich in insulin-like growth factors IGF-I and II, thus the dietary effects of recombinant human (rh) rhIGF-I on the newborn were of interest. The objective of this study was to examine the effects of dietary IGF-I upon selected blood components and gut absorptive development. Calves were blocked by birth weight and fed two times per d for a total of four times with the initial restricted diet. The initial feeding was 1.5 l and the remaining three feedings were at 2 1 with one of three experimental diets: 1) milk replacer plus isolated colostrum derived globulins (MR⁻), 2) same as 1 above plus 750 ng/ml rhIGF-I (MR⁺), 3) pooled cow colostrum (COL). Thereafter, all animals received only milk replacer at 5% of body weight (BW)/feeding two times per d with only treatment 2 having continued addition of 750 ng/ml rhIGF-I until experimental completion at 6 to 7 d after birth. At feeding three, animals were fed D-xylose (0.5 g/kg BW) and 5,000 U of bovine kidney membrane γ-glutamyl transferase as indicators of gut absorptive capacity. Colostrum-fed animals received 5,000 U of natural occurring γ-glutamyl transferase activity in the 1.5 l first feeding. Blood samples taken over time were collected and saved as frozen plasma. All diets were analyzed for nutrient composition and endogenous levels of test hormones. Colostrum fed calves had greater globulin concentration (P < 0.01) than MR⁻ or MR⁺ fed calves. Recombinant hIGF-I feeding had no effect (MR⁻ vs. MR⁺) upon total protein, albumin or globulin blood levels. Absorption of colostrum γ-glutamyl transferase at first feeding resulted in a peak total blood U of 4.5% of that fed. Although enzyme absorption was greatly reduced by the third feeding (0.5% of total fed), MR⁺ fed calves exhibited no significant difference in enzyme absorption when compared with the controls (MR⁻ vs. MR⁺). However, pharmacokinetic analysis of D-xylose absorption at the third feeding showed diet effect upon absorption of D-xylose. Dietary rhIGF-I may change development or activity of sugar transporters and also may alter absorption of macromolecules (closure) in neonatal calves.     

Effect of a nonsteroidal aromatase inhibitor on in vitro and in vivo secretion of estradiol and on the estrous cycle in ewes.

Three experiments were performed to study effects of decreased concentrations of estradiol-17β (E₂) on lifespan and function of ensuing ovine corpora lutea (CL). In experiment 1, 52 follicles were collected from 10 ewes and placed into individual culture with 0 or .01 μCi ³H-androstenedione (10 ng; ³H-A) and 0, 10⁻¹¹, 10⁻⁹, 10⁻⁷, or 10⁻⁵ M of a nonsteroidal aromatase inhibitor, CGS16949A (CGS). Concentrations of E₂ secreted into the medium, and synthesis of estrogens as estimated by formation of ³H-water from ³H-A were decreased by 10⁻⁵ and 10⁻⁷ (P<.01), but not 10⁻⁹ or 10⁻¹¹ M CGS. In experiment 2, luteolysis was induced in 24 ewes by injection of PGF₂ on days 5 to 10 of the estrous cycle (0 hr). Ewes received 0, 0.5, 1.0, 2.0 or 4.0 mg CGS per kg BW i.v. at −12, 0, 12 and 24 hr, and an ovulatory dose of hCG at 36 hr. Jugular (P<.001) and vena caval (P<.001) concentrations of E₂ were decreased by CGS at all doses tested for 8 to 10 hr, but had returned to levels similar to control ewes by the time of the next injection. Concentrations of E₂ around the time of the LH surge were similar in control and treated ewes. During the subsequent luteal phase, concentrations of progesterone (P₄) were similar in control and treated ewes. Thus, transient decreases in E₂ during the follicular phase were not deleterious to the subsequent luteal phase. In experiment 3, luteolysis was induced in 18 ewes by injection of PGF₂ on days 6 or 7 (0 hr) of the estrous cycle. Ewes received 0 or 1 mg CGS per kg BW i.v. every 8 hr from 0 to 40 hr. Ovulation was induced with hCG at 36 hr. CGS reduced jugular (P<.001) and vena caval (P<.001) concentrations of E₂, prevented an endogenous surge of LH (P<.05) and increased (P<.001) concentrations of FSH. All ewes had ovulated a marked follicle by 72 hr, but onset of the luteal phase, as assessed by concentrations of P₄, was delayed (P<.01) in ewes receiving CGS. Delayed luteal phases were not solely attributable to the presence of new CL or to luteinization of follicular cysts. When data were aligned according to the day ewes were observed in estrus, profiles of P₄ did not differ with treatment. Therefore, normal luteal function ensued following estrus whether or not ewes re-ovulated. In conclusion, decreased secretion of E₂ by the preovulatory follicle was not involved in the ontogeny of CL of short lifespan or subnormal function. Instead, adequate production of E₂ or precisely timed E₂ secretion may be required during follicular development for subsequent functional luteinization.     

Determination of the source of increased serotonin (5-HT) concentrations in blood and peritoneal fluid of colic horses with compromised bowel

REASONS FOR PERFORMING STUDY: Increased plasma (5-HT) concentrations are reported in horses predisposed to develop laminitis and after i.v. infusion of endotoxins. In the equine jejunum contractile 5-HT1A-like receptors show tachyphylaxia upon prolonged activation with 5-HT. Therefore, increased systemic 5-HT release in colic horses could play a possible role in the pathophysiology of ileus. OBJECTIVE: To investigate possible increased systemic release of 5-HT in colic horses with compromised bowel and to identify the source of 5-HT overload. METHODS: Concentrations of 5-HT were determined in plasma and peritoneal fluid (PF) of healthy horses (n = 10), strangulating small intestinal colic horses (n = 18), nonsurgical colic horses (n = 10) and cryptorchid stallions (n = 6). It was attempted to identify the source of 5-HT overload by comparing the blood and PF 5-HT concentrations within horses and by assessing the in vivo platelet activation through determination of the beta-thromboglobulin (beta-TG)/platelet factor 4 (PF4) ratio. RESULTS: All horses in the strangulating small intestinal colic group had plasma (P = 0.006) and PF (P = 0.01) 5-HT concentrations above those found in the control group. Plasma beta-TG/PF4 ratio in these horses exceeded 2 in all cases, indicating in vivo platelet activation. Concentrations of 5-HT in PF of colic horses with compromised bowel were significantly lower than the corresponding plasma concentrations (P = 0.005). Potential relevance: In horses with compromised bowel, significant amounts of 5-HT can be released into the systemic circulation, through massive release of platelet-stored 5-HT. 5-HT is a very potent proinflammatory, vasoconstrictive and immunomodulatory agent. In view of the rapid and prolonged tachyphylaxia, shown for the jejunal 5-HT1A-like receptors, this increased systemic 5-HT release could play a role in the pathophysiology of ileus in horses.     

Modulation of growth hormone-releasing factor-induced release of growth hormone from bovine pituitary cells

Growth hormone (GH)-releasing factor (GRF) at concentrations of 10−¹² through 10−⁷M for 6 hr linearly increased GH release (b₁ = 10.4 ± .3) from bovine anterior pituitary cells in culture. Maximum release of GH (262% above controls) occurred at 10−⁷M GRF. In contrast, GH release-inhibiting factor (SRIF) at 10−¹² through 10−⁵M had no effect on basal concentrations of GH. In a second experiment, as the proportion of SRIF relative to GRF increased. SRIF suppression of GRF-induced GH release from anterior pituitary cells increased. In a third experiment, anterior pituitary cells cultured in media containing fetal calf serum (FCS) were treated with cortisol (0 or 10 ng/ml media) for 24 hr before exposure to 10−¹³ through 10−⁷M GRF. GRF linearly increased GH secretion (b₁ = 7.4 ± .3) and cortisol augmented this response (b₁ = 10.5 ± .6). However, when cells were cultured in media containing dextran-charcoal treated FCS, cortisol did not alter GRF-induced GH release. Our results demonstrate that GH response of bovine anterior pituitary cells to GRF was modulated negatively by SRIF. However, augmentation of GRF-induced GH release by cortisol was evident only when cells were cultured in media supplemented with untreated FCS.     

Perforin expression can define CD8 positive lymphocyte subsets in pigs allowing phenotypic and functional analysis of natural killer, cytotoxic T, natural killer T and MHC un-restricted cytotoxic T-cells

In this study we have used the expression of perforin to characterize subsets of porcine cytotoxic lymphocytes. Perforin positive lymphocytes expressed both CD2 and CD8, most were small dense lymphocytes (SDL) and up to 90% were CD3 negative. However, the numbers of perforin positive T-cells increased with the age of the animal and their populations increased after specific antigen stimulation in vitro. The remaining perforin positive lymphocytes were large and granular and contained more CD3⁺CD5⁺CD6⁺ T-cells (−40%) of which a substantial proportion also co-expressed CD4. Perforin was expressed in subpopulations of both CD8 and CD8β lymphocytes, but was not expressed in γδ T-cells or monocyte/macrophages. The perforin positive CD3⁻ subset was phenotypically homogeneous and defined as CD5⁻CD6⁻CD8β⁻CD16⁺CD11b⁺. This population had NK activity and expressed mRNA for the NK receptor NKG2D, and adaptors DAP10 and DAP12. Perforin positive T-cells (CD3⁺) could be divided into at least three subsets. The first subset was CD4⁻CD5⁺CD6⁺CD11b⁻CD16⁻ most were small dense lymphocytes with cytotoxic T-cell activity but not all expressed CD8β. The second subset was mainly observed in the large granular lymphocytes. Their phenotype was CD4⁺CD5⁺CD6⁺CD8β⁺CD16⁻CD11b⁻ and also showed functional CTL activity. Thus not all of double positive T-cells are memory helper T-cells. The third subset did not express the T-cell co-receptor CD6, but up to half of them expressed another T-cell co-receptor CD5. The majority of this subset expressed CD11b and CD16, thus the third perforin positive T-cell subset was CD3⁺CD4⁻CD5⁺CD6⁻CD8β^±CD11b⁺CD16⁺, and possessed MHC-unrestricted cytotoxicity and LAK activity.     

Effects of a second mutant allele (V199I) at the PRKAG3 (RN) locus on carcass composition in pigs

The effect of a second mutant allele (V199I, here denoted rn*) at the PRKAG3 (RN) locus on carcass composition was determined in 334 pigs, entire males and females, from crosses between Swedish Hampshire (H) and Finnish Landrace (L) (H × LH; LH × H; LH × LH). Pigs were classified according to DNA test into the following PRKAG3 genotypes: RN⁻/RN⁻ (23%), RN⁻/rn⁺ (24%), RN⁻/rn* (33%), rn⁺/rn⁺ (8%), rn⁺/rn* (9%) and rn*/rn* (2%). The pigs were slaughtered at a commercial slaughterhouse and assessed 24 h postmortem. Right sides were fabricated into primary wholesale cuts, then further processed into defatted hams and loins, and a subset of hams (n = 122) was dissected into the five major individual muscles. The genotype frequencies for the subsample were RN⁻/RN⁻ (27%), RN⁻/rn⁺ (20%), RN⁻/rn* (35%), rn⁺/rn⁺ (9%), rn⁺/rn* (8%) and rn*/rn* (1%). Weights were recorded for meat and bone in ham and loin, fat in ham, back and shoulder and the individual dissected muscles. The genotype effect was significant (P < 0.05) for estimated lean meat content and the proportions of meat and bone and fat in ham and loin (of carcass weight). Also, the content of meat and bone in ham and loin, in proportion of whole ham and loin, respectively, differed significantly (P < 0.01) between genotypes. Estimated lean meat content was highest for RN⁻/RN⁻ (63.0%) and RN⁻/rn⁺ (63.1%) and lowest in the combined group rn*/⁻ (rn⁺/rn* and rn*/rn*, 61.7%); RN⁻/rn* (62.5%) and rn⁺/rn⁺(62.1%) were intermediate. The same results were found for meat and bone in ham and loin, as a proportion of whole ham and loin, respectively. RN⁻/RN⁻ and RN⁻/rn⁺ did not differ in any trait; however, they produced carcasses with the lowest proportions of fat within loin and the major wholesale cuts (ham, loin and shoulder). The carcass percentage of meat and bone in ham was higher in the three RN⁻/ genotypes (RN⁻/RN⁻, RN⁻/rn⁺ and RN⁻/rn*, P < 0.05) than in the rn*/⁻ group, whereas rn⁺/rn⁺ did not (P > 0.05) differ from any of the other genotypes. RN⁻/rn⁺ and RN⁻/rn* had higher (P < 0.05) proportion of meat and bone in loin compared to the rn*/⁻ group. We conclude that the second mutant allele found at the PRKAG3 (RN) locus, rn*, decreased the lean meat content compared with the two other alleles (RN⁻, rn⁺). The RN⁻/RN⁻ and RN⁻/rn⁺ genotypes were leanest, followed by RN⁻/rn* and rn⁺/rn⁺, and rn⁺/rn* and rn*/rn* were the fattest.     

Dose effect of human growth hormone-releasing factor and thyrotropin-releasing factor on hormone concentrations in lactating dairy cows

Two experiments were conducted to study the effects of growth hormone-releasing factor (GRF) and thyrotropin-releasing factor (TRF) administration on hormone concentrations in dairy cows. In the first trial, 12 cows were used on 5 consecutive days to determine the effect of four sc doses of GRF (0, 1.1, 3.3 and 10 μg•kg⁻¹ BW) and three sc doses of TRF (0, 1.1 and 3.3 μg•kg⁻¹ BW) combined in a factorial arrangement. GRF and TRF acted in synergy (P = .02) on serum growth hormone (GH) concentration even at the lowest dose tested and GH response to the two releasing factors was higher than the maximal response observed with each factor alone. TRF increased (P<.01) prolactin (Prl), thyrotropin (TSH), triiodothyronine (T₃) and thyroxine (T₄) concentrations similarly at the 1.1 and 3.3 μg•kg⁻¹ doses and GRF did not interact (P>.40) with TRF on the release of these hormones. In the second trial, the effect of GRF (3.3 μg•kg⁻¹ BW, sc) and TRF (1.1 μg•kg⁻¹ BW, sc) was tested at three stages (18, 72 and 210 days) of lactation on serum Prl and TSH concentrations. Eighteen cows (n = 6 per stage of lactation) were used in two replicates of a 3 × 3 latin square. The TRF and GRF-TRF treatments were equipotent (P>.05) in increasing Prl and TSH concentrations. Prl and TSH responses were similar (P>.40) throughout lactation. In summary, GRF at doses ranging from 1.1 to 10.0 μg•kg⁻¹ and TRF at doses ranging from 1.1 to 3.3 μg•kg⁻¹ act in synergy on GH release and do not interact on Prl, TSH, T₃ and T₄ concentrations in dairy cows. Furthermore, Prl and TSH response to TRF are not affected by stage of lactation.     

Effects of indomethacin on Salmonella typhimurium- and cholera toxin-induced fluid accumulation in the porcine small intestine

The effect of the cyclooxygenase and prostaglandin E2 (PGE2) synthesis inhibitor, indomethacin, on the secretory responses induced by Salmonella serotype Typhimurium (ST) and cholera toxin (CT), in the porcine small intestine was investigated. ST (10(10) colony-forming units) and CT (56 micrograms) were instilled in tied-off intestinal loops in young anaesthetized pigs receiving intravenous indomethacin in a total dose of 7.5 mg/kg, or saline. The accumulated fluid in the loops and the luminal content of endogenous secretagogues PGE2 and 5-hydroxytryptamine (5-HT) were measured. ST induced fluid accumulation in the jejunum, whereas CT induced fluid accumulation in the jejunum and ileum. Indomethacin had no effect on the secretory responses. Indomethacin had a significant effect on the luminal content of PGE2 in jejunal ST and CT loops, whereas no effect of indomethacin was observed on the luminal content of 5-HT in ST and CT loops. In ST and CT loops, an increased content of PGE2 and 5-HT compared with test loops infused with Ringer's solution was observed. These results indicate that the porcine jejunal secretory response to ST and CT does not involve prostaglandins although indomethacin has an influence on the luminal release of PGE2 but not of 5-HT.     

The C bicarbonate dilution technique to determine energy expenditure in young bulls validated by indirect calorimetry

We explored the applicability of the ¹³C bicarbonate dilution technique for determination of energy expenditure (EE) in young bulls in comparison to whole body indirect calorimetry (IC). Twelve bulls of a F2 German Holstein x Charolais cross (4.5 months, 332 ± 16 kg BM) received a diet providing 1000 kJ ME d^− 1 kg BM^− 0.75 and 4.3 g crude protein d^− 1 kg BM^− 0.75. Bulls were housed in respiration chambers and received an intravenous bolus of NaH¹³CO₃ (A: 3 μmol kg BM^− 1 (n = 2), B: 7 μmol kg BM^− 1 (n = 4), C: 17.5 μmol kg BM^− 1 (n = 6), 99 at.% ¹³C) into the jugular vein to measure EE. Simultaneously, EE was determined by IC. After the ¹³C administration blood samples and breath gas were collected from the animals in the respiration chamber during a 24-h period (7.00–7.00 h). The recovery of ¹³C in breath CO₂ (% of ¹³C dose) was irrespective of NaH¹³CO₃ dose (A: 69.7 ± 2.7%, B: 70.5 ± 4.5%, C: 75.0 ± 4.9%; P > 0.05). Only small amounts of ¹³C were excreted in urine (3.4 ± 2.6%) and feces (2.0 ± 1.3%). The EE determined by the ¹³C bicarbonate method using breath and blood ¹³C recovery rates as correction factors was not different from that measured by IC (816 ± 81 [blood] or 827 ± 101 [breath] vs. 820 ± 90 kJ d^− 1 kg BM^− 0.75). Bland–Altman analysis showed a 95% confidence interval for EE of ± 99 and ± 109 kJ d^− 1 kg BM^− 0.75 based on blood and breath ¹³C recovery, respectively. In conclusion, the ¹³C bicarbonate dilution method is appropriate to obtain reliable estimates of EE in young bulls using blood CO₂ or breath CO₂ under standardized experimental conditions, i.e. in the fasting state.     

Observations on production of hemolysin, heat-labile enterotoxin and antimicrobial drug resistance among enterotoxigenic Escherichia coli from pigs.

A total of 52 isolates of Escherichia coli belonging to enterotoxigenic serotypes from piglets with diarrhea were examined for hemolysis, production of cholera-like enterotoxin (LT) and susceptibility to 10 antimicrobial drugs. A strong association between production of LT and hemolysis was seen. Ninety percent of 29 hemolylic isolates were LT⁺ whereas 100% of 23 nonhemolytic isolates were LT⁻ in a commercial latex agglutination assay. Antimicrobial susceptibility tests employing disc diffusion showed that resistance to trimethoprim-sulfamethoxazole (TMS), neomycin and tetracycline was significantly less among LT⁺ isolates compared to LT⁻ ones. Enrofloxacin was the only antimicrobial drug to which all the 52 isolates were susceptible.     

玉屏风多糖对小鼠派氏结形态结构及其T细胞亚群的影响

通过研究玉屏风多糖（YPF-P）对小鼠派氏结（Peyer's patches,PPs）形态结构及其T细胞亚群的影响,探讨其对小鼠肠黏膜的免疫调节作用。选取96只SPF小鼠随机分为6组,分别为空白对照组（0.2 mL生理盐水）、YPF-P阳性对照组（YPF-P 200 mg/kg）、环磷酰胺（cyclophosvnamide,Cy）免疫抑制组（80 mg/kg Cy）及YPF-P低、中、高剂量组（100、200、400 mg/kg YPF-P）,饲喂1周,取小肠PPs,常规切片HE染色后应用图像分析技术检测PPs形态结构的变化;体外培养小鼠PPs淋巴细胞,采用流式细胞术研究PPs中T淋巴细胞亚群的变化。结果表明,YPF-P对小鼠小肠PPs的生长发育具有促进作用,Cy可极显著降低小鼠小肠PPs面积、小肠纵切面面积及PPs面积和小肠纵切面面积的比值（P < 0.01）,低、中、高剂量YPF-P可在一定程度上缓解Cy对小鼠小肠的损伤作用。同时,YPF-P可显著或极显著提高PPs中CD3⁺、CD4⁺ T淋巴细胞及CD4⁺/CD8⁺（P < 0.05; P < 0.01）。结果显示,YPF-P能提高Cy诱导的免疫抑制小鼠的肠黏膜免疫功能,并能促进PPs中相关T淋巴细胞增殖,对小鼠肠黏膜功能具有增强作用。     

Effects of Recombinant E. coli Expressing Heat-stable Enterotoxin on Intestinal Morphology and Antioxidant Capacity in Seven Days old Piglets

This experiment was conducted to investigate the effects of recombinant E.coli expressing heat-stable enterotoxin(STa) on intestinal absorption and barrier function, intestinal morphology and antioxidant capacity of 7 days old piglets. Twenty-four 7 days old piglets were allotted to four treatments:control group (artificial milk), STa group (artificial milk +2×10⁹ CFU E.coli LMG194-pBAD-STa), LMG194 group (artificial milk +2×10⁹ CFU E.coli LMG194), and K88 group (artificial milk +2×10⁹ CFU E.coli K88).The pigs were treated with E.coli on the 5th day and slaughtered on the 7th day. The results showed that, compared with the control group, villus height in jejunum, ileum and duodenum, crypt depth and villus height/crypt depth in duodenum, and small intestine villi surface area were significantly decreased in STa group (P<0.05). The activities of catalase (CAT) in ileum and colon, total nitric oxide synthase (TNOS) in serum, ileum, jejunum and colon, inducible nitric oxide synthase (iNOS) in colon were significantly decreased in STa group (P<0.05). STa group also had a higher levels of malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) in serum (P<0.05). These results suggested that recombinant E.coli expressing STa could lead to intestinal injury and oxidative stress of 7 days old piglets.     

Effects of age and zinc supplementation on transport properties in the jejunum of piglets

Zinc is effective in the prevention and treatment of post‐weaning diarrhoea and in promoting piglet growth. Its effects on the absorption of nutrients and the secretory capacity of the intestinal epithelium are controversial. We investigated the effects of age, dietary pharmacological zinc supplementation and acute zinc exposure in vitro on small‐intestinal transport properties of weaned piglets. We further examined whether the effect of zinc on secretory responses depended on the pathway by which chloride secretion is activated. A total of 96 piglets were weaned at 26 days of age and allocated to diets containing three different levels of zinc oxide (50, 150 and 2500 ppm). At the age of 32, 39, 46 and 53 days, piglets were killed, and isolated epithelia from the mid‐jejunum were used for intestinal transport studies in conventional Ussing chambers, with 23 μm ZnSO₄ being added to the serosal side for testing acute effects. Absorptive transport was stimulated by mucosal addition of d ‐glucose or l ‐glutamine. Secretion was activated by serosal addition of prostaglandin E₂, carbachol or by mucosal application of Escherichia coli heat‐stable enterotoxin (St_p). Jejunal transport properties showed significant age‐dependent alterations (p < 0.03). Both absorptive and secretory responses were highest in the youngest piglets (32 d). The dietary zinc supplementation had no significant influence on jejunal absorptive and secretory responses. However, the pre‐treatment of epithelia with ZnSO₄ in vitro led to a small but significant decrease in both absorptive and secretory capacities (p < 0.05), with an exception for carbachol (p = 0.07). The results showed that, in piglets, chronic supplementation with zinc did not sustainably influence the jejunal transport properties in the post‐weaning phase. Because transport properties are influenced by the addition of zinc in vitro, we suggest that possible epithelial effects of zinc depend on the acute presence of this ion.