Effect of progressive cachectic parasitism and growth hormone treatment on hepatic 5'-deiodinase activity in calves

Thyroid status is compromised in a variety of acute and chronic infections. Conversion of thyroxine (T₄) into the metabolically active hormone, triiodothyronine (T₃), is catalyzed by 5′-deiodinase (5′D) mainly in extrathyroidal tissues. The objective of this study was to examine the effect of protozoan parasitic infection (Sarcocystis cruzi) on hepatic 5′D (type I) activity and plasma concentrations of T₃ and T₄ in placebo- or bovine GH (bGH)-injected calves. Holstein bull calves (127.5±2.0 kg BW) were assigned to control (C, ad libitum fed), infected (I, 250,000 S. cruzi sporocysts per os, ad libitum fed), and pair-fed (PF, non-infected, fed to intake of I treatment) groups placebo-injected, and three similar groups injected daily with pituitary-derived bGH (USDA-B-1, 0.1 mg/kg, i.m.) designated as C_GH, I_GH and PF_GH. GH injections were initiated on day 20 post-infection (PI), 3–4 days prior to the onset of clinical signs of the acute phase response (APR), and were continued to day 56 PI at which time calves were euthanized for liver collection. Blood samples were collected on day 0, 28, and 55 PI. Alterations in nutritional intake did not affect type I 5′D in liver. Treatment with bGH increased (P<0.05) 5′D activity in C (24.6%) and PF (25.5%) but not in I calves. Compared to PF calves, infection with S. cruzi reduced 5′D activity 25% (P<0.05) and 47.8% (P<0.01) in placebo- and bGH-injected calves, respectively. Neither nutrition nor bGH treatment significantly affected plasma concentrations of T₄ and T₃ on day 28 and 55 PI. However, plasma thyroid hormones were reduced by infection. On day 28 PI, the average plasma concentrations of T₃ and T₄ were reduced in infected calves (I and I_GH) 36.4% (P<0.01) and 29.4% (P<0.05), respectively, compared to pair-fed calves (PF and PF_GH). On day 55 PI, plasma T₃ still remained lower (23.7%, P<0.01 versus PF) in infected calves while plasma T₄ returned to control values. The data suggest that parasitic infection in growing calves inhibits both thyroidal secretion and extrathyroidal T₄ to T₃ conversion during the APR. After recovery from the APR, thyroidal secretion returns to normal but basal and bGH-stimulated generation of T₃ in liver remains impaired.     

Perturbed metabolism and hormonal profiles in calves infected with Sarcocystis cruzi

Plasma concentrations of growth hormone (GH), thyroid stimulating hormone (TSH), insulin (IN), thyroxine (T₄), and triiodothyronine (T₃) in addition to metabolic parameters [N balance (NB), urinary 3-methylhistidine (TMH), urinary creatinine (CR), and urinary hydroxyproline (HP)] were measured in 4-mo-old Holstein steers divided equally among groups infected with Sarcocystis (I), noninfected ad libitum fed (C), and noninfected pair fed to I (PF) (7 steers per treatment). Effects of infection on these parameters beyond those attributable to altered dietary intake were determined using orthogonal contrasts (effect of intake, C vs I + PF; effect of infection, PF vs I). NB was higher in C than I and PF (P<.05) and lower in I than PF (P<.02). Hydroxyproline and CR were influenced by intake (P<.05) and HP excretion was reduced in association with infection (P<.05). Reduced intake was associated with lowered mean basal plasma concentrations of GH, IN, T₃ and T₄ (P<.05). Infection further reduced (P<.001) plasma T₃ concentration.

Triiodothyronine and T₄ responses following an intravenous bolus of thyrotropin releasing hormone (TRH) were measured. The magnitude of the responses in I and PF were lower than those observed in C (P<.05). Plasma T₃ responses were further reduced in association with infection (P<.05). Insulin responses to intravenous arginine infusion (ARG) were also low in association with reduced intake. Growth hormone responses to TRH or ARG were affected by the level of feed intake only. These data suggest that hormonal perturbations associated with the insult of infection further compromise metabolism and the direction of nutrient partitioning that would ordinarily be associated with developmental growth in young steers beyond those responses anticipated from solely the reduction of feed intake.     

Serological and bacteriological observations on experimental infection with Salmonella hadar in chickens

Over the past 3 years the frequency of Salmonella hadar infections has increased in Belgium in both poultry and humans. Therefore, the course of infection with S. hadar in poultry was investigated. One day-old and 4 week-old specific pathogen-free chickens were orally infected with one of two S. hadar strains, SH1 or SH2. Mortality was 6% (SH1) and 17% (SH2) in birds infected at 1 day-old. Chickens infected at 1 day-old with SH2 showed a mild diarrhoea. The S. hadar faecal excretion in birds infected at 1 day-old remained high throughout the experiment until 12 weeks post-inoculation (pi). Faecal excretion was lower in older birds. Antibodies to S. hadar were observed from 2 weeks pi (SH2, infected at 1 day-old) or 4 weeks pi (SH1, both groups; SH2, chickens infected at 4 weeks of age). The percentage of chickens with antibodies was higher after infection at 1 day-old than after infection at 4 weeks of age. In a second experiment 1 day-old chicks were infected with SH1 and autopsied at regular intervals until 42 days pi. SH1 was isolated from the caeca from 3 h pi onwards and from the liver and spleen from 18 h until 14 days pi. Serous typhlitis and omphalitis were the main lesions. The number of macrophages in the lamina propria of the caecal tonsils was slightly increased from 18 h until 2 weeks pi. In the liver, inflammation was observed in the portal triads and in the sinusoids. This study indicates that infections with S. hadar lead to intense colonisation of the gut and extensive faecal shedding. It may also cause invasive infections in 1 day-old chickens.     

CHANGES IN SOMATOTROPIC AXIS RESPONSE AND BODY COMPOSITION DURING GROWTH HORMONE ADMINISTRATION IN PROGRESSIVE CACHECTIC PARASITISM

A multistage protozoan parasitic disease was used as a cachexia model to study the effects of daily administration of bovine growth hormone (GH) on endocrine and body composition changes of young calves from the onset of the acute phase response (APR). Male calves averaging 127.5 ± 2.0 kg body weight were assigned to control, ad libitum fed, noninfected (C); ad libitum fed, infected (250,000 oocysts Sarcocystis cruzi, per os, I); noninfected, pair-fed (PF) to matched I-treatment calves and these respective same treatments in calves injected daily with GH (USDA-bGH-B1), 12.5 mg/calf/day, im) designated as C_GH, I_GH and PF_GH. GH injections were initiated on Day 20 postinfection (PI), 3 to 4 d before the onset of clinical signs of APR, and continued to Day 56 PI, at which time animals were euthanized for tissue collections. Abrupt increases in rectal temperature commensurate with up to 70% reduction in voluntary feed intake were observed in I and I_GH beginning 23–25 d PI. For the trial period between Days 20 and 56 PI, average daily carcass protein gains were 123, 52, 109, 124, 48, and 67 g/d and average daily carcass fat gains were 85, 11, 43, 71, −23, and 29 g/d for C, I, PF, C_GH, I_GH, and PF_GH, respectively. Effects of GH were significant for fat accretion and plasma urea depression. Rectus femoris was highly refractory to catabolic effects of infection while psoas major was significantly catabolized during infection. Plasma concentrations of insulin-like growth factor-I (IGF-) I increased significantly in all GH-treated calves between Day 20 and 23 PI. Plasma IGF-I declined well below Day 20 values in all infected calves from the onset of the APR through the end of the study. The decrease in plasma IGF-I concentrations in I and I_G was highly correlated with the magnitude of the fever response. Hepatic mRNA for GH receptor and IGF-I was decreased in infected calves. Hepatic microsomal membrane binding of ¹²⁵I-GH did not differ between groups. The data suggest that effects of GH and parasitism on tissue metabolism during disease may vary among different specific tissue pools. The data demonstrate that daily GH administration in young calves does not prevent lean tissue losses and may accelerate fat depletion associated with cachectic parasitism. Furthermore, the onset of APR overrode the capacity for GH to maintain elevated plasma concentrations of IGF-I, an effect not readily explained through changes in GH-receptor binding.     

Effects of cimaterol administration on plasma concentrations of various hormones and metabolites in friesian steers

The aim of the experiment was to determine the acute and chronic effects of the β-agonist, cimaterol, on plasma hormone and metabolite concentrations in steers. Twelve Friesian steers (liveweight = 488 ± 3 kg) were randomly assigned to receive either 0 (control; n=6) or .09 mg cimaterol/kg body weight/day (treated; n=6). Steers were fed grass silage ad libitum. Cimaterol, dissolved in 140 ml of acidified distilled water (pH 4.2), was administered orally at 1400 hr each d. After 13 d of treatment with cimaterol or vehicle (days 1 to 13), all animals were treated with vehicle for a further 7 d (days 14 to 20). On days 1, 13 and 20, blood samples were collected at 20 min-intervals for 4 hr before and 8 hr after cimaterol or vehicle dosing. All samples were assayed for growth hormone (GH) and insulin, while samples taken at −4, −2, 0, +2, +4, +6 and +8 hr relative to dosing were assayed for thyroxine (T₄), triiodothyronine (T₃), cortisol, urea, glucose and non-esterified fatty acids (NEFA). Samples taken at −3 and +3 hr relative to dosing were assayed for IGF-I only. On day 1, cimaterol acutely reduced (P<.05) GH and urea concentrations (7.6 vs 2.9 ± 1.4 ng/ml; and 6.0 vs 4.9 ± 0.45 mmol/l, respectively; mean control vs mean treated ± pooled standard error of difference), and increased (P<.05) NEFA, glucose and insulin concentrations (160 vs 276 ± 22 μmol/l, 4.1 vs 6.2 ± 0.15 mmol/l and 29.9 vs 179.7 ± 13.9 μU/ml, respectively). Plasma IGF-I, T₃, T₄ and cortisol concentrations were not altered by treatment. On day 13, cimaterol increased (P<.05) GH and NEFA concentrations (7.7 vs 14.5 ± 1.4 ng/ml and 202 vs 310 ± 22 mEq/l, respectively) and reduced (P<.05) plasma IGF-I concentrations (1296 vs 776 ± 227 ng/ml). Seven-d withdrawal of cimaterol (day 20) returned hormone and metabolite concentrations to control values. It is concluded that : 1) cimaterol acutely increased insulin, glucose and NEFA and decreased GH and urea concentrations, 2) cimaterol chronically increased GH and NEFA and decreased IGF-I concentrations, and 3) there was no residual effect of cimaterol following a 7-d withdrawal period.     

Adenohypophyseal receptors for LHRH, pituitary content of gonadotropins and plasma concentrations of LH in cyclic, pregnant and postpartum beef cows

Adenohypophyseal concentrations of LHRH receptors, pituitary content of LH and FSH, and plasma concentrations of LH were determined in thirty Hereford, Angus or Hereford-Angus heifers that were randomly assigned by breed and weight to five periods including day 3 of the estrous cycle (CY), pregnant day 120 (P120), 200 (P200), 275 (P275), or day 2 postpartum (PP). Jugular blood samples were collected at 10-min intervals for 8 hr from all cows. Within 2 hr after completion of blood sampling, animals were slaughtered and the pituitary gland frozen at −196 C. LH pulse frequency/8 hr was reduced (P<.05) during gestation (.5, .2, and 1.5 ± .5/8 hr, for P120, P200, and P275, respectively) and PP (.5 ± .5/8 hr) compared to CY (7.8 ± .5/8 hr). Frequency of LH pulses/8 hr was not different (P>.1) among P120, P200 or PP periods but was different (P<.05) between P200 and P275. There were no differences in LH pulse height (P>.1) among periods; however, pulse amplitude was greatest (P<.05) at P120 (1.3 ± .2 ng/ml) and lowest between P200 and PP (.6 to .8 ± .2 ng/ml). Baseline concentrations of plasma LH did not differ (P>.1) among P and PP periods (.3 ± .1 ng/ml), but were lower (P<.05) than in CY animals (.7 ± .1 ng/ml). Concentration of adenohypophyseal LHRH receptors was approximately two-fold greater (P<.05) at P120 (25.85 ± 2.2 fmol/mg) than at all other periods (9.5 to 14.9 ± 2.2 fmol/mg). Pituitary content of LH was greatest at P120 (1.56 ± .11 ug/mg) and lowest (P<.05) at P275 and PP (0.46 to 0.52 ± .11 ug/mg). Pituitary content of FSH was greatest (P<.05) in P (12.7 to 17.0 ± 1.4 ug/mg) and PP (18.3 ± 1.4 ug/mg) vs CY (5.0 ± 1.4 ug/mg) cows and increased from P120 to PP (P<.05). Results indicate that physiological changes occurring during gestation may have an effect on subsequent function of the adenohypophysis in beef cows.     

Vaccination of cattle against bovine schistosomosis: current status and future prospects: a review

Bovine schistosomosis, caused by Schistosoma bovis, constitutes a serious veterinary problem in many parts of the world. The vaccination approaches for the control of bovine schistosomosis include the use of irradiation-attenuated S. bovis cercarial or schistosomular vaccines, S. bovis adult worms or whole-egg antigens and defined antigen vaccine. Irradiated S. bovis cercarial or schistosomular vaccines provide partial protection against S. bovis infection. However, this type of vaccine requires live infectious cercariae or viable schistosomula for induction of protection. Unfortunately, experimental immunizations with dead schistosome antigens have been largely unsuccessful. The surge of new techniques in cellular immunology and molecular biology has made possible the development of potential candidate vaccine antigens from various species of schistosomes including S. bovis. The efficiency of these vaccines has been evaluated in experimentally infected calves. These vaccines will probably replace the irradiated S. bovis vaccines. A broad-spectrum antischistosome vaccine which can kill a variety of human and animal schistosome species is yet to be produced.     

Diagnosis of Fasciola hepatica infection in beef calves by plasma enzyme analysis

Plasma analysis for albumin, total bilirubin, and total protein values and aspartate aminotransferase (AST), arginase, and gamma-glutamyl transpeptidase (GGT) activities was used for the early and quantitative diagnosis of experimental Fasciola hepatica infections in beef calves. Calves were infected on 3 occasions with 1,000 (n = 5), 100 (n = 5), or 10 (n = 4) metacercariae for a total infective dose of 3,000, 300, or 30, respectively. Albendazole (15 mg/kg of body weight) was administered to 7 infected calves on postinfection (initial) week (PIW) 13. All calves were euthanatized and necropsied on PIW 16 for the determination of fluke infections. Plasma constituents were determined weekly. Significant (P less than 0.05) increases in AST activity occurred as early as PIW 4 and GGT activity at PIW 9, as compared with that in noninfected controls. Fluke burden-related differences were observed in GGT activity from PIW 9 onward. Increases in AST activity reflected parenchymal liver damage, whereas increases in GGT reflected hepatobiliary damage; therefore, differentiation could be made between the migratory and ductal phases of the infection. There was no correlation between arginase activity and fluke infection. As compared with fecal examination results, plasma enzyme analysis gave an earlier and semiquantitative indication of F hepatica infection in experimentally infected calves. Although increases in these plasma constituents were not definitely diagnostic of fascioliasis, useful information on the size of the fluke burden and progress of the disease process could be obtained by these methods. Plasma enzyme analyses of AST and GGT were not indicative of chemotherapeutic success or failure when calves with mature F hepatica (14 weeks old) infections were treated.     

PGD2/DP1途径对细菌感染奶牛子宫内膜组织中HMGB-1和PAFR表达的影响

为了研究前列腺素D₂（prostaglandin D₂,PGD₂）/DP₁受体途径对大肠杆菌（Escherichia coli）和金黄色葡萄球菌（Staphylococcus aureus）感染奶牛子宫内膜组织中炎症介质HMGB-1和PAFR的表达及对组织损伤程度的影响,试验以体外培养奶牛子宫内膜组织为研究对象,应用1×10^-6 mol/L DP₁受体激动剂（BW-245C和15d-PGJ2）和等量（1×10⁶ CFU/mL）大肠杆菌、金黄色葡萄球菌共同处理奶牛子宫内膜组织,采用实时荧光定量PCR、免疫组化和HE染色法检测奶牛子宫内膜组织中HMGB-1和PAFR的表达并评价组织损伤情况。结果显示,与空白对照组相比,大肠杆菌和金黄色葡萄球菌感染奶牛子宫内膜组织中HMGB-1和PAFR表达量显著升高（P<0.05）,而DP₁受体激动剂与大肠杆菌和金黄色葡萄球菌共处理奶牛子宫内膜组织中DP₁受体激动剂显著抑制奶牛子宫组织中HMGB-1和PAFR的表达（P<0.05）。HE染色结果显示,大肠杆菌与金黄色葡萄球菌感染的奶牛子宫内膜组织上皮细胞和腺上皮细胞完全脱落、坏死、崩解;而DP₁受体激动剂、大肠杆菌、金黄色葡萄球菌共同处理奶牛子宫内膜组织中,DP₁受体激动剂的加入显著减轻奶牛子宫内膜组织的损伤程度（P<0.05）。免疫组化染色法结果与以上两种方法结果一致。结果表明,PGD₂能够抑制大肠杆菌与金黄色葡萄球菌感染的奶牛子宫内膜组织中损伤相关因子HMGB-1、PAFR的表达,减轻组织损伤程度,这一作用可能是由DP₁受体所介导的。     

The effect of immunization against somatostatin on growth and concentration of somatotropin in plasma of Holstein calves

Holstein calves were used to investigate the effects of immunization procedures against somatostatin (SRIF) on growth and concentrations of somatotropin in plasma. In Trial 1, eight heifers 37 weeks of age were inoculated with cyclic-SRIF conjugated to human alpha-globulin. Final body weight, average daily gain, and measurements of body size were not significantly different between control and SRIF-immunized calves. Apparent total tract nutrient digestibilities and efficiency of feed utilization also were not significantly different between treatments. Plasma concentrations of somatotropin were increased and plasma concentrations of urea nitrogen were decreased in calves immunized against SRIF compared to controls, but these mean differences were not significant. In Trial 2, eleven bull calves seven weeks of age were inoculated with cyclic-SRIF conjugated to keyhole limpet hemocyanin. Calves immunized against SRIF had larger average daily gains (P less than .06) than did control calves. Body size, efficiency of feed utilization, and concentrations of somatotropin in plasma were not significantly different for SRIF immunized calves and control calves. Urea nitrogen in plasma was lower (P less than .04) for calves immunized against somatostatin than for control calves. Data indicate that Holstein calves can produce auto-antibodies against SRIF; however, additional research will be required before such immunization techniques can be effectively used to improve weight gains in cattle.     

Pancreatic insulin secretory response and insulin action in heat-exposed sheep given a concentrate or roughage diet

The effects of heat exposure and type of diet on the insulin secretory response to glucose and glucose disposal in response to insulin action in female sheep were investigated employing hyperglycemic and euglycemic clamp techniques. Animals were divided into concentrate and roughage diet groups, and were maintained at the same intake levels of metabolizable energy and crude protein in both diets. Each diet group was subjected to either thermoneutral (20°C, 70% RH) or hot (30°C, 70% RH) environment, followed by glucose clamp experiments.

Heat-exposed sheep showed significant increases in respiration rates (P<.001) and rectal temperature (P<.05). Plasma glucose concentrations in the basal conditions were lower (P<.01) in the hot environment than in the thermoneutral environment, but there was no significant difference in basal levels of plasma insulin between the environmental treatments. In the hyperglycemic clamp experiment, mean plasma insulin increments increased (P<.05) during the heat exposure period across diet treatments. The ratio of mean plasma insulin increment to glucose infusion rate tended to be higher (P<.07) in the hot environment than in the thermoneutral environment, but diet treatment did not affect the ratio of mean plasma insulin increment to glucose infusion rate. The euglycemic clamp technique showed that glucose infusion rates remained unchanged among treatments. Insulin secretion response to glucose could be stimulated in the hot environment.     

Performance and nitrogen metabolism of calves fed conventionally or following an enhanced-growth feeding program during the preweaning period

Thirty-seven Holstein and seven crossbred female calves (16.1 ± 4.60 days, and an initial BW of 36.5 ± 3.19) were used to study the effects of conventional (CF) vs enhanced-growth feeding programs (EF) on performance, plasma amino acid (AA) concentrations, and rumen microbial development. After 1 week of adaptation to milk replacer (MR), the CF calves received 4 l/day of MR at 12.5% DM throughout the preweaning period, and the EF calves were offered MR at 18% DM: 6 l/day from 1 to 6 days, 8 l/day from 7 to 26 days, and 4 l/day from 27 days to weaning day (38 days). Calf starter and water were offered ad libitum throughout the study (87 days). Calves fed EF were heavier (P < 0.05) than CF calves at the end of the study (111.7 vs 102.6 ± 1.72 kg, respectively). Until the 27 days, average daily gain (ADG) was greater (P < 0.001) for EF than for CF calves (1.00 vs 0.49 ± 0.061 kg/day, respectively), but it was lower (P < 0.001) from days 27 to 45 of the study (0.32 vs 0.71 ± 0.061 kg/day, respectively), coinciding with the days around weaning. Starter intake was greater (P < 0.001) for CF than for EF calves during the first 45 days of the study (0.60 vs 0.27 ± 0.061 kg/day, respectively) but similar afterwards. As a consequence, EF treatment may have delayed rumen function as suggested by total daily purine derivatives urinary excretion (49.52 vs 33.27 ± 3.095 mmol/day, in CF and EF calves, respectively). Linear regression analyses showed a positive relationship between plasma Trp and Phe concentrations and ADG, and a negative relationship between these two AA and plasma urea concentrations, suggesting that Trp and Phe could be limiting growth in calves fed conventional feeding programs.     

Alterations of serum insulin-like growth factor-I (IGF-I) and IGF-binding proteins (IGFBPs) in swine infected with the protozoan parasite Sarcocystis miescheriana.

The effects of a Sarcocystis miescheriana infection on insulin-like growth factor-I (IGF-I) and insulin-like growth factor binding proteins (IGFBPs) were investigated to determine possible mechanisms of growth retardation in growing pigs. Sixteen pigs averaging 14 kg body weight were divided into 4 groups of 4 pigs each and infected either with 0.5, 1.0, or 3.0 × 10⁶ sporocysts of S. miescheriana. Four pigs were retained as non-infected controls; however, they became serologically positive during the course of the infection. Effects also were investigated in 2 groups of 3 pregnant sows. One group was infected with 0.5 × 10⁶ sporocysts and the other group was retained as uninfected controls. Body weights of infected growing pigs were depressed as compared to controls following the acute phase 15 d after infection (dai). Serum concentrations of IGF-I dropped significantly (p < 0.05) during the acute phase of infection in all infected groups of growing pigs. Conversely, the amounts of unsaturated serum IGFBPs were elevated significantly (p < 0.05) during the acute phase of infection. Specifically, serum concentrations of IGFBP-1, IGFBP-2, and IGFBP-4 were elevated at this time, as determined by ligand blot analysis. There was no association between growth factor alterations and tissue damage as measured by serum creatinine kinase and aspartate aminotransferase levels. The extent of effects in growing pigs was related to the amount of the original parasite inoculum.

During the acute phase of infection 2 of 3 pregnant sows aborted. The third sow went to term, but piglets were stillborn or died within 24 hr. Compared to uninfected controls, serum concentrations of IGF-I in infected pregnant sows were depressed during and after the acute phase of the infection. Levels of unsaturated serum IGFBPs in pregnant sows were not affected.

These data suggest that decreased IGF-I levels and/or elevated levels of specific forms of IGFBPs may be a mechanism by which growth is affected in feeder pigs infected with S. miescheriana.     

Neuroendocrine regulation of growth hormone secretion in sheep. IV. Central and peripheral cholecystokinin

The result of alterations in the levels of CCK, in the blood and in the cerebrospinal fluid, on the functioning of the growth hormone axis has been examined in sheep. Male Coopworth sheep of about 40 kg liveweight were given various doses of CCK either intracerebroventricularly (icv) or intravenously (iv). Other similar sheep were given various doses of a CCK antagonist (loxiglumide) by the same routes. Bolus iv administration of either 35 μg or 200 μg of CCK had no effect on plasma GH levels. When given icv, however, CCK resulted in a marked (P<0.01) prolonged depression in plasma GH levels. The decrease in GH secretion could be partially attenuated by concurrent administration of loxiglumide, but was completely unaffected by concurrent administration of anti-somatostatin serum icv. Loxiglumide alone had no effect on plasma GH levels when given at up to 200 μg icv, but intravenous administration of 8 mg of the CCK antagonist resulted in an increase in plasma GH concentrations (P<0.05). Plasma levels of somatostatin, glucose and cortisol were unaffected by both icv and iv administration of CCK. These results show that CCK can have a strong GH-inhibiting effect in the brain. Furthermore, this effect seems to be independent of hypothalamic somatostatin, suggesting another GH-inhibiting system exists.     

Serum prolactin response to thyrotropin releasing hormone in estrogen treated hypophysial stalk-transected gilts

Twelve crossbred gilts, 169 ± 3 days of age and 72.8 ± 3.4 kg body weight, were hypophysial stalk-transected (HST)1 or sham hypophysial stalk-transected (S-HST). Gilts were ovariectomized 6 days later and assigned to four treatments of 3 gilts each in a 2 × 2 factorial arrangement. One-half of the HST and S-HST gilts received 5 mg estradiolbenzoate (EB) or corn oil vehicle im at 0800 hr daily for 5 days beginning 64 ± 3 days after HST or S-HST. Blood was collected by jugular vein cannula at 0830 and 0900 hr the day after the last injection of EB or oil. Immediately after the 0900 hr sample, 200 μg thyrotropin releasing hormone (TRH) were injected (iv). Mean basal serum prolactin (PRL) concentration was similar for HST (10.3 ± 1.0 ng/ml) and S-HST (12.3 ± 1.7 ng/ml) gilts, however mean basal serum PRL concentration was greater (P<.05) for EB-treated gilts (13.7 ± 1.3 ng/ml) than for oil-treated gilts (8.8 ± .5 ng/ml). Mean serum PRL concentration of all gilts increased within 10 min and returned to approximately 20 ng/ml by 150 min after TRH. Maximum serum PRL concentrations at 10 min after TRH were greater (P<.01) for S-HST (255.9 ± 29.6 ng/ml) than HST gilts (83.4 ± 18.8 ng/ml), but were not different for EB (198.0 ± 50.6 ng/ml) and oil-treated gilts (141.4 ± 36.3 ng/ml). Area under the serum PRL response curve after TRH was greater (P<.005) for S-HST than HST gilts and for EB than oil-treated gilts (P<.05). These results do not eliminate the possible influence of estrogen on PRL secretion at the hypothalamus, but do indicate that estrogen directly stimulated the anterior pituitary gland to secrete PRL.     

金黄色葡萄球菌对BV2细胞IFN-α生成的影响

旨在探究金黄色葡萄球菌（Staphylococcus aureus，简称金葡菌）对小胶质细胞α干扰素（interferon α，IFN-α）生成的影响，本研究先用金葡菌感染BV2细胞，检测IFN-α的mRNA表达、分泌到细胞培养液上清的量以及TBK1/IRF3通路的激活情况，再分别用TBK1抑制剂BX-795和amlexanox、NF-κB抑制剂IMD-0354处理细胞，检测TBK1/IRF3的激活以及IFN-α水平。结果表明，BV2感染金葡菌后，IFN-α转录水平在3~12 h升高，6~12 h释放量增加，呈剂量依赖性，TBK1和IRF3的转录水平和蛋白水平未发生变化，但在1~12 h其磷酸化水平升高，表明金葡菌激活BV2细胞TBK1/IRF3通路，BX-795、amlexanox和IMD-0354处理细胞后，IRF3磷酸化水平降低，IFN-α生成减少，表明TBK1和NF-κB参与金葡菌促进BV2细胞生成IFN-α。综上所述，金葡菌感染BV2后促进IFN-α生成，且该过程依赖于TBK1和NF-κB，本结果为临床防治中枢神经系统感染金葡菌提供了可能靶点和理论基础。     

Changes in food intake and circulating leptin due to gastrointestinal parasitism in lambs of two breeds

A reduction in food intake is a prominent feature of many infectious diseases. However, the underlying mechanisms of parasite-induced anorexia in sheep are poorly understood. Here, we tested the hypotheses (a) that the degree of parasite-induced anorexia in lambs is influenced by their growth potential and (b) that nematode infection results in elevated plasma leptin concentration in lambs. The hypotheses were tested with Suffolk x Greyface (S) and Scottish Black-face (B) lambs that are known to differ in their growth potential (S lambs are of greater growth potential than B lambs). During a primary parasite infection, 24 out of 48 lambs per breed were trickle-infected with 7,000 infective Teladorsagia circumcincta larvae per day, 3 d/wk, for a period of 12 wk (experiment I). The lambs were then dewormed, and after a 2-wk interval, half of the 24 lambs per breed that were previously infected were reinfected for another 12 wk with the same parasite and dose as used in the primary infection (experiment II). In both experiments, infected lambs were fed grass pellets for ad libitum intake, whereas noninfected lambs were fed grass pellets for either ad libitum or restricted intakes. The S lambs were more susceptible than B lambs to nematode infection, as judged from the differences in fecal egg counts (P = 0.007). Parasitized lambs of the more susceptible breed (S) showed anorexia [i.e., a decrease in intake of 13% compared with uninfected controls (P = 0.01)], whereas no significant reduction in food intake was observed in lambs of the more resistant breed (B). Reexposure to nematode infection of previously infected animals tended to result in renewed anorexia in S lambs but not in B lambs (P = 0.08) in a similar extent as during primary infection. Plasma leptin concentrations did not differ between ad libitum-fed infected and control lambs but were greater in infected than in noninfected lambs at a similar level of food intake during both the primary (P = 0.02) and the secondary parasitic infection (P = 0.004) in both breeds. The results show that leptin may be involved in the response of lambs to infection but that it is unlikely that leptin alone is responsible for the parasite-induced anorexia in lambs.     

Evaluation of Therapeutic Effect of Porcine β-defensin-2 on Streptococcus suis Infection

Defensin is a small cationic peptide widely distributed in animals and plants. It is an important component of the innate immune defense system of mammals. It has a broad spectrum of antibacterial, antiviral and antifungal activities. Porcine β-defensin-2 (PBD-2) is one of the natural defensins expressed in pigs and has good antibacterial activity in vitro. The purpose of this study is to evaluate the therapeutic effect of PBD-2 in the treatment of Streptococcus suis infection in animals and to provide more insights for evaluating the value of PBD-2 as a therapeutic drug. Firstly, the bactericidal activity of PBD-2 against S. suis clinical isolate SC-19 was measured in vitro, and the cytotoxicity of PBD-2 was tested on mouse-derived macrophages (RAW264.7). Subsequently, C57 female mice aged 4-6 weeks and weighing between 18-22 g infected with S. suis SC-19 were treated with PBD-2 or PBS (n ≥ 6). The survival rate of mice, and the bacteria loads, inflammatory cytokine levels, and pathological changes of tissues were determined. The results showed that PBD-2 (25-200 μg·mL^-1) could significantly inhibit the growth of S. suis SC-19 in a concentration-dependent manner (P<0.05). At the same time, PBD-2 had no significant toxic effect on RAW cells (P>0.05). The in vivo study revealed that PBD-2 treatment could effectively reduce the mortality of mice infected with S. suis SC-19. At the same time, PBD-2 treatment significantly reduced bacteria loads in brain, lung, spleen and blood of mice, and decreased the levels of inflammatory cytokines IL-6, IL-12 and TNF-α in mouse sera (P<0.05). At the same time, PBD-2 treatment also significantly alleviated the degree of pathological damages in lung and spleen tissues of mice infected with bacteria (P<0.05). These results indicate that PBD-2 confers great resilience to S. suis in vitro without significant cytotoxicity, while it also exhibits a therapeutic effect on S. suis infection in mice, suggesting that the use of PBD-2 as a therapeutic drug and substitutes for antibiotics is of an excellent prospect.     

金黄色葡萄球菌处理对奶牛乳腺上皮细胞外泌体表征的影响

【目的】本研究旨在从金黄色葡萄球菌(Staphylococcus aureus,S.aureus,以下简称金葡菌)处理奶牛乳腺上皮细胞(bovine mammary epithelial cell,BMEC)上清中分离外泌体,探究金葡菌与BMEC数量比值(MOI)和处理后培养时间对外泌体总浓度的影响,并对金葡菌诱导BMEC释放的外泌体进行分离和鉴定。【方法】采用金葡菌临床分离菌株以MOI＝1和MOI＝10分别处理BMEC 3 h,对已处理的BMEC继续无外泌体培养9、12、24 h,同时设空白对照组,通过扫描电镜观察金葡菌处理对BMEC造成的超微结构损伤。采用超速离心法分离金葡菌诱导的BMEC上清液中的外泌体,分别用透射电镜、纳米颗粒追踪分析技术及Western blotting法对外泌体形态、颗粒大小和特异性标志蛋白进行分析鉴定。通过检测外泌体总蛋白浓度评估不同MOI和处理后培养时间对外泌体总量的影响,明确金葡菌诱导BMEC释放外泌体的最佳条件。【结果】通过扫描电镜观察发现,金葡菌处理BMEC后,细胞微绒毛脱落,细胞骨架破坏。透射电镜观察分离的外泌体为圆形的双层膜囊泡,直径在30~150 nm,形态均一。通过检测外泌体总蛋白浓度显示,在MOI＝10时,培养9、12和24 h金葡菌诱导BMEC释放外泌体蛋白浓度均高于MOI＝1,而培养12 h时释放外泌体蛋白浓度最高。对金葡菌诱导BMEC释放的外泌体进一步鉴定,其平均直径约为116 nm;Western blotting检测结果显示外泌体标记物CD9、CD81和TSG101表达阳性。【结论】本研究从形态学和分子生物学特征等方面证实获得的分离物为外泌体;金葡菌可以诱导BMEC释放外泌体,当MOI＝10时,金葡菌处理细胞3 h后继续无外泌体培养12 h,收获的细胞上清中外泌体蛋白含量最高。     

Distribution of ceftiofur into Mannheimia haemolytica‐infected tissue chambers and lung after subcutaneous administration of ceftiofur crystalline free acid sterile suspension

Washburn, K., Johnson, R., Clarke, C, Anderson, K. Distribution of ceftiofur into Mannheimia haemolytica‐infected tissue chambers and lung after subcutaneous administration of ceftiofur crystalline free acid sterile suspension. J. vet. Pharmacol. Therap. 33 , 141–146. The objective of this study was to evaluate the penetration of ceftiofur‐ and desfuroylceftiofur‐related metabolites (DCA) into sterile and infected tissue chambers, lung tissue and disposition of DCA in plasma across four different sacrifice days postdosing. Twelve healthy calves were utilized following implantation with tissue chambers in the paralumbar fossa. Tissue chambers in each calf were randomly inoculated with either Mannheimia haemolytica or sterile PBS. All calves were dosed with ceftiofur crystalline free acid sterile suspension (CCFA‐SS) subcutaneously in the ear pinna. Calves were randomly assigned to 4 groups of 3 to be sacrificed on days 3, 5, 7 and 9 postdosing. Prior to euthanasia, plasma and tissue chamber fluid were collected, and immediately following euthanasia, lung tissue samples were obtained from four different anatomical sites DCA concentration analysis. Results of our study found that, in general, DCA concentrations followed a rank order of plasma > infected tissue chamber fluid > noninfected tissue chamber fluid > lung tissue. Data also indicated DCA concentrations remained above the therapeutic threshold of 0.2 μg/mL for plasma and chamber fluid and 0.2 μg/g for lung tissue for at least 7 days post‐treatment.