Plasma levels of cortisol and corticosteroid-binding globulin (CBG) and hepatic CBG mRNA expression in pre- and postnatal pigs

The relationships among hepatic corticosteroid-binding globulin (CBG) mRNA expression and plasma concentrations of cortisol and CBG was evaluated in fetal pigs (n=7-14 per age) on days 50, 70, 80, 90, and 104 of gestation and postnatal pigs (n=8 per age) on days 1, 3, 10, 20, 30, and 40 following birth. In fetal pigs, hepatic CBG mRNA expression was highest (P<0.01) on day 50 as compared to days 90 and 104, exhibiting an overall negative relationship (r=-0.63; P<0.01) with estimated gestation age. Plasma porcine CBG (pCBG) concentration was correlated (r=0.34; P<0.05) with hepatic CBG mRNA level. Plasma cortisol concentrations were not different over this same period. In postnatal pigs, hepatic CBG mRNA expression increased (P<0.01) from days 3 to 40. The pCBG concentration increased (P<0.01) from days 1 (6.1+/-3.4 microg/ml) to 10 (15.1+/-3.7 microg/ml), while plasma cortisol concentration remained constant. An understanding of the relation between hepatic CBG mRNA and circulating pCBG concentrations may provide insight into the mechanisms determining the bioavailability of cortisol necessary in prenatal development and the conservation of cortisol during postnatal development in the pig.     

Effect of menhaden fish oil supplementation and lipopolysaccharide exposure on nursery pigs. I. Effects on the immune axis when fed diets containing spray-dried plasma

The objective of the present study was to evaluate the potential immunological benefit of adding menhaden fish oil to the diet of weaned pigs. Twenty-four crossbred male pigs were weaned at approximately 18 days of age and placed on a complex nursery diet containing 30% lactose and 7% plasma protein with 6% corn oil as the fat source (Cont, n=12) or with 5% menhaden fish oil and 1% corn oil as the fat source (MFO, n=12) for a period of 15 days. Body weights did not differ (P>0.78) between dietary groups either at the beginning or end of the 15 days feeding period. On day 15, all pigs were non-surgically fitted with an indwelling jugular catheter. On d 16, pigs received an i.v. injection of either saline (n=6/dietary group) or lipopolysaccharide (LPS; 150 μg/kg body weight; n=6/dietary group) and blood samples were collected at 30 min intervals for a period of 5 h. Serum was harvested and stored at −80 °C for analysis of cortisol (CS), corticosteroid-binding globulin (CBG), tumor necrosis factor-alpha (TNF-) and interferon-gamma (IFN-γ). There was no significant effect of diet on basal concentrations (Time 0) of any of the blood parameters analyzed. A Time×Treatment×Diet interaction (P<0.03) was observed for serum CS such that those pigs which consumed the MFO diet followed by LPS treatment had a reduced CS response as compared to the LPS-treated pigs on the Cont diet. A Time×Treatment interaction (P<0.01) was observed for serum CBG such that LPS treatment reduced circulating CBG as compared to the saline-treated pigs. Time×Treatment×Diet interactions were also observed for serum concentrations of TNF- (P=0.084) and IFN-γ (P=0.022) such that both the TNF- and IFN-γ response to the LPS challenge was lower in those pigs receiving the MFO diet as compared to the LPS-treated pigs on the Cont diet. Overall, serum CS was negatively correlated with the CBG response (r=−0.40, P<0.001), however, the strongest negative correlation was observed in the LPS-treated pigs which consumed the MFO diet (r=−0.63, P<0.001). While further studies are needed to evaluate the immunological response of including MFO in the nursery pig diet, the present study demonstrates that supplementation with MFO does indeed alter the immunological response to an LPS challenge.     

Plasma concentrations of ACTH precursors correlate with pituitary size and resistance to dexamethasone in dogs with pituitary-dependent hyperadrenocorticism

This study was performed to determine whether in dogs with pituitary-dependent hyperadrenocorticism (PDH) excessive release of adrenocorticotrophic hormone (ACTH) is accompanied by secretion of ACTH precursor molecules. In addition, we investigated whether the plasma ACTH precursor concentrations were correlated with the size of the pituitary gland and with the degree of resistance to negative glucocorticoid feedback. In 72 dogs with PDH, the plasma ACTH precursor concentration was determined by calculating the difference between the results of a radioimmunoassay (RIA) in which besides ACTH, ACTH precursors were also measured and a highly specific immunoradiometric assay (IRMA) using a polyclonal antibody against ACTH. The degree of resistance to glucocorticoid feedback was established by determining the effect of dexamethasone administration (0.1 mg/kg) on the urinary corticoid/creatinine ratio. The pituitary height/brain area (P/B) ratio, determined by computed tomography, was used as a measure for the size of the pituitary gland. The plasma ACTH precursors concentration ranged from 18 to 2233 ng/L (median 93 ng/L). In 38 dogs, the pituitary was enlarged and plasma ACTH precursors concentrations in these dogs (median 130 ng/L, range 24–2233 ng/L) were significantly (P<0.05) higher than those in the dogs without pituitary enlargement (median 72 ng/L, range 18–481 ng/L). In concordance, P/B ratios correlated significantly with plasma ACTH precursor concentrations (r=0.35, P<0.01). In addition, the P/B ratios were significantly correlated with the degree of dexamethasone resistance (r=0.42, P<0.001). Plasma ACTH precursor concentrations in the dexamethasone-resistant dogs (median 210 ng/L, range 24–628 ng/L) were significantly higher (P<0.01) than those in the dexamethasone-sensitive dogs (median 72 ng/L, range 18–2233 ng/L). Similarly, the degree of dexamethasone resistance was also significantly correlated with the plasma ACTH precursor concentrations (r=0.33, P<0.01). Dogs with an elevated plasma -MSH concentration (n=14) had significantly (P<0.001) higher plasma ACTH precursor concentrations (median 271 ng/L, range 86–2233 ng/L) than dogs with non-elevated -MSH (median 73 ng/L, range 18–481 ng/L). In addition, the plasma concentrations of -MSH correlated significantly with both plasma ACTH precursor concentrations (r=0.53, P<0.001) and P/B ratios (r=0.26, P<0.05). In conclusion, in all dogs with PDH the ACTH concentrations determined by the RIA were higher than the concentrations measured by IRMA indicating the presence of circulating ACTH precursors. High plasma ACTH precursor concentrations were especially found in dexamethasone-resistant dogs with large corticotroph adenomas, some of them probably of PI origin. In the association of large corticotroph adenoma, dexamethasone resistance and high plasma concentrations of ACTH precursors, the decreased sensitivity of the corticotroph cells to glucocorticoid feedback may play a pivotal role.     

Hepatic corticosteroid-binding globulin (CBG) messenger RNA expression and plasma CBG concentrations in young pigs in response to heat and social stress

Plasma cortisol, porcine corticosteroid-binding globulin (pCBG), hepatic CBG expression, and other physiological and behavioral measures of stress were studied in pigs in response to elevated temperature in conjunction with establishing a social hierarchy. Twenty-four crossbred pigs were weaned at 25 d of age (three or six pigs from six sows) and housed in littermate groups at 23 +/- 2 degrees C. At 57 d of age (d 0), animals were weighed and placed under general anesthesia for collection of blood (10 mL) and liver (approximately 100 mg) samples. On d 1, three unacquainted pigs of similar BW (23 +/- 1 kg) from different litters were allotted to each of eight nursery pens within two environmentally controlled rooms (12 pigs per room). From d 1 to 7, one room was maintained at 23 +/- 2 degrees C (CON) and the other at 33 +/- 2 degrees C (HEAT). Both rooms were kept at 23 +/- 2 degrees C from d 8 to 14. Animals were videotaped for 72 h beginning on d 1 and 8 to document behavioral changes in response to room temperature. The social hierarchy of pigs within each pen was based on fight activity recorded on d 1 to 3. Blood and liver tissue were collected again on d 7 and 14. The ADG for HEAT pigs increased (P < 0.05) over d 8 to 14 compared with d 1 to 7. In contrast to CON pigs, HEAT pigs displayed increased (P < 0.01) drinking but decreased feeding and lying in contact with other pigs from d 1 to 3, and similar drinking and feeding but increased (P < 0.01) lying with contact behaviors from d 8 to 10. With the exception of subordinate pigs exhibiting less (P < 0.05) frequent standing/walking behavior than the dominant or intermediate pigs on d 1 to 3, frequency of behaviors for both recorded time periods did not differ among pigs due to social status, regardless of treatment. The concentration of plasma haptoglobin in HEAT pigs on d 7 compared with d 0 increased (467 vs. 763 mg/L; P < 0.05), whereas cortisol and pCBG decreased (274 vs. 235 nmol/L and 11.4 vs. 9.9 mg/L, respectively; P < 0.05) as a result of treatment. The free cortisol index (total cortisol/pCBG) was greater (P < 0.05) in HEAT pigs on d 14 than on d 0 or 7. Hepatic CBG mRNA level was not affected by treatment. On d 14, HEAT pigs had plasma cortisol, pCBG, and haptoglobin concentrations similar to those of CON pigs. These results indicate that measured behavioral and physiological responses were not related to social status, and decreased circulating levels of cortisol and pCBG in pigs following a 7-d exposure to elevated temperature may not be determined by hepatic CBG mRNA expression.     

The use of purine derivatives/creatinine ratio in spot urine samples as an index of microbial protein supply in Yerli Kara crossbred cattle

Three experiments were conducted to evaluate the use of purine derivatives (PD)/creatinine ratio in spot urine samples as an index of microbial protein supply in Yerli Kara crossbred cattle (YK-C). In Experiment I, response of daily PD excretion to feed intake in YK-C at state farm was measured. In Experiment II, spot urine sampling techniques was applied at state farm and four YK-C bulls were used. In Experiment III, spot urine sampling technique was applied at small-holder farms. There were significant correlations (R² = 0.99) between PD excretion (mmol/day) and digestible organic matter intake (DOMI) (kg/day) in Experiment I, Y = 12.5 (± 0.5) + 19.7 (± 3.5)X (R² = 0.99, n = 16). The equation obtained from Experiment I could be expressed as: Y = − 2.3 (± 0.3) + 0.953 (± 0.06)X, (R² = 0.99, n = 49) where Y is PD excretion (mmol/day) and X is the PDC index. The PDC index was calculated as the molar concentration ratio of PD to creatinine times the metabolic body weight (kg). The corresponding microbial-N values to PDC index of groups I, II and III in developed banding system are 15–25 g/day. Experimentally estimated DOMI was 2.21 ± 0.15 kg/day. Estimated DOMI of groups I, II, and III were 2.8 ± 0.6, 2.6 ± 0.7 and 2.7 ± 0.7 kg/day, respectively. In conclusion, the PDC index in spot urine samples could be used under similar farm condition as an indicator of microbial protein supply in YK-C cattle. Estimated DOMI from PDC index in spot urine samples under defined field conditions may help the development of feeding strategies for YK-C cattle held by small holders.     

Duration of immunity of a quadrivalent vaccine against respiratory diseases caused by BHV-1, PI3V, BVDV, and BRSV in experimentally infected calves

Several laboratory studies assessed the duration of immunity of a quadrivalent vaccine (Rispoval™4, Pfizer Animal Health) against bovine respiratory diseases (BRD) caused by bovine herpes-virus type-1 (BHV-1), parainfluenza type-3 virus (PI₃V), bovine viral-diarrhoea virus type 1 (BVDV), or bovine respiratory syncytial virus (BRSV). Calves between 7 weeks and 6 months of age were allocated to treatment and then were injected with two doses of either the vaccine or the placebo 3 weeks apart. Six to 12 months after the second injection, animals were challenged with BHV-1 (n = 16), PI₃V (n = 31), BVDV (n = 16), or BRSV (n = 20) and the course of viral infection was monitored by serological, haematological (in the BVDV study only), clinical, and virological means for ≥2 weeks. Infection induced mild clinical signs of respiratory disease and elevated rectal temperature in both vaccinated and control animals and was followed by a dramatic rise in neutralising antibodies in all treatment groups. Titres reached higher levels in vaccinated calves than in control calves after challenge with BHV-1, BVDV, or BRSV. On day 3 after PI₃V challenge, virus shedding was reduced from 3.64 log₁₀ TCID₅₀ in control animals to 2.59 log₁₀ TCID₅₀ in vaccinated animals. On days 6 and 8 after BRSV challenge, there were fewer vaccinated animals (n = 2/10 and 0/10, respectively) shedding the virus than control animals (n = 8/10 and 3/10, respectively). Moreover, after challenge, the mean duration of virus shedding was reduced from 3.8 days in control animals to 1 day in vaccinated animals in the BVDV study and from 3.4 days in control animals to 1.2 days in vaccinated animals in the BRSV study. The duration of immunity of ≥6 months for PI₃V, BHV-1 and BVDV, and 12 months for BRSV, after vaccination with Rispoval™4, was associated mainly with enhanced post-challenge antibody response to all four viruses and reduction of the amount or duration of virus shedding or both.     

Intermittent suckling improves post-weaning feed uptake but does not change functional gut characteristics of piglets

Suckling pigs were separated from their dam for 24 h on day 21 (1 × 24 h fasting, n = 10) or day 21 and 24 (2 × 24 h fasting, n = 10). Pigs in the control group (n = 10) were not fasted before weaning. All pigs were weaned on day 28 postpartum. Feed intake during the first 4 days post-weaning was higher (P < 0.05) for pigs exposed to 1 × 24 h fasting compared with controls. Water consumption was not affected by fasting prior to weaning. The difference in post-weaning feed uptake was not reflected in any clinical traits, intestinal morphology, or activity of digestive enzymes (maltase, dipeptidylpeptidase IV, aminopeptidases A and N; P > 0.15). In conclusion, a short period of fasting prior to weaning can increase post-weaning feed uptake, although this had no clinical impact under the present experimental conditions.     

Pathophysiological and immune cell responses in calves prior to and following lung challenge with formalin-killed Pasteurella multocida biotype A:3 and protection studies involving subsequent homologous live challenge

Pneumonic pasteurellosis is a common respiratory infection in cattle that has major economic and welfare implications world-wide and the incidence in the UK due to Pasteurella multocida, currently the same as that associated with Mannheimia haemolytica, is increasing. Whereas much is known regarding the pathogenesis of M. haemolytica infections little information is available on the pathogenic process of pasteurellosis initiated by P. multocida. In the present work calf systemic and innate immune responses to intratracheal challenge with formalin-killed P. multocida biotype A:3 and to subsequent experimental lung infection with live P. multocida were investigated. Eight-week-old calves were challenged intratracheally on day 0 with either 10⁹ colony forming units (cfu) of formalin-killed P. multocida biotype A:3 in 300 ml saline (n=10) or 300 ml saline alone (n=10), followed, at day 21, by challenge with 10⁹ cfu live P. multocida. Pathophysiological and lung phagocyte responses were assessed by clinical monitoring, sequential lung lavage and blood sampling. Results for samples obtained before, during and after challenge showed clinical and acute phase protein responses to both bacterial culture and saline control treatments, although higher responses were associated with bacterial challenge. Phagocytosis of P. multocida during 1 h incubation periods with lavaged cells in vitro was unaffected by exposure in vivo to killed P. multocida and there was evidence that P. multocida was able to survive intracellularly during this assay. There was no indication that lung exposure to formalin-killed P. multocida conferred protection against subsequent homologous live challenge.     

Effect of constant-release melatonin implants and prolonged exposure to a long day photoperiod on prolactin secretion and hair growth in mouflon (Ovis gmelini musimon)

The aims of this study were to examine whether mouflons exposed to constant long and short day photoperiods are able to exhibit an annual cycle of hair growth and moult, and prolactin (PRL) secretion. Mouflon ewes were assigned to three groups of treatment. Ewes were maintained, either under natural photoperiod (control, n=9), or received a series of subcutaneous melatonin implants from December to April (n=8), or were exposed to a constant long day photoperiod (16-h light:8-h dark; 16L:8D) during 18 months (n=7). Blood was collected weekly to determine PRL concentrations, and hair samples were clipped weekly from the base of the neck to measure the length of predominant hair. Under constant long days and with melatonin implants, mouflons expressed an annual rhythm of PRL secretion, even though these treatments modified the times of rise or falling of PRL concentrations throughout the year. Hair growth initiation was almost coincident with the summer solstice in both control and melatonin-implanted mouflons but occurred two months earlier in long day hold mouflons (P<0.001). Long day hold mouflons had a lower hair growth rate than control and melatonin-implanted mouflons (P<0.001), and at the end of the experiment, a shorter hair length (3.4±0.24 cm; P<0.01) than control (4.3±0.17 cm), and melatonin-implanted mouflons (4.2±0.12 cm). Our data support the conclusion that in mouflon, an endogenous circannual rhythm of PRL secretion exists, and that the seasonal cycle of hair growth and moult appears to depend, at least in part, on circulating levels of PRL.     

Relationship between the quality of dietary proteins and the functional status of the adrenal cortex. Variations in the content of transcortion, free corticosterone and total corticosterone in the blood plasma of rats fed with proteins of various quality]

Growing male Wistar rats were used to test in which way variations in the quality of dietary proteins (10% absorbable crude protein; casein supplemented with 4% methionine (K); casein/gelatine (1 : 3); maize gluten supplemented with 4 amino acids; and maize gluten) influenced the binding capacity of blood plasma for corticosterone, and the concentration of corticosteroid-binding globulin ("CBG"), total corticosterone, free corticosterone and albumin-bound corticosterone in blood plasma. The binding capacity of the blood plasma for corticosterone and the total concentration of corticosterone, CBG, free corticosterone and albumin-bound corticosterone were found to rise with the improving quality of the dietary proteins (r = 0.830). A close correlation was found to exist between the concentration of total corticosterone in the blood plasma and CBG. Increases in the total concentration of corticosterone in blood plasma are, in 84% of all cases, brought about by increases in CBG concentrations. In conclusion, reference is made to the significance which such results may have for the control of regulatory processes.     

Intrauterine infusion of BQ-610, an endothelin type A receptor antagonist, delays luteolysis in dairy heifers

Three separate in vivo experiments were conducted to evaluate the putative role of endothelin-1 (ET-1) during luteal regression in heifers. In Experiment 1, a single intraluteal injection of 500 μg BQ-610 [(N,N-hexamethylene) carbamoyl-Leu-d-Trp (CHO)-d-Trp], a highly specific endothelin A (ET_A) receptor antagonist, did not diminish the decline in plasma progesterone following a single exogenous injection of 25 mg prostaglandin F2 alpha (PGF₂) administered at midcycle of the estrous cycle. In Experiment 2, six intrauterine infusions of 500 μg BQ-610 given every 12 h on days 16–18 delayed spontaneous luteolysis, as evidenced by an extended elevation (P = 0.054) of plasma progesterone concentration. In Experiment 3, heifers were administered six intrauterine infusions of BQ-610 or saline on days 16–19, and peripheral blood samples were collected from day 11 to 16 (before infusion), hourly on days 16–19 (during infusion), and on days 20–25 (after infusion). BQ-610 treated heifers had markedly higher (P < 0.0001) levels of plasma progesterone compared with saline controls, and this effect was most notable during the infusion period (treatment by period interaction; P ≤ 0.05). Heifers infused with BQ-610 also had higher progesterone levels on day 21 (treatment by time interaction; P ≤ 0.05). Mean plasma concentrations of 13,14-dihydro-15-keto-PGF₂ (PGFM), the primary metabolite of PGF₂, were measured in the samples collected hourly and were not different (P ≥ 0.05) between treatments. These results indicate that the in vivo antagonism of the ET_A receptor can delay functional luteolysis, and supports the theory that ET-1 regulates luteal function in ruminants.     

Postnatal changes in electromyographic signals during piglet growth, and in relation to muscle fibre types

This study uses non-invasive evoked surface electromyography (SEMG) to investigate postnatal muscle development in pigs, and to assess any correlation between recorded signal parameters and muscle fibre types in two different skeletal muscles. Four litters (n = 43) of Large White × Landrace pigs were used. Evoked SEMG measurements were taken on days 2, 5, 14, 26, 60 and 151 post partum from m. Longissimus dorsi (LD) and on days 14, 26, 60 and 151 post partum from m. Biceps femoris (BF). A third of each litter was slaughtered at days 27, 61 and 153 post partum. Biopsy samples for LD and BF were taken to categorize muscle fibre types. For LD there was a significant increase in compound muscle action potential (CMAP) parameters from day 2 to day 5 post partum, whilst for BF significant increases occurred from days 14 to 26 post partum. Fibre type development in both muscles showed a significant decrease in type IIA fibre number (P < 0.001) and an increase in type IIB fibre number (P < 0.001). Analysis of CMAP parameters in relation to fibre type percentages showed significant positive correlations between percentage of type I fibres in BF and CMAP Area (r = 0.71; P = 0.05), SlopeL (r = 0.79; P = 0.02) and Corr Peak (r = 0.78; P = 0.02) and a negative correlation with SlopeT (r = 0.89; P = 0.003) at day 151. A greater CMAP Area and Corr Peak as well as steeper leading and trailing slopes were on the other hand correlated to fewer type IIB fibres.

It is concluded that 1) changes in the evoked CMAP are muscle-specific during early postnatal development in the pig, and 2) in spite of a small sample size, the correlations between CMAP signal parameters and fibre type percentages warrant further investigation.     

Studies on vertical transmission of Trichinella spp. in experimentally infected ferrets (Mustela putorius furo), foxes (Vulpes vulpes), pigs, guinea pigs and mice

Vertical transmission of Trichinella spiralis was evaluated in ferrets (n = 21), foxes (n = 11), pigs (n = 12), guinea pigs (n = 16), and mice (n = 41). The placental barrier to be crossed by migratory Trichinella larvae varies structurally in different animal species. Ferrets and foxes have an endotheliochorial placenta structure, guinea pigs and mice a haemochorial, and pigs an epitheliochorial placenta. The non-encapsulating Trichinella pseudospiralis larvae have an extended muscle migration prior to entering a muscle cell. To evaluate if T. pseudospiralis was more likely to be transmitted to offspring, an additional group of foxes (n = 11) infected with T. pseudospiralis was included. Two different dose levels were used for ferrets, pigs, guinea pigs, and mice. In pigs and guinea pigs, infection was given at different times of the gestation period. Vertical transmission, measured as recovery of muscle larvae in the offspring, was demonstrated in both ferrets groups, in all four guinea pig groups, and in the high dose mouse group, but not in any fox or pig groups.     

Phenotypic and genotypic traits of Staphylococcus aureus strains isolated from pig carcasses

A total of 142 S. aureus strains isolated from pig carcasses from abattoirs A (n = 98) and B (n = 44) were characterized by phenotypic and genotypic traits. Phenotypically, 96% showed yellow-pigmented colonies, 63% β/δ hemolysis, 85% were egg yolk-positive and 99% were positive for clumping factor/protein A. Only 25% of the strains were resistant to the antimicrobials tested (abattoir A: 19%; abattoir B: 39%), especially to penicillin and ampicillin. None of the strains harbored the mecA gene, which is conserved in methicillin-resistant S. aureus. The biofilm associated genes icaA, icaD and bap were present in 100%, 100% and 0% of the strains. Genes for staphylococcal enterotoxin (SE) were detected in 51% (abattoir A) and 14% of the strains (abattoir B). Among strains harboring SE genes (n = 56), 63%, 31%, 4% and 2% tested positive for seg/sei, seg, sei and sec, respectively. The amplification of the 3′ end of the coagulase gene (coa) yielded amplicons of 400, 436, 602, 682 or 776 bp. Coa restriction profile (CRP) analysis using HaeIII resulted in seven patterns (a–d, e1–e3). CRP (c) was detected most frequently at both abattoirs, whereas CRP (a) was restricted to abattoir A and CRP (e3) to abattoir B. In the slaughter process (abattoir B), (i) two CRPs (b and d) were only found before dehairing/singeing, and (ii) four CRPs (c, e1–e3) were identified throughout the process. The genotyping revealed a remarkable homogeneity in S. aureus strains from the two different abattoirs and the slaughter process stages. These results may be explained by the distribution of a limited number of S. aureus genotypes in the pig population. Moreover, as the predominant CRPs (c, e1–e3) persisted throughout the slaughter process in abattoir B, it may be hypothesized that these types are characterized by colonization advantages.     

Immunoglobulin G concentration and neonatal survival of goat kids delivered in a pen or on open range

To determine whether kids born on open range (subjected to stress due to management practices) had lower IgG concentrations and a higher mortality risk than kids born in a pen, serum IgG concentrations in 63 one-day-old kids were determined. Median Serum IgG concentrations one day after birth tended to be lower (p=0.09) in surviving kids born in the open than that of surviving kids delivered in pen (115 vs. 1119 mg dl⁻¹). However, survival risk (71 and 82% for kids born on the open range and pen, respectively; n=79) were not significantly affected by site of kidding. Median IgG concentration of surviving kids born on a pen was not significantly different (p=0.13) compared to non-surviving kids (1118 vs. 115 mg dl⁻¹). No differences were detected in either median serum IgG levels or death rates between male and female kids. Kids with IgG concentrations <800 mg dl⁻¹ showed lower survival risks than those with higher concentrations. We concluded that typical management practices of kids delivered on open range at a goat operation under extensive conditions in northern Mexico tended to reduced colostrally acquired serum immunoglobulin levels, but had no effect directly on mortality.     

Use of PCR-RFLP to identify Leishmania species in naturally-infected dogs

Tissue imprints on Giemsa stained slides from dogs were used to investigate the presence of Leishmania amastigotes by either optical microscopy (OM) or Polymerase chain reaction (PCR) detection of DNA. Samples from skin, spleen, lymph node, liver and bone marrow from a Leishmaniasis endemic area dogs where Leishmania (Leishmania) chagasi and Leishmania (Viannia) braziliensis are sympatric were studied. Dogs were initially diagnosed by Indirect Immunofluorescence (IIF), as which 39 were IIF positive (≥1:40) and 16 negative. The IIF positive dogs were clinically grouped as symptomatic (n = 15), oligosymptomatic (n = 12) and asymptomatic (n = 12). Although PCR positivity was higher in symptomatic dogs, specially their skin samples, there was no significant difference among clinical groups or organs examined. Ten (62.5%) out of 16 IIF and OM negative animals were positive for PCR in at least one organ. Forty-eight positive PCR amplicons were further submitted to RFLP for Leishmania identification. All dogs were infected with L. (L.) chagasi except one, infected with L. (V.) braziliensis. PCR was more efficient than IIF and OM to diagnose canine visceral Leishmaniasis (CVL), regardless of the organ examined and the clinical form present. The use of PCR together with serology helps determining the extension of sub clinical infection in CVL endemic areas and provides a better estimate of the number of dogs to be targeted for control measures. In conclusion, our data reinforce the need for a specific diagnosis of canine infection in areas where diverse Leishmania species are sympatric and demonstrate that PCR–RFLP can be used to identify Leishmania species in dog tissue imprint stained slides.     

Risk factors for intramammary infections and relationship with somatic-cell counts in Italian dairy goats

Routine examination of milk was performed on five herds of lactating goats in northern Italy as part of a milk quality-monitoring program in the year 2000. As part of the study, aseptic samples of foremilk were collected monthly from both half udders during the entire lactation for 305 goats, resulting in a total of 4571 samples. The samples were tested with cytological and bacteriological analyses to evaluate the relationship between mammary infections and somatic-cell count (SCC; Fossomatic (TM) method). Prevalence of intramammary infection (IMI) was 40.2% (n = 1837) of all udder-half samples examined. The most-prevalent mastitis agents were coagulase-negative Staphylococci (CNS), 80% (n = 1474 udder-half samples); within this group, Staphylococcus epidermidis was the most-prevalent species (38%). Other prevalence were Staphylococcus aureus 6% (n = 112 udder-half samples) and environmental pathogens 14% of infected udder-half samples (n = 251) with a diverse mixture of species, none of which had a frequency of >4%. Enterococcus faecalis was the most-frequently isolated among this group. Neither Salmonella spp. nor Listeria monocytogenes were detected. The risk (sample level) of infection differed across herds, parities, and stage of lactation according to results from logistic multiple regression. Infection was more common among goats in third and fourth parities and during the later stages of lactation. Of the 2734 samples from uninfected udder halves, the mean log₂ SCC was 3.9 cell/ml; of the 1837 bacteriological positive samples, the mean log₂ SCC was 5.6 cell/ml. According to results from a linear mixed model, concentrations of somatic cells tended to increase with increasing age and days in milk and with the presence of bacteria. Infection with S. aureus was associated with the highest SCS.     

Endotoxin induces delayed ovulation following endocrine aberration during the proestrous phase in Holstein heifers

The effect of endotoxin on follicular growth and on secretion of LH, estradiol-17β, progesterone and cortisol during the proestrous phase in cattle was investigated. Holstein heifers were treated with PGF₂ at 11–13 d after ovulation to induce luteolysis. At 42 hr after PGF₂ treatment, heifers were administered either lipopolysaccharide (LPS; Escherichia coli, O111:B4, 5 μg/kg, n = 6) or saline (control; n = 6) by i.v. bolus injection. Ovarian structures were monitored daily by transrectal ultrasonography, and blood samples were collected at various times for hormonal analysis. The duration from PGF₂ treatment to ovulation was significantly longer in the LPS group (8.0 ± 1.3 d) than in the control group (4.2 ± 0.2 d). LPS significantly reduced the pulse frequency of LH for 6 hr after the administration, and increased the mean concentration and pulse amplitude of LH from 3 to 6 hr after the administration. The plasma concentrations of progesterone and cortisol were transiently increased after LPS administration. The plasma concentration of estradiol-17β was significantly decreased at 24 hr after LPS administration compared to that in the controls. Five of six LPS-treated heifers exhibited no preovulatory LH surge until 120 hr after PGF₂ treatment and the remaining heifer exhibited the surge at 108 hr after PGF₂ treatment, while the LH surge was observed at 54–78 hr after PGF₂ treatment in control heifers. These results suggest that endotoxin disrupts progression of the proestrous phase of cattle, interrupting the preovulatory estradiol rise and thus delaying the LH surge and the subsequent ovulation.     

GnIH基因克隆、表达及对幼龄公兔生殖激素的影响

旨在获得兔促性腺激素抑制素(gonadotropin-inhibitory hormone,GnIH)基因，检测其在不同组织和不同发育阶段下丘脑中的表达水平，研究其对幼龄公兔生殖激素分泌的影响。本研究以90日龄健康闽西南黑兔公兔为研究对象，采用RACE技术(rapid-amplification of cDNA ends)从下丘脑组织中克隆出GnIH基因，并对序列进行生物信息学分析；采用实时荧光定量PCR方法(real-time quantitative PCR,qPCR)检测GnIH基因mRNA在90日龄公兔不同组织表达水平(n=5)及11、30、60、90、120、150日龄公兔下丘脑中的表达水平(n=6);连续10 d向80日龄公兔分别注射0、0.5、5和50μg鹌鹑GnIH相关肽(n=10)。第11天早上采集耳静脉血液后，屠宰公兔，采集睾丸组织，分别检测血清中生殖激素浓度(GnRH、FSH、LH、INHB和T)和睾丸组织中睾酮合成相关酶基因(StarR、3β-HSD和p450scc)mRNA的表达水平。结果表明，兔GnIH基因cDNA全长904 bp,包含5′UTR 41 bp...     

Follicular fluid concentrations of free insulin-like growth factor (IGF)-I during follicular development in mares

The objective of the present study was to evaluate changes in concentrations of free insulin-like growth factor (IGF)-I in follicular fluid (FFL) during follicle development in the mare. Mares (n = 14) were classified as either in the follicular phase (n = 8) or luteal phase (n = 6). Follicles (n = 92) were categorized as small (6–15 mm; n = 54), medium (16–25 mm; n = 23) or large (>25 mm; n = 15) and FFL was collected. Free IGF-I levels in FFL in large follicles of follicular phase mares were greater (P < 0.05) than in large follicles of luteal phase mares and small or medium follicles of luteal and follicular phase mares. Free IGF-I concentrations were greater (P < 0.05) in large follicles of luteal phase mares than small but not medium follicles of luteal phase mares. FFL ratio of estradiol:progesterone paralleled changes in free IGF-I. Free IGF-I concentrations were negatively correlated (P < 0.05) with insulin-like growth factor binding protein (IGFBP)-2, -4 and -5 but not IGFBP-3 levels. In addition, free IGF-I concentrations in FFL were positively correlated (P < 0.01) with FFL estradiol, progesterone, androstenedione, estradiol:progesterone ratio, total IGF-I and total IGF-II. We conclude that increases in intrafollicular levels of bioavailable (free) IGF-I are associated with increased steroidogenesis in developing mare follicles.