Serum Lipid Modification Related to Exercise and Polyunsaturated Fatty Acid Supplementation in Jumpers and Thoroughbred Horses

The importance of polyunsaturated fatty acids (PUFAs) within the different biological functions of animals has been widely recognized. In this study, exercise and PUFAs supplementation effects on serum triglycerides, total cholesterol, and nonesterified fatty acids (NEFAs) concentration were evaluated in athletic horses. Two sport horse types (10 Italian saddle jumpers and 10 Thoroughbreds) were equally divided into two groups. Jumpers and Thoroughbred experimental groups (A_J and A_T) received 4-week PUFAs supplementation and control groups (B_J and B_T) received no dietary supplement. Before starting the PUFAs supplementation (T0) and at the end of the experimental period (T4), horses were subjected to simulated events. From each subject, blood samples were collected every 7 days at rest, before and after the first test (T0_R and T0_PE), and before and after the second test (T4_R and T4_PE). Higher triglycerides and NEFA concentrations at T0_PE and T4_PE than T0_R and T4_R in both groups were found as a result of exercise (P < .005), but lower triglycerides and NEFA concentrations at T4_PE in group A_J than group B_J (P < .05) and in group A_T (P < .005) than group B_T were found as a result of PUFAs supplementation. Effects of PUFAs supplementation was highlighted by the statistically significant lower triglycerides and NEFA concentrations found at T4_PE than T0_PE in groups A_J (P < .05) and A_T (triglycerides: P < .05; NEFAs: P < .0001).     

Phylogenetic distribution of virulence genes in Escherichia coli isolated from bovine mastitis in Iran

One hundred and twenty seven Escherichia coli isolates from bovine mastitis were examined to detect the phylogenetic group/subgroups and a selection of virulence associated genes. Forty nine (38.58%) isolates belonged to group B1 the remaining isolates fell into four phylogenetic subgroups: A₀ (18.11%), A₁ (26.77%), D₁ (6.29%) and D₂ (10.23%). None of the isolates belonged to B2 group. Forty seven (37.00%) isolates were positive for at least one virulence gene, among them f17A was the most common gene, found in 20.47% of the isolates. Among the E. coli isolates, 11.81% had iucD, 9.44% f17c-A, 9.44% cnf2, 7.87% f17b-A, 6.29% afaD-8 and afaE-8, 3.14% f17d-A, 0.78% cnf1 and 0.78% clpG genes. All of the detected virulence genes were present alone or in combination with each other except clpG and f17d-A genes that were only found alone. None of the isolates contained the genes for F17a-A, intimin, P or S fimbriae.     

Further characterization of diabetes mellitus and body weight loss in males of the congenic mouse strain DDD.Cg-Ay

The A^y allele at the agouti locus causes obesity and promotes linear growth in mice. However, body weight gain stops between 16 and 17 weeks after birth, and then, body weight decreases gradually in DDD.Cg-A^y male mice. Body weight loss is a consequence of diabetes mellitus, which is genetically controlled mainly by a quantitative trait locus (QTL) on chromosome 4. This study aimed to further characterize diabetes mellitus and body weight loss in DDD.Cg-A^y males. The number of β-cells was markedly reduced, and plasma insulin levels were very low in the DDD.Cg-A^y males. Using a backcross progeny of DDD × (B6 × DDD.Cg-A^y) F₁-A^y, we identified one significant QTL for plasma insulin levels on distal chromosome 4, which was coincidental with QTL for hyperglycemia and lower body weight. The DDD allele was associated with decreased plasma insulin levels. When the DDD.Cg-A^y males were housed under three different housing conditions [group housing (4 or 5 DDD.Cg-A^y and DDD males), individual housing (single DDD.Cg-A^y male) and single male housing with females (single DDD.Cg-A^y male with DDD.Cg-A^y or DDD females)], diabetes mellitus and body weight loss were most severely expressed in individually housed mice. Thus, the severity of diabetes and body weight loss in the DDD.Cg-A^y males was strongly influenced by the housing conditions. These results demonstrate that both genetic and nongenetic environmental factors are involved in the development of diabetes mellitus and body weight loss in the DDD.Cg-A^y males.     

Distinct adiponectin profiles might contribute to differences in susceptibility to type 2 diabetes in dogs and humans

Dogs develop obesity-associated insulin resistance but not type 2 diabetes mellitus. Low adiponectin is associated with progression to type 2 diabetes in obese humans. The aims of this study were to compare total and high molecular weight (HMW) adiponectin and the ratio of HMW to total adiponectin (S_A) between dogs and humans and to examine whether total or HMW adiponectin or both are associated with insulin resistance in naturally occurring obese dogs. We compared adiponectin profiles between 10 lean dogs and 10 lean humans and between 6 lean dogs and 6 age- and sex-matched, client-owned obese dogs. Total adiponectin was measured with assays validated in each species. We measured S_A with velocity centrifugation on sucrose gradients. The effect of total and HMW adiponectin concentrations on MINMOD-estimated insulin sensitivity was assessed with linear regression. Lean dogs had total and HMW adiponectin concentrations three to four times higher than lean humans (total: dogs 32 ± 5.6 mg/L, humans 10 ± 1.3 mg/L, P<0.001; HMW: dogs 25 ± 4.5 mg/L, humans 6 ± 1.3 mg/L, P<0.001) and a higher S_A (dogs: 0.78 ± 0.05; humans: 0.54 ± 0.08, P = 0.002). Adiponectin concentrations and S_A were not lower in obese dogs (0.76 ± 0.05 in both groups; P=1). Total adiponectin, HMW adiponectin, and S_A were not associated with insulin sensitivity in dogs. We propose that differences in adiponectin profiles between humans and dogs might contribute to the propensity of humans but not dogs to develop type 2 diabetes. Dogs with chronic, naturally occurring obesity do not have selectively reduced HMW adiponectin, and adiponectin does not appear to be important in the development of canine obesity-associated insulin resistance.     

Effect of the Y chromosome on testis weight in mice

We investigated the effect of the Y chromosome on testis weight in (B6.Cg-A^y × Y-consomic mouse strain) F₁ male mice. We obtained the following results: (1) Mice with the Mus musculus domesticus-type Y chromosome had significantly heavier testis than those with the M. m. musculus-type Y chromosome. (2) Variations in Usp9y and the number of CAG repeats in Sry were significantly associated with testes weight. The A^y allele was correlated with a reduced testis weight, and the extent of this reduction was significantly associated with a CAG repeat number polymorphism in Sry. These results suggest that Y chromosome genes not only influence testis weight but also modify the effect of the A^y allele in mediating this phenomenon.     

Generation of transgenic corn-derived Actinobacillus pleuropneumoniae ApxIIA fused with the cholera toxin B subunit as a vaccine candidate

Corn, one of the most important forage crops worldwide, has proven to be a useful expression vehicle due to the availability of established transformation procedures for this well-studied plant. The exotoxin Apx, a major virulence factor, is recognized as a common antigen of Actinobacillus (A.) pleuropneumoniae, the causative agent of porcine pleuropneumonia. In this study, a cholera toxin B (CTB)-ApxIIA#5 fusion protein and full-size ApxIIA expressed in corn seed, as a subunit vaccine candidate, were observed to induce Apx-specific immune responses in mice. These results suggest that transgenic corn-derived ApxIIA and CTB-ApxIIA#5 proteins are potential vaccine candidates against A. pleuropneumoniae infection.     

GABA(A) receptor antagonism increases NMDA receptor inhibition by isoflurane at a minimum alveolar concentration

ObjectiveAt the minimum alveolar concentration (MAC), isoflurane potentiates GABA_A receptor currents and inhibits NMDA receptor currents, and these actions may be important for producing anesthesia. However, isoflurane modulates GABA_A receptors more potently than NMDA receptors. The objective of this study was to test whether isoflurane would function as a more potent NMDA receptor antagonist if its efficacy at GABA_A receptors was decreased.Study designProspective experimental study.AnimalsFourteen 10-week-old male Sprague–Dawley rats weighing 269 ± 12 g.MethodsIndwelling lumbar subarachnoid catheters were surgically placed in isoflurane-anesthetized rats. Two days later, the rats were anesthetized with isoflurane, and artificial CSF containing either 0 or 1 mg kg^?1 picrotoxin, a GABA_A receptor antagonist, was infused intrathecally at 1 μL minute^?1. The baseline isoflurane MAC was then determined using a standard tail clamp technique. MK801 (dizocilpine), an NMDA receptor antagonist, was then administered intravenously at 0.5 mg kg^?1. Isoflurane MAC was re-measured.ResultsPicrotoxin increased isoflurane MAC by 16% compared to controls. MK801 significantly decreased isoflurane MAC by 0.72% of an atmosphere in controls versus 0.47% of an atmosphere in rats receiving intrathecal picrotoxin.Conclusions and clinical relevanceA smaller MK801 MAC-sparing effect in the picrotoxin group is consistent with greater NMDA antagonism by isoflurane in these animals, since it suggests that fewer NMDA receptors are available upon which MK801 could act to decrease isoflurane MAC. Decreasing isoflurane GABA_A potentiation increases isoflurane NMDA antagonism at MAC. Hence, the magnitude of an anesthetic effect on a given channel or receptor at MAC may depend upon effects at other receptors.     

The bispectral index during recovery from halothane and sevoflurane anaesthesia in horses

ObjectiveTo record the bispectral index (BIS) when horses moved during either halothane or sevoflurane anaesthesia and when they made volitional movements during recovery from these anaesthetics.Study designRandomized prospective clinical study.AnimalsTwenty-five client-owned horses undergoing surgery aged 8.8 (± 5.3; 1–19) years (mean ± SD; range).MethodsBaseline BIS values were recorded before pre-anaesthetic medication (BIS_B) and during anaesthesia (BIS_A) maintained with halothane (group H; n = 12) or sevoflurane (group S; n =13) at approximately 0.8–0.9 × minimum alveolar concentrations (MAC). Bispectral indices were recorded during the surgery when unexpected movement occurred (BIS_MA), during recovery when the first movement convincingly associated with consciousness was observed (BIS_M1) and once sternal recumbency was achieved (BIS_ST).ResultsNo significant difference in BIS_M1 was found between halothane- (85 ± 7; 75–93) and sevoflurane- (87 ± 10; 70–98) anaesthetized horses although BIS_A was significantly (p = 0.0002) lower in group S (62 ± 7; 53–72) than group H (74 ± 7; 60–84). Differences between BIS_M1 and BIS_A were significant in sevoflurane (p = 0.00001) and halothane recipients (p = 0.002) but were greater in group S (25 ± 9; 4–38) compared with group H (12 ± 10; ?9–25). In six of eight horses, BIS_MA values ranged between those recorded during anaesthesia and at first movement.Conclusions and clinical relevanceBispectral indices appear to approximate levels of unconsciousness, suggesting that monitoring the BIS may assist equine anaesthesia. However, it does not predict intra-operative movement.     

Comparison of pain on injection during induction of anaesthesia with alfaxalone and two formulations of propofol in dogs

ObjectiveTo compare the incidence of pain during injection of three intravenous induction agents in dogs.Study designProspective, crossover, randomized, blinded, clinical study.AnimalsThirty dogs requiring anaesthesia for radiotherapy.MethodsDogs were anaesthetized on three occasions at weekly intervals. An IV cephalic catheter was placed, flushed with saline and alfentanil 0.01 mg kg^?1 and atropine 0.02 mg kg^?1 administered. After 30 seconds either: propofol lipid macroemulsion (Drug_P), propofol lipid-free microemulsion (Drug_PC) or alfaxalone (Drug_A) was administered over 60 seconds. Each induction agent was administered once to each dog. Induction was recorded by video and reviewed by an assessor, unaware of treatment. Catheter placement (number of attempts, site, size and recent vein use) were recorded. Behavioural changes associated with pain or excitation, were recorded. Severity of pain on injection was recorded (mild, moderate or severe pain). Incidence of pain was analysed using logistic regression, excitation using McNemar's test (p < 0.05) and association of pain with induction agent and catheter placement using the Akaike Information Criterion (AIC).ResultsNo dogs reacted to saline or Drug_A, thus Drug_A was excluded from analysis. Pain on injection occurred in six dogs (20%) with Drug_PC and one dog (3.3%) with Drug_P. Pain was severe in four dogs with Drug_PC. Drug_P resulted in a trend for reduced risk of pain compared to Drug_PC (p = 0.076, odds ratio [confidence intervals] 0.14 [0.027–0.86]). Both propofol formulations resulted in greater risk of excitation than Drug_A (p = 0.0003, odds ratio 4.5 [1.86–10.90]). Induction agent was associated with pain, whilst catheter placement was not. One dog developed facial oedema and one other dog skin necrosis adjacent to the catheter site following Drug_PC. The study was terminated early due to ethical concerns about the severity of reactions with Drug_PC.Conclusions and Clinical relevanceDrug_PC was associated with clinically relevant moderate to severe pain behaviour whilst Drug_A and Drug_P were not.     

饲料添加苜蓿草粉对刺参幼参生长、体成分及免疫酶的影响

为了探究紫花苜蓿草粉的适宜添加比例,研究了苜蓿草粉的添加水平依次为0(对照),5%(A₁),10%(A₂),15%(A₃)和20%(A₄) 的5种饲料对刺参幼参(3.16±0.66) g生长、体成分和免疫酶活性的影响。结果显示,刺参特定生长率(SGR)的最大值(0.73%)和饵料系数(FCR)的最小值都出现在A₂组。A₂组刺参的粗蛋白含量最高(52.10%),显著高于A₁与A₄组(P<0.05)。A₄组刺参的过氧化氢酶(CAT)活力值最高(142.28 U/mL),显著高于A₀、A₁与A₂组(P<0.05);A₃组刺参的超氧化物歧化酶(SOD)活力值最高(136.58 U/mL),显著高于A₀组(P<0.05); A₃组刺参的溶菌酶(LZM)含量最高(1.92 μg/mL),显著高于A₀、A₁与A₂组(P<0.05)。结果表明,当苜蓿草粉的添加比例为10%左右时,实验刺参的养殖效果最佳。     

The Activity and Localization of 3β-hydroxysteroid Dehydrogenase/Δ5-Δ4 Isomerase and Release of Androstenedione and Progesterone by Uterine Tissues During Early Pregnancy and the Estrous Cycle in Pigs

Steroid hormones are produced by the porcine uterus. We hypothesized that the uterus in pigs possesses active 3β-hydroxysteroid dehydrogenase/Δ⁵-Δ⁴ isomerase (3β-HSD) responsible for progesterone and androstenedione production, that uterine steroids may supplement the amount of steroid hormones produced by embryos and corpus luteum and that these steroids are necessary for maintenance of pregnancy. In this study, we examined 1) endometrial and myometrial expression of 3β-HSD mRNA, 2) uterine 3β-HSD protein activity and 3) in vitro production of A₄ and P₄ by uterine slices harvested from pigs on days 10 to 11, 12 to 13 and 15 to 16 of pregnancy and the estrous cycle. The expression of 3β-HSD and the presence and activity of 3β-HSD protein were different in the endometrium and the myometrium during the examined periods of pregnancy and the estrous cycle. Production of A₄ by the endometrium and myometrium was highest on days 12 to 13 of pregnancy and the estrous cycle. Endometrial secretion of P₄ did not differ in the course of early pregnancy and on the respective days of the estrous cycle. The gravid myometrium was the highest source of P₄ in pregnant pigs on days 12 to 13. The release of P₄ by the cyclic myometrium rose during the examined days of the estrous cycle. The steroidogenic activity of the uterus, as described in this study, may support early pregnancy or the luteal phase of the estrous cycle in pigs.     

Effects of subcutaneous alfaxalone alone and in combination with dexmedetomidine and buprenorphine in guinea pigs (Cavia porcellus)

Objective

To characterize alfaxalone administered subcutaneously (SC) in guinea pigs, both alone and in combination with dexmedetomidine and buprenorphine.

Serotonin markers show altered transcription levels in an experimental pig model of mitral regurgitation

Serotonin (5-hydroxytryptamine, 5-HT) signalling is implicated in the pathogenesis of myxomatous mitral valve disease (MMVD) through 5-HT_1B receptor (R), 5-HT_2AR and 5-HT_2BR-induced myxomatous pathology. Based on increased tryptophan hydroxylase-1 (TPH-1) and decreased serotonin re-uptake transporter (SERT) in MMVD-affected valves, increased valvular 5-HT synthesis and decreased clearance have been suggested. It remains unknown how haemodynamic changes associated with mitral regurgitation (MR) affect 5-HT markers in the mitral valve, myocardium and circulation. Twenty-eight pigs underwent surgically induced MR or sham-operation, resulting in three MR groups: control (CON, n = 12), mild MR (mMR, n = 10) and severe MR (sMR, n = 6). The gene expression levels of 5-HT_1BR, 5-HT_2AR, 5-HT_2BR, SERT and TPH-1 were analysed using quantitative PCR (qPCR) in the mitral valve (MV), anterior papillary muscle (AP) and left ventricle (LV). MV 5-HT_2BR was also analysed with immunohistochemistry (IHC) in relation to histological lesions and valvular myofibroblasts. All 5-HTR mRNAs were up-regulated in MV compared to AP and LV (P <0.01). In contrast, SERT and TPH-1 were up-regulated in AP and LV compared to MV (P <0.05). In MV, mRNA levels were increased for 5-HT_2BR (P = 0.02) and decreased for SERT (P = 0.03) in sMR vs. CON. There were no group differences in 5-HT_2BR staining (IHC) but co-localisation was found with α-SMA-positive cells in 91% of all valves and with 33% of histological lesions. In LV, 5-HT_1BR mRNA levels were increased in sMR vs. CON (P = 0.01). In conclusion, these data suggest that MR may affect mRNA expression of valvular 5-HT_2BR and SERT, and left ventricular 5-HT_1BR in some pigs.     

Co-expression of the Bcl-xL antiapoptotic protein enhances the induction of Th1-like immune responses in mice immunized with DNA vaccines encoding FMDV B and T cell epitopes

Foot-and-mouth disease (FMD) is one of the most devastating animal diseases, affecting all cloven-hoofed domestic and wild animal species. Previous studies from our group using DNA vaccines encoding FMD virus (FMDV) B and T cell epitopes targeted to antigen presenting cells, allowed demonstrating total protection from FMDV homologous challenge in those animals efficiently primed for both humoral and cellular specific responses (Borrego et al. Antivir Res 92:359-363, 2011). In this study, a new DNA vaccine prototype expected to induce stronger and cross-reactive immune responses against FMDV which was designed by making two main modifications: i) adding a new B-cell epitope from the O-serotype to the B and T-cell epitopes from the C-serotype and ii) using a dual promoter plasmid that allowed inserting a new cistron encoding the anti-apoptotic Bcl-xL gene under the control of the internal ribosomal entry site (IRES) of encephalomyocarditis virus aiming to increase and optimize the antigen presentation of the encoded FMDV epitopes after in vivo immunization. In vitro studies showed that Bcl-xL significantly prolonged the survival of DNA transfected cells (p?<?0.001). Accordingly, vaccination of Swiss out-bred mice with the dual promoter plasmid increased the total IgG responses induced against each of the FMDV epitopes however no significant differences observed between groups. The humoral immune response was polarized through IgG2a in all vaccination groups (p?<?0.05); except peptide T_3A; in correspondence with the Th1-like response observed, a clear bias towards the induction of specific IFN-γ secreting CD4⁺ and CD8⁺ T cell responses was also observed, being significantly higher (p?<?0.05) in the group of mice immunized with the plasmid co-expressing Bcl-xL and the FMDV B and T cell epitopes.     

Evaluation of the reliability of pulse oximetry,at different attachment sites,to detect hypoxaemia in immobilized impala (Aepyceros melampus)

ObjectiveEvaluation of the reliability of pulse oximetry at four different attachment sites compared to haemoglobin oxygen saturation measured by a co-oximeter and calculated by a blood gas analyser in immobilized impala.Study designRandomized crossover study.AnimalsA total of 16 female impala.MethodsImpala were immobilized with etorphine or thiafentanil alone, or etorphine in combination with a novel drug. Once immobilized, arterial blood samples were collected at 5 minute intervals for 30 minutes. Then oxygen was insufflated (5 L minute⁻¹) intranasally at 40 minutes and additional samples were collected. A blood gas analyser was used to measure the arterial partial pressure of oxygen and calculate the oxygen haemoglobin saturation (cSaO₂); a co-oximeter was used to measure the oxygen haemoglobin saturation (SaO₂) in arterial blood. Pulse oximeter probes were attached: under the tail, to the pinna (ear) and buccal mucosa (cheek) and inside the rectum. Pulse oximeter readings [peripheral oxygen haemoglobin saturation (SpO₂) and pulse quality] were recorded at each site and compared with SaO₂ and cSaO₂ using Bland-Altman and accuracy of the area root mean squares (A_rms) methods to determine the efficacy. P value < 0.05 was considered significant.ResultsPulse quality was ‘good’ at each attachment site. SpO₂ measured under the tail was accurate and precise but only when SaO₂ values were above 90% (bias = 3, precision = 3, A_rms = 4). The ear, cheek and rectal probes failed to give accurate or precise readings (ear: bias = −4, precision = 14, A_rms = 15; cheek: bias = 12, precision = 11, A_rms = 16; and rectum: bias = 5, precision = 12, A_rms = 13).Conclusions and clinical relevanceIn order to obtain accurate and precise pulse oximetry readings in immobilized impala, probes must be placed under the tail and SaO₂ must be above 90%. Since SaO₂ values are usually low in immobilized impala, pulse oximeter readings should be interpreted with caution.     

The α2A‐adrenoceptor subtype plays a key role in the analgesic and sedative effects of xylazine

Xylazine, the classical α₂‐adrenoceptor (α₂‐AR) agonist, is still used as an analgesic and sedative in veterinary medicine, despite its low potency and affinity for α₂‐ARs. Previous pharmacological studies suggested that the α_2A‐AR subtype plays a role in mediating the clinical effects of xylazine; however, these studies were hampered by the poor subtype‐selectivity of the antagonists used and a lack of knowledge of their bioavailability in vivo. Here, we attempted to elucidate the role of the α_2A‐AR subtype in mediating the clinical effects of xylazine by comparing the analgesic and sedative effects of this drug in wild‐type mice with those in α_2A‐AR functional knockout mice using the hot‐plate and open field tests, respectively. Hippocampal noradrenaline turnover in both mice was also measured to evaluate the contribution of α_2A‐AR subtype to the inhibitory effect of xylazine on presynaptic noradrenaline release. In wild‐type mice, xylazine (10 or 30 mg/kg) increased the hot‐plate latency. Furthermore, xylazine (3 or 10 mg/kg) inhibited the open field locomotor activity and decreased hippocampal noradrenaline turnover. By contrast, all of these effects were abolished in α_2A‐AR functional knockout mice. These results indicate that the α_2A‐AR subtype is mainly responsible for the clinical effects of xylazine.     

Enhanced IL-6 transcriptional response to adenosine receptor ligands in horses with lower airway inflammation

Reasons for performing study: Accumulation of extracellular adenosine has been closely associated with human asthmatic responses. However, the relevance of adenosine signalling in equine airways has not previously been investigated. Objectives: To determine the expression of adenosine receptors (AR) in bronchoalveolar lavage (BAL) cells and assess the reactivity of these cells to AR ligands ex vivo, employing IL‐6 as readout of adenosinergic inflammatory signalling. Methods: Eight horses with varying degrees of lower airway inflammation and 10 healthy controls were analysed. Expression of AR‐subtypes in each BAL sample was determined by quantitative RT‐PCR and compared to that in 13 other tissues. Bronchoalveolar lavage cells were stimulated either with the adenosine analogue NECA, CGS‐21680 (A_2AAR selective agonist) or with a combination of NECA and SCH‐58261 (A_2AAR antagonist) and IL‐6 expression assessed. Results: Bronchoalveolar lavage cells predominantly expressed A_2BAR, with lower A_2AAR levels and marginal A₃AR expression; A₁AR was not detected. This pattern was similar to that of PBMCs but different from the other tissues tested. No significant differences in AR expression in BAL cells from both groups were detected, although a trend for decreased A_2BAR in airway‐compromised horses was observed. Treatment of BAL cells with the nonselective agonist NECA upregulated IL‐6 expression in cells from airway‐compromised horses, but levels remained unchanged in control animals. Furthermore, blockage of A_2AAR with SCH‐58261 enhanced IL‐6 mRNA induction by NECA in both groups, with higher levels in airway‐compromised horses; the amplitude of this response correlated with neutrophil count. Conclusions: These results demonstrate the presence of an adenosine/IL‐6 inflammatory axis in the bronchoalveolar milieu of airway‐compromised horses. While A_2BAR is the predominant proinflammatory AR subtype expressed, A_2AAR appears to modulate inflammatory signalling (IL‐6 expression) by adenosine. Potential relevance: This study supports selective AR targeting as a potential therapeutic approach for the modulation of inflammation in the equine lower respiratory tract.     

Detection of antibodies to Cooperia ssp. and Ostertagia ssp. in calves with the enzyme linked immunosorbent assay (ELISA)

The ELISA technique was found to be very useful for the detection and monitoring of anti-Cooperia and anti-Ostertagia antibodies in calves. Six differens antigens were used; saline extracts from third- and fourth-stage larvae and from adult worms of both genera.Some degree of genus specificity was found when using L₄ or adult antigens but not when L₃ antigens were used. Stage-specificity could be observed for Cooperia L₄ antigen for a limited period after primary single infection. These findings were supported by the results of immuno-electrophoresis.     

Studies on Erysipelothrix Insidiosa S. Rhusiopathiae: 2. Serological Study

Sixty-two strains of E. insidiosa isolated from joints or regional lymph nodes of pigs with arthritis were examined for their antigenic characteristics. HCl and CH₃COOH extracts were made from all strains and precipitation tests performed with these extracts and rabbit anti-sera for types A₁, A₂, B₁, B₂, C, D, E, and G.The type distribution of the strains was as follows: 1 belonged in type A, 46 in type B, while 15, which did not contain demonstrable acid-soluble antigen, were designated N strains. The 46 type B strains could be divided into 4 subtypes on the basis of the patterns of reaction between, on the one hand HCl and CH₃COOH extracts, and, on the other hand, B₁ and B₂ sera. Thirty-two strains reacted with both B₁ and B₂ sera in CH₃COOH extracts, but not in HCl extracts; these strains were referred to a new subtype, B₃. Nine strains, which reacted with B₁ serum in HCl and CH₃COOH extracts and with B₂ serum in CH₃COOH extracts only, were referred to subtype B₁. Four strains that reacted with B₁ and B₂ sera in both extracts were referred to subtype B₂. The only representative of the 4th subtype was an interesting strain which in previous studies had been shown to differ from the rest of the strains in other ways, too. This strain reacted with both B₁ and B₂ sera, but only in HCl extract.     

Identification of stable normalization genes for quantitative real-time PCR in porcine articular cartilage

Background

Expression levels for genes of interest must be normalized with an appropriate reference, or housekeeping gene, to make accurate comparisons of quantitative real-time PCR results. The purpose of this study was to identify the most stable housekeeping genes in porcine articular cartilage subjected to a mechanical injury from a panel of 10 candidate genes.

Results

Ten candidate housekeeping genes were evaluated in three different treatment groups of mechanically impacted porcine articular cartilage. The genes evaluated were: beta actin, beta-2-microglobulin, glyceraldehyde-3-phosphate dehydrogenase, hydroxymethylbilane synthase, hypoxanthine phosphoribosyl transferase, peptidylprolyl isomerase A (cyclophilin A), ribosomal protein L4, succinate dehydrogenase flavoprotein subunit A, TATA box binding protein, and tyrosine 3-monooxygenase/tryptophan 5-monooxygenase activation protein—zeta polypeptide. The stability of the genes was measured using geNorm, BestKeeper, and NormFinder software. The four most stable genes measured via geNorm were (most to least stable) succinate dehydrogenase flavoprotein, subunit A, peptidylprolyl isomerase A, glyceraldehyde-3-phosphate dehydrogenase, beta actin; the four most stable genes measured via BestKeeper were glyceraldehyde-3-phosphate dehydrogenase, peptidylprolyl isomerase A, beta actin, succinate dehydrogenase flavoprotein, subunit A; and the four most stable genes measured via NormFinder were peptidylprolyl isomerase A, succinate dehydrogenase flavoprotein, subunit A, glyceraldehyde-3-phosphate dehydrogenase, beta actin.

Conclusions

BestKeeper, geNorm, and NormFinder all generated similar results for the most stable genes in porcine articular cartilage. The use of these appropriate reference genes will facilitate accurate gene expression studies of porcine articular cartilage and suggest appropriate housekeeping genes for articular cartilage studies in other species.