Differences in calpain system,desmin degradation and water holding capacity between commercial Meishan and Duroc × Landrace × Yorkshire crossbred pork

The objective of this study was to examine the differences in calpain system, desmin degradation, pH values and water holding capacity (WHC) between muscles of commercial Meishan and Duroc × Landrace × Yorkshire crossbred pigs. Meishan pork presented better WHC evidenced by lower purge loss at days 1 and 3 and less centrifugation loss at day 1 post mortem (P < 0.05). pH values at 45 min post mortem in Meishan pork were significantly higher compared to crossbred pork (P < 0.05). Calpain‐1 messenger RNA (mRNA) expression was lower in Meishan pork compared to that from crossbred pork (P < 0.05). Additionally, calpain‐1 activity, the ratio of calpain‐1 to calpastatin activity and desmin degradation were lower in Meishan pork compared to those from crossbred pork samples (P < 0.05). The results indicate that the calpain system including mRNA expression and activity were different between commercial Meishan and crossbred pork resulting in difference in the degree of desmin degradation during post mortem aging. pH values at 45 min and 24 h post mortem rather than calpain activity and desmin degradation could explain the higher water holding capacity in commercial Meishan pork.     

Influence of Bacillus subtilis GCB‐13‐001 on growth performance,nutrient digestibility,blood characteristics,faecal microbiota and faecal score in weanling pigs

The present study investigated the influence of Bacillus subtilis GCB‐13‐001 on growth performance, nutrient digestibility, blood characteristics, faecal microbiota and faecal score in weanling pigs. A total of 120 weaning pigs [(Landrace × Yorkshire) × Duroc; 7.73 ± 0.75 kg (28 days of age)] were randomly allotted into three treatments according to their initial body weight (BW) and gender in a 6‐week experiment. There were 8 replication pens in each treatment, with five pigs/pen. Dietary treatment groups were as follows: (a) basal diet (CON), (b) CON + 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁸ CFU/kg (T1) and (c) CON + 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁹ CFU/kg (T2). Days 1 to 7, the BW and ADG with T2 treatment were higher (p < .05) than CON treatment, as well as F:G showed trends in linear reduction (p < .1). Days 8 to 21, the BW and ADG were improved (p < .05) in pigs offered T1 and T2 diets compared with CON diet. Days 22 to 42, BW and ADG were higher (p < .05) in pigs fed T2 diet than CON and T1 diets, and the pigs fed T1 diet had higher BW than CON treatment. Overall, the ADG with the T2 treatment was higher (p < .05) than that with the T1 and CON treatments, and pigs offered T1 treatment had higher (p < .05) ADG than CON treatment. Moreover, F:G ratio were significantly decreased (p < .05) by T2 treatment compared with CON treatment. The faecal Lactobacillus counts were improved, and E. coli counts were reduced (p < .05) in pigs fed T2 diet compared with CON at the end of the experiment. In conclusion, supplementation of 0.1% Bacillus subtilis GCB‐13‐001 1 × 10⁹ CFU/kg has shown a beneficial effect in improving BW, increase ADG, decrease F:G ratio.     

Feline platelet counting with prostaglandin E1 on the Sysmex XT‐2000iV

Background: The large size of many feline platelets and the high frequency of platelet aggregation often results in falsely low platelet counts in this species. A combination of optical platelet counting to detect even large platelets and the use of prostaglandin E1 (PGE1) to inhibit platelet clumping may increase the accuracy of feline platelet counting. Objective: The objective of this study was to compare platelet counts in feline whole blood samples with and without the addition of PGE1 and using different analytical methods in a clinical setting. Methods: Platelet counts were determined in 10 feline patients in a referral veterinary hospital using 2 sample types (EDTA, EDTA with PGE1) and 2 methods of analysis (optical counting [PLT‐O] and impedance counting [PLT‐I]) on the Sysmex XT 2000 iV analyzer. Results: All PGE1–PLT‐O samples had platelet counts of >200 × 10⁹/L. Mean platelet count using PGE1–PLT‐O (410,256±178 × 10⁹/L) was significantly higher (P<.03) compared with PGE1–PLT‐I (256±113 × 10⁹/L), EDTA–PLT‐O (238±107 × 10⁹/L), and EDTA–PLT‐I (142±84 × 10⁹/L) methods. Depending on the method, platelet counts in 2 to 7 of 10 cats were <200 × 10⁹/L when PGE1‐PLT‐O was not used. A slightly increased platelet count in response to treatment of a feline patient with thrombocytopenia would have been missed without use of PGE1–PLT‐O. Conclusions: Using PLT‐O analysis on EDTA samples containing PGE1 provides higher, and therefore likely more accurate, feline platelet counts in a clinical setting.     

Morphology and quantification of sheep pineal glands at pre‐pubertal,pubertal and post‐pubertal periods

The pineal gland is a neuroendocrine organ associated with photoperiodic regulation in mammals. The aim of this study was to evaluate the pineal gland at the pre‐pubertal, pubertal and post‐pubertal periods by means of morphology and stereology. The study examined at total of 24 ovine pineal glands collected from healthy female Akkaraman breed. Thick sections (40 μm) were cut and treated with synaptophysin. Following each thick section, six consecutive sections at a thickness of 5 μm were cut. Each thin section was stained with one of the following dyes: Crossman’s modified triple dye, glial fibrillary acidic protein (GFAP), melatonin marker, periodic acid–Schiff, Von Kossa and AgNOR. The pineal gland volume was measured using Cavalieri’s method. The optical fractionator was used to estimate the total number of pinealocytes. The percentage of parenchyma and connective tissue and degree of vascularization were estimated by the area fraction fractionator method. The pineal gland volumes in the pre‐pubertal, pubertal and post‐pubertal groups were 7.53 ± 1.715 mm³, 11.20 ± 1.336 mm³ and 17.75 ± 1.188 mm³, respectively (p < .5). The number of pinealocytes in the pre‐pubertal, pubertal and post‐pubertal groups was 3,244,000 ± 228,076, 4,438,000 ± 243,610, 7,381,766 ± 406,223, respectively (p < .05). The glands of the post‐pubertal group contained the highest amount of connective tissue (11.49 ± 2.103%; p < .5) and the largest GFAP staining area (p < .05). The melatonin staining density was the highest in the pubertal group. The density of lipofuscin staining was higher in the pubertal and post‐pubertal groups.     

Doxorubicin area under the curve is an important predictor of neutropenia in dogs with naturally occurring cancers

Doxorubicin (DOX) area‐under‐the‐curve (AUC) was calculated for 40 dogs with spontaneously occurring cancers using a previously validated limited‐sampling approach. All dogs were administered a dose of 30 mg/m² by intravenous infusion and serum samples were collected at 5, 45 and 60 minutes post‐infusion. DOX and its major metabolite, doxorubicinol (doxol), were quantified in serum samples using high‐performance liquid chromatography tandem‐mass spectrometry. Wide interpatient variability was observed in the predicted DOX AUC with a coefficient of variation of 34%. A significant relationship was found between DOX AUC and absolute white blood cell count (P = 0.003), absolute neutrophil count (ANC; P = 0.002) and surviving fraction of neutrophils (P = 0.03) approximately 1 week after dosing (nadir). No changes in other hematologic parameters (red blood cells, platelets, lymphocytes, haemoglobin) were found to correlate with DOX AUC. The absolute dose (mg) and the dose per unit body weight (mg/kg) were not significantly correlated with nadir ANC. No relationships were found between maximum serum doxol concentration and myelosuppression. Baseline ANC was also significantly correlated to nadir ANC and a model was constructed using baseline ANC and DOX AUC that significantly described the nadir ANC. These findings demonstrate the important relationship between systemic DOX exposure and degree of neutropenia in dogs, and suggest a potential for individualized, pharmacokinetically‐guided DOX dosing in dogs.     

The Effect of Pasteurella haemolytica A2 Infection on Phagocytosis Efficiency of Caprine Broncho‐Alveolar Macrophages

An experiment was designed to study the in vivo effect of Pasteurella haemolytica A2 infection on the phagocytosis activity of caprine broncho‐alveolar macrophages and the extent of pneumonic lesions. Twelve healthy local Kacang goats, about 7 months of age, were divided into two groups of six. Goats in group 1 were inoculated intratracheally with 4 ml inoculum containing 2.8 × 10⁹ colony‐forming units (CFU)/ml of Staphylococcus aureus. Goats in group 2 were inoculated intratracheally with 4 ml of inoculum containing 9.5 × 10⁸ CFU/ml of Pasteurella haemolytica A2 isolated earlier from pneumonic lungs of goat. At intervals of 3 and 7 days post‐challenge five goats from each group were killed and the lungs were washed with sterile phosphate‐buffered saline. Smears were prepared from the lung washing fluid and the number of macrophages with phagocytic activity was determined. At day 3 post‐infection, goats of both groups showed a similar pattern of pneumonic lesion. The lung washing fluid of goats in group 2 was found to contain numerous neutrophils and macrophages. Goats in group 2 showed significantly (P < 0.05) higher extent of lung lesions than group 1. Similarly, the average extent of lung lesions was significantly (P < 0.05) more severe in group 2 at day 7 post‐infection. The lung washing fluid contained mostly macrophages. The phagocytic activity following S. aureus infection was more efficient and significantly (P < 0.01) higher compared with infection by P. haemolytica A2. There were weak correlations between the extent of pneumonic lesion and the phagocytic activity. Thus, goats with poor phagocytic activity were likely to develop more extensive lung lesions.     

The Post‐Cervical Insemination does not Impair the Reproductive Performance of Primiparous Sows

The study evaluated the reproductive performance of primiparous sows submitted to post‐cervical insemination (PCAI) compared with cervical artificial insemination (CAI). Difficulty with catheter introduction, the occurrence of bleeding or semen backflow during insemination, and volume and sperm cell backflow up to 60 min after insemination were also evaluated. Sows were homogenously distributed, according to body weight loss in lactation, lactation length, weaned piglets, weaning‐to‐oestrus interval and total born in previous farrowing, in two treatments: PCAI (n = 165) with 1.5 × 10⁹ sperm cells in 45 ml (2.4 ± 0.04 doses per sow) and CAI (n = 165) with 3 × 10⁹ sperm cells in 90 ml (2.5 ± 0.04 doses per sow). During PCAI, sows were inseminated in the absence of boars. Transabdominal real‐time ultrasonography was performed at oestrus onset, immediately before the first insemination and at 24 h after last insemination. There was no difference (P > 0.05) between treatments in farrowing rate (91.5% × 89.1%) and litter size (12.5 × 11.9 piglets born, respectively for PCAI and CAI sows). Successful passage of the intrauterine catheter in all the inseminations was possible in 86.8% (165/190) of sows initially allocated to PCAI treatment. Difficulty of introducing the catheter in at least one insemination did not affect the reproductive performance of PCAI sows (P > 0.05). Bleeding during insemination did not affect (P > 0.05) the farrowing rate in both treatments, but litter size was reduced in CAI and PCAI sows (P ≤ 0.06). Percentage of spermatozoa present in backflow within 1 h after insemination was greater in CAI than PCAI sows (P < 0.01). More than 85% of primiparous sows can be successfully post‐cervical inseminated with doses containing 1.5 × 10⁹ sperm cells in the absence of the boar during insemination without impairing the reproductive performance.     

N‐Acetyl‐d‐galactosamine prevents soya bean agglutinin‐induced intestinal barrier dysfunction in intestinal porcine epithelial cells

Soya bean agglutinin (SBA) is a glycoprotein and the main anti‐nutritional component in most soya bean feedstuffs. It is mainly a non‐fibre carbohydrate‐based protein and represents about 10% of soya bean‐based anti‐nutritional effects. In this study, we sought to determine the effects of N‐Acetyl‐D‐galactosamine (GalNAc or D‐GalNAc) on the damage induced by SBA on the membrane permeability and tight junction proteins of piglet intestinal epithelium (IPEC‐J2) cells. The IPEC‐J2 cells were pre‐cultured with 0, 0.125 × 10^?4, 0.25 × 10^?4, 0.5 × 10^?4, 1.0 × 10^?4 and 2.0 × 10^?4 mmol/L GalNAc at different time period (1, 2, 4 and 8 hr) before being exposed to 0.5 mg/ml SBA for 24 hr. The results indicate that pre‐incubation with GalNAc mitigates the mechanical barrier injury as reflected by a significant increase in trans‐epithelial electric resistance (TEER) value and a decrease in alkaline phosphatase (ALP) activity in cell culture medium pre‐treated with GalNAc before incubation with SBA as both indicate a reduction in cellular membrane permeability. In addition, mRNA levels of the tight junction proteins occludin and claudin‐3 were lower in the SBA‐treated groups without pre‐treatment with GalNAc. The mRNA expression of occludin was reduced by 17.3% and claudin‐3 by 42% (p < 0.01). Moreover, the corresponding protein expression levels were lowered by 17.8% and 43.5% (p < 0.05) respectively. However, in the GalNAc pre‐treated groups, occludin and claudin‐3 mRNAs were reduced by 1.6% (p > 0.05) and 2.7% (p < 0.01), respectively, while the corresponding proteins were reduced by 4.3% and 7.2% (p < 0.05). In conclusion, GalNAc may prevent the effect of SBA on membrane permeability and tight junction proteins on IPEC‐J2s.     

The Effect of Glycerol Concentrations on the Post‐thaw In Vitro Characteristics of Cryopreserved Sex‐sorted Boar Spermatozoa

The objective of this study was to optimize protocols for the cryopreservation of sex‐sorted boar spermatozoa. In the experiment 1, we evaluated the effects of a standard boar sperm cryopreservation procedure (3% final glycerol concentration) on the in vitro characteristics of sex‐sorted sperm frozen at low sperm concentrations (20 × 10⁶ sperm/ml; S20 group). Non‐sorted spermatozoa frozen at 1000 × 10⁶ (C1000 group) and 20 × 10⁶ (C20 group) sperm/ml were used as the freezing control groups. In experiment 2, the effects of different final glycerol concentrations (0.16%, 0.5%, 1.0%, 2.0% and 3.0%) on post‐thaw quality of the S20 and C20 groups were evaluated. In both experiments, the samples were evaluated prior to freezing (5°C) and at 30, 90 and 150 min after thawing. Experiment 1 indicated that freezing sperm at low concentrations decreased (p < 0.05) the total motility (TM) and progressive motility (PM) at 90 and 150 min after thawing regardless of whether the sperm were sorted or not. However, the sperm membrane integrity was not affected at any evaluation step. Inexperiment 2, significant effects on the TM and PM because of increased glycerol concentrations in the S20 and C20 groups were observed only at 90 and 150 min after thawing. The samples frozen in 3% glycerol showed lower (p < 0.05) TM and PM values when compared to those frozen in the presence of 0.5% and 1% glycerol. In both experiments, non‐sorted control samples displayed higher percentages of spermatozoa with damaged DNA than sorted spermatozoa. In conclusion, the optimization of cryopreservation conditions by decreasing the glycerol concentrations can improve post‐thaw motility of sex‐sorted spermatozoa frozen at low concentrations.     

Glutathione Peroxidase Activity,Plasma Total Antioxidant Capacity,and Urinary F2‐ Isoprostanes as Markers of Oxidative Stress in Anemic Dogs

Background

Oxidative stress plays a role in the pathophysiology of several diseases and has been documented as a contributor to disease in both the human and veterinary literature. One at‐risk cell is the erythrocyte, however, the role of oxidative stress in anemia in dogs has not been widely investigated.

Hypothesis/Objective

Anemic dogs will have an alteration in the activity of glutathione peroxidase (GPx), a decrease in of total antioxidant capacity (TAC), and an increased concentration of urinary 15‐F₂‐isoprostanes (F₂‐IsoP) when compared to healthy dogs.

Animals

40 client‐owned dogs with anemia (PCV <30%) age‐matched to 40 client‐owned healthy control dogs.

Methods

Prospective, cross‐sectional study. Whole blood GPx activity, plasma TAC, and urinary F₂‐isoprostane concentrations were evaluated in each dog and compared between groups.

Results

Anemic dogs had significantly lower GPx activity (43.1 × 10³ +/‐ 1.6 × 10³ U/L) than did dogs in the control group (75.8 × 10³ +/‐ 2.0 × 10³ U/L; P < 0.0001). The GPx activity in dogs with hemolysis (10³ +/‐ 0.8 × 10³ U/L) was not significantly different (P = 0.57) than in dogs with nonhemolytic anemia (43.5 × 10³ +/‐ 1.1 × 10³ U/L). The TAC concentrations (P = 0.15) and urinary F₂‐isoprostanes (P = 0.73) did not significantly differ between groups.

Conclusions and Clinical Importance

Glutathione peroxidase activity was significantly decreased in anemic dogs indicating oxidative stress. Additional studies are warranted to determine if antioxidant supplementation would improve survival and overall outcome as part of a therapeutic regimen for anemic dogs.     

Antiviral Effect of Saccharomyces cerevisiaeβ‐glucan to Swine Influenza Virus by Increased Production of Interferon‐γ and Nitric Oxide

The aim of these experiments was to investigate the potential antiviral effect of Saccharomyces cerevisiaeβ‐glucan on the pneumonia induced by swine influenza virus (SIV). Forty colostrum‐deprived 5‐day‐old piglets were randomly divided into four groups of 10. The 20 pigs in groups 1 and 2 were administered Saccharomyces cerevisiaeβ‐glucan orally (50 mg/day/pig; En‐Bio Technology Co., Ltd) for 3 days before SIV infection and those in groups 3 and 4 were given culture medium/diluent alone. Groups 1 and 3 were inoculated intranasally with 3 ml of tissue culture fluid containing 2 × 10⁶ tissue culture infective doses 50% (TCID₅₀)/ml of SIV and those in groups 2 and 4 were exposed in the same manner to uninfected cell culture supernatant. The microscopic lung lesions induced by SIV infection (group 1 pigs) were significantly more severe than those induced by infection in animals pre‐administered β‐glucan (group 3) (P < 0.05). Significantly more SIV nucleic acid was detected in the lungs of pigs experimentally infected with SIV only (group 1) at 5, 7 and 10 days post‐inoculation (dpi) compared with lungs from pigs pre‐administered β‐glucan and infected with SIV (group 3) (P < 0.05). The concentrations of interferon‐γ (IFN‐γ) and nitric oxide (NO) in bronchoalveolar lavage fluid from pigs pre‐administered β‐glucan and infected with SIV (group 3) were significantly higher than for any other group at 7 and 10 dpi for IFN‐γ, and at 5, 7 and 10 dpi for NO (P < 0.05). Saccharomyces cerevisiaeβ‐glucan reduced the pulmonary lesion score and viral replication rate in SIV‐infected pigs. These findings support the potential application of β‐glucan as prophylactic/treatment agent in influenza virus infection.     

Generation of embryonic stem‐like cells from in vivo‐derived porcine blastocysts at a low concentration of basic fibroblast growth factor

Although basic fibroblast growth factor (bFGF) is an essential factor supporting the maintenance of porcine embryonic stem (ES) cell self‐renewal and pluripotency, its high cost has limited previous studies, and the development of a low‐cost culture system is required. For these systems, in vivo blastocysts were progressively cultured under various conditions consisting of different culture mediums and/or different feeder cell numbers at a low concentration of bFGF. As the results, the sequential culture of in vivo‐derived porcine blastocysts on 5.0 × 10⁵ mouse embryonic fibroblast (MEF) feeder cells in alpha minimum essential medium‐based medium for primary culture, on 2.5 × 10⁵ MEF feeder cells in Mixture medium for the 1st subpassage, and on 2.5 × 10⁵ MEF feeder cells in DMEM/Ham's F10‐based medium for the post‐2nd subpassage could support the establishment and maintenance of porcine ES‐like cells at the low concentration of bFGF. The established porcine ES‐like cells showed ES cell‐specific characteristics such as self‐renewal and pluripotency. We confirmed that porcine ES‐like cells could be generated from in vivo‐derived porcine blastocysts at a low concentration of bFGF.     

Influence of Inseminate Components on Porcine Leucocyte Migration In Vitro and In Vivo after Pre- and Post-Ovulatory Insemination

A post‐breeding migration of leucocytes (PMN) into the uterus is considered to be an important reason for sperm losses. Minimizing such effects may be necessary for successful insemination with low sperm numbers, as required with sex‐sorted spermatozoa. We examined the magnitude of PMN influx 3 h after pre‐ or post‐ovulatory insemination with various combinations of seminal plasma (SP), semen extender Androhep? (AH; Minitüb, Tiefenbach, Germany) and sperm preparations (S). Pre‐ovulatory inseminations with preparations containing 98% AH caused a massive influx of PMN, independent of whether spermatozoa were present (628 ± 189 × 10⁶ leucocytes/uterine horn) or not (580 ± 153 × 10⁶). Post‐ovulatory, 98% AH caused a comparable immigration only in the absence of sperm cells (AH: 569 ± 198 × 10⁶, AH+S: 162 ± 102 × 10⁶). The presence of SP significantly dampened the numbers of recruited uterine leucocytes. The reaction to all inseminates containing 98% SP both with and without spermatozoa, used before ovulation (SP: 14 ± 6 × 10⁶, SP+S: 73 ± 27 × 10⁶) and after ovulation (SP: 60 ± 32 × 10⁶, SP+S: 51 ± 33 × 10⁶) did not differ significantly from controls using phosphate buffered saline (PBS) (pre‐ovulatory: 1 ± 1 × 10⁶, post‐ovulatory: 11 ± 9 × 10⁶). Quantitative in vitro transmigration assays with blood‐derived PMN proved that AH‐induced leucocyte migration into the uterus to be not as a result of direct chemotaxis, because, on account of the chelator citrate, AH significantly inhibited the transmigration towards recombinant human Interleukin‐8 (rhCXCL8) (AH: 14 ± 5% migration rate vs controls: 37 ± 6%, p < 0.05). Supernatants of spermatozoa incubated in PBS for 1, 12 or 24 h showed neither chemoattractive nor chemotaxis‐inhibiting properties. SP at ≥0.1% [v/v] significantly inhibited the in vitro transmigration of PMN. With respect to in vivo migration of neutrophils, the striking difference in the results between semen extender and seminal plasma suggests that adaptation of extender composition is needed to reflect more closely the in vivo regulatory potential of natural seminal plasma.     

Effects of Saccharomyces cerevisiae supplementation on growth performance,plasma metabolites and hormones,and rumen fermentation in Holstein calves during pre‐ and post‐weaning periods

This study aimed to evaluate the effects of supplementing Saccharomyces cerevisiae (SC) during the pre‐ and post‐weaning periods on growth, metabolic and hormonal responses, and rumen fermentation in calves. Three‐week‐old Holstein calves were assigned to either control (n = 12) or SC group (n = 12), the latter of which received 2 × 10⁹ cfu/day of SC. The experiment was conducted over a period of 7 weeks around weaning. Daily gain (DG) in the SC group was higher (p < .05) than that in the control group. In the SC group, plasma glucose, insulin, and growth hormone (GH) concentrations were higher (p < .05) and concentrations of glucagon and insulin‐like growth factor 1 (IGF‐1) tended to be higher (p < .1) than in the control group. Proportion of rumen propionate and concentration of rumen ammonia nitrogen at 10 weeks of age were greater (p < .05) in the SC group than that in the control group. Supplementation of SC around weaning may improve dietary nutrient and energy availability and increase plasma GH and IGF‐1 concentrations. These changes observed in SC‐supplemented calves could be closely related to the improvement of DG.     

Use of the Sperm Quality Analyzer (SQA II‐C) for the Assessment of Dog Sperm Quality

In the present study, an automated system for sperm analysis, the Sperm Quality Analyzer (SQA II‐C), was tested as a potential tool for the assessment of dog sperm quality. In the first experiment the device displayed a good repeatability of measurements for semen of medium and high quality, as evidenced by a low coefficient of variance (CV; 0.08), whereas a high CV (0.46) was obtained for one dog with semen of inferior quality. In the second experiment, seven different sperm concentrations (25–300 ×10⁶/ml), obtained by dilutions in Hepes‐TALP medium were stored for 48 h at room temperature. A concentration dependent increase in sperm motility index (SMI) was shown, reaching a plateau at 150×10⁶ spermatozoa/ml. For all sperm concentrations, the SMI value decreased significantly after 24 h, indicating the importance of sperm motility for SMI values. For sperm concentrations lower than 150×10⁶/ml, highly significant correlations [r=0.80;p<0.05] were established between SMI values on one hand and sperm concentration, and semen parameters expressing the overall semen sample quality on the other hand (experiment 3) while non‐significant or low correlations were found between SMI values and other individual sperm parameters. In experiment 4, significantly high correlations (r=0.97) were found between mean SMI values and post‐thaw motility and progressive motility assessed subjectively. In conclusion, our study indicates that both motility and concentration largely influence SMI values and that the SQA II‐C saturates at 150×10⁶ fresh spermatozoa/ml. In our opinion, the SQA II‐C may be a useful and objective device to assess the post‐thaw motility of dog sperm.     

ORIGINAL ARTICLE: Effects of herbage ingestion upon ileal digestibility of amino acids in heavy Iberian pigs fed on an acorn‐based diet

We conducted two experiments with heavy Iberian pigs to determine the ileal digestibility of amino acids (AA) in acorns and freshly cut herbage, and the effects of adding fresh herbage upon the supply of ileal digestible AA when pigs were fed on holm‐oak acorns. In Experiment 1, carried out in cannulated pigs of 107 kg bodyweight (BW), daily intake of acorns reached 44.9 g DM/kg^0.75 BW. Arg, His and Thr showed the lowest apparent ileal digestibility (AID) values, whereas Met, the branched‐chain AA and Phe had the highest coefficients. The AID of total EAA was 0.716 but only 0.222 for NEAA. Most of the digestive and absorptive processes of acorn protein occurred before the hindgut. Acorn provides (per kg DM) 2.27 g apparent ileal digestible Lys and 22.7 g apparent total digestible AA. Standardized ileal digestibility (SID) values for EAA, NEAA and total AA were 0.924 ± 0.020, 0.784 ± 0.041 and 0.860 ± 0.029. In Experiment 2 fresh herbage was given to six cannulated Iberian pigs of 140 kg either as a single feed (13.7 g DM/kg^0.75 BW) or as a supplement to acorns (28.4 g DM/kg^0.75 BW). When only freshly cut forage was offered the AID of the EAA, NEAA and total AA was close to 0.65 and supplied (per kg DM ingested) 5.61 g AID Lys and 91.7 g digestible AA. Standardized ileal values were 0.744 ± 0.023, 0.912 ± 0.038 and 0.831 ± 0.030 respectively. The addition of fresh forage to the acorns led to a significant decrease in AID of AA in acorn due to digesta transfer to the hindgut: His (p < 0.01), Met (p < 0.001), Phe (p = 0.092), Thr (p < 0.05) and Val (p < 0.05), but Arg, Lys and the branched‐chain AA remained unaffected. The main contribution of herbage to AA nutrition of the grazing Iberian pig relies mainly on increasing the supply of digestible AA for pig tissues.     

Immune‐mediated hemolytic anemia and severe thrombocytopenia in dogs: 12 cases (2001–2008)

Objective – To identify and characterize the syndrome of immune‐mediated hemolytic anemia (IMHA) with concurrent severe thrombocytopenia (≤15.0 × 10⁹ platelets/L; [15.0 × 10³ platelets/μL]), and to evaluate prognostic factors, clinicopathologic findings, complications, treatment, outcome, and survival of dogs with this hematologic disorder. Design – Retrospective, observational study. Setting – Veterinary teaching hospital. Animals – Twelve client‐owned dogs with IMHA and severe thrombocytopenia (≤15.0 × 10⁹ platelets/L; [15.0 × 10³ platelets/μL]), without evidence of overt disseminated intravascular coagulation. Interventions – The following data were recorded and analyzed from the electronic medical record: signalment, history, concurrent diseases, clinical signs at presentation, clinicopathologic data, diagnostic testing, radiographic findings, treatment modalities, length of hospitalization, complications, and clinical outcome. All dogs were treated with immunosuppressive doses of corticosteroids. Measurements and Main Results – Twelve dogs were identified with the diagnosis of IMHA and severe thrombocytopenia; of these, 9 (75%) survived, 3 (25%) were euthanized, and none died. Dogs that survived were significantly younger than nonsurvivors (P=0.03). There were no specific clinical signs or therapies associated with survival. Conclusions – Dogs in this study had a mortality rate similar to reported rates for dogs with either disease alone. Overall, younger dogs were more likely to survive. No association between different treatment modalities and overall survival was identified.     

Effect of Different Levels of Glycerol and Cholesterol‐Loaded Cyclodextrin on Cryosurvival of Ram Spermatozoa

This study was conducted to determine the optimum level of glycerol and cholesterol‐loaded cyclodextrin (CLC) in a Tris‐based diluent for cryopreservation of ram spermatozoa. Ram semen was treated with 0, 1.5, 3 or 4.5 mg CLC/120 × 10⁶ cells in Tris‐based diluents containing 3, 5 or 7% glycerol in a factorial arrangement 3 × 4 and frozen in liquid nitrogen vapour. Sperm motility, viability (eosin–nigrosin staining) and functional membrane integrity (hypo‐osmotic swelling test) were assessed immediately after thawing (0 h) and subsequently after 3 and 6 h at 37°C. There was an interaction between CLC and glycerol on the functional membrane integrity (p < 0.05). In the presence of 3% glycerol, the highest functional membrane integrity (32.2%) was found in the spermatozoa treated with 1.5 mg CLC/120 × 10⁶ sperm. Post‐thaw sperm motility was highest in 1.5 mg CLC immediately after thawing (40.5%) and after 3‐h (30.6%) incubation at 37°C (p < 0.05). Viability of spermatozoa was higher in all CLC treatments than in the untreated samples, and it was highest (33.9%) in the spermatozoa treated with 1.5 mg CLC (p < 0.05). These data indicate that the addition of cholesterol to sperm membranes by 1.5 mg CLC/120 × 10⁶ cells may allow the use of a lower concentration of glycerol (3%), which is sufficient to mitigate the detrimental effects of freezing and thawing.     

Orally administered Chrysin improves post‐thawed sperm quality and fertility of rooster

Chrysin is a bioflavonoid compound found in passion flower, chamomile, propolis and honey at high levels. Post‐thawed sperm quality and fertility of Chrysin‐fed roosters were assessed in this study. Twenty 40‐week‐old male broiler breeders were randomly divided into four groups and fed basal diet supplemented with different levels of Chrysin including 0 (Ch‐0), 25 (Ch‐25), 50 (Ch‐50) or 75 (Ch‐75) mg/day for 12 consecutive weeks. Semen samples were weekly collected from 6th to 9th week of experiment to evaluate some sperm quality parameters including total and progressive motility, plasma membrane integrity and functionality (in fresh and post‐thawed samples) and mitochondrial activity (only in post‐thawed samples). Also, collected semen samples from 10th, 11th and 12th week of experiment were frozen and then artificially inseminated to test fertility rate. According to the results, an improvement in both fresh and post‐thawed sperm quality including total [fresh: 88.00 ± 0.58 and 87.25 ± 0.67 (p < .01); post‐thawed: 51.07 ± 2.05 and 52.72 ± 1.96 (p < .01)] and progressive motility [fresh: 76.00 ± 0.58 and 78.25 ± 0.65 (p < .01); post‐thawed: 40.61 ± 2.01 and 39.88 ± 2.01 (p < .01)], plasma membrane integrity [fresh: 91.60 ± 0.58 and 89.85 ± 0.59 (p < .01); post‐thawed: 56.99 ± 1.86 and 54.39 ± 1.86 (p < .01)] and functionality [fresh: 75.40 ± 0.42 and 77.90 ± 0.96 (p < .01); post‐thawed: 45.69 ± 1.71 and 46.35 ± 1.71 (p < .01)] was noted for both Ch‐50 and Ch‐75, respectively, groups compared to control group. Despite no significant change in mitochondrial activity, fertility rate of post‐thawed spermatozoa was significantly improved in all Chrysin‐fed groups compared to Ch‐0 group. In conclusion, oral Chrysin administration to roosters could ameliorate cryopreservation‐induced impairment of sperm quality and fertility rate.