Suberoylanilide hydroxamic acid,a novel histone deacetylase inhibitor,improves the development and acetylation level of miniature porcine handmade cloning embryos

Histone deacetylase inhibitors (HDACis) can change the histone acetylation and significantly enhance the developmental competence of the pre‐implantation SCNT embryo. To select a proper histone deacetylase inhibitor to improve the success rate and potentially developmental ability of handmade cloning (HMC) embryos of miniature porcine, we compared the effect of two histone deacetylase inhibitors (SAHA vs. VPA) on HMC embryo development, their histone acetylation level and the expression level of relevant genes. The blastocyst rate and number of blastocyst cells of HMC embryos treated with SAHA (SAHA‐HMC) or VPA (VPA‐HMC) were significantly higher than those of control (Control‐HMC), respectively, but there were no significant difference between SAHA‐HMC and VPA‐HMC groups. In addition, the acetylation level (AcH4K8) of Control‐HMC and VPA‐HMC embryos at the blastocyst stage, respectively, was significantly lower than that of in vitro fertilized (IVF) and SAHA‐HMC embryos. However, the acetylation H4K8 of the blastocysts had no significant difference between SAHA‐HMC and the IVF groups. The SAHA‐HMC blastocysts indicated comparative expression levels of Oct4 and HDAC1 (histone deacetyltransferase gene) with those of IVF blastocysts. In contrast, the expression levels of Oct4 were lower and those of HDAC1 were higher in the VPA‐HMC and Control‐HMC blastocysts, respectively, compared to those of the IVF blastocysts. Our results demonstrated that the HMC embryos treated by SAHA could promote the pre‐implantation development and increase the levels of histone H4K8 acetylation and the expression of the OCT4 gene, yet decrease the expression of the HDAC1 gene to the comparable level of the IVF embryos. Our results proved that SAHA may be a better histone deacetylase inhibitor for porcine HMC compared to VPA, and furthermore, it may indicate that SAHA can effectively correct the abnormal histone acetylation during the HMC embryo development and subsequently improve the full‐term developmental potential of the HMC embryos after embryo transplantation.     

European Eel Sperm Diluent for Short‐term Storage

The sperm of European eel shows a high density and the time of spermatozoa motility is very short after activation with sea water. These characteristics make difficult the sperm handling and its quality assessment. Several diluents were previously described for the Japanese eel obtaining over 3 weeks’ conservation times under refrigeration, but they rendered bad results in the European species. In the present study, several diluents were developed taking as basis the P1 medium, and using different dilution ratios (1 : 50, 1 : 100) and two pH (6.5, 8.5). The effect of the addition of bovine serum albumin (BSA, 2% w/v) was also evaluated. At 24 h, undiluted samples already showed significant lower motility and viability than sperm samples diluted in the different media. The results for diluents with pH 6.5 and 8.5 were different. Spermatozoa diluted in media at pH 6.5 cannot be activated at 24 h, while samples diluted in the diluents with pH 8.5 and added with BSA did not show significant differences with respect to the fresh sperm motility until 48 h. The viability (percentage of alive cells) did not show differences until 1 week, independent of the dilution ratio. After 1 week, the motility was approximately 30% in the media containing BSA, which presented no differences for head size of the spermatozoa (perimeter and area) until 72 h and 1 week, respectively. In conclusion, the combination of one medium having similar physico‐chemical characteristics to the seminal plasma, including pH 8.5, and supplemented with BSA can be used in different dilution ratios for the sperm’s short‐term storage, preserving its motility capacity.     

Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

Cytopathological and histopathological diagnosis of canine splenic disorders

Objectives To determine (1) the common types of canine splenic disorders, and the breeds affected, that are diagnosed by cytopathological and histopathological examination in Sydney, Australia and (2) the accuracy of cytopathological examination compared with histopathological examination for the diagnosis of canine splenic disorders. Design 69 cytopathological and 51 histopathological diagnoses of canine splenic disorders presented to the Veterinary Pathology Diagnostic Services, The University of Sydney during 2006 and 2007 were tabulated and analysed; 17 cases examined both cytopathologically and histopathologically during 2001–07 were also analysed. Results The most common cytopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (29%), followed by no abnormalities detectable (28%), malignant neoplasms (20%), equivocal diagnoses (20%) and inflammatory disorders (3%). The most common breeds were Kelpie crosses and mixed breeds. The most common histopathological diagnoses were benign disorders of growth, vascular disturbances and necrosis (49%), followed by malignant neoplasms (43%) and inflammatory disorders (8%). The most common breeds were German Shepherd Dogs, Boxers and Maltese Terriers. Cytopathological and histopathological diagnoses were in complete agreement in 59% of cases, partial agreement in 29% and disagreement in 12%. Conclusion Benign disorders of growth, vascular disturbances and necrosis were the most commonly diagnosed canine splenic disorders, both cytopathologically and histopathologically. Kelpie crosses presented most frequently for cytopathological examination. German Shepherd Dogs were the most common breed diagnosed histopathologically with haemangiosarcoma. Although cytopathological and histopathological splenic examinations are complementary for diagnosis, this study has shown a high correlation for complete and partial agreement between the two.     

A survey of northern Victorian dairy farmers to investigate dairy calf management: colostrum feeding and management

Objectives

To describe colostrum management practices carried out in northern Victorian dairy herds and to identify weaknesses in these areas that may affect calf health and welfare by comparing the results with the current industry recommendations

Methods

A questionnaire to obtain information about colostrum management and calf‐rearing practices was sent to commercial dairy farming clients of Rochester Veterinary Practice between June and September 2013. The questionnaire consisted of a general herd overview and colostrum harvesting practices.

Results

The response rate was 39% (58/150). Many dairy producers were not meeting the current industry recommendations in the following areas: (1) time of removal calf from the dam, (2) relying on calf suckling colostrum from the dam to achieve adequate passive transfer, (3) failing to supplement calves with colostrum, (4) feeding inadequate volumes of colostrum, (5) delayed colostrum harvesting, (6) pooling of colostrum, (7) failing to objectively assess colostrum quality or relying on visual assessment and (8) storing colostrum for a prolonged periods of time at ambient temperatures.

Conclusion

The results from this survey highlight the need for greater awareness of industry standards for colostrum management and feeding hygiene.

     

Scrotal circumference,bodyweight and semen characteristics in growing dairy-breed natural-service bulls in Tasmania,Australia

Aims: To provide herd managers with a set of decision rules allowing them to predict the likelihood that a juvenile bull is ready for Bull Breeding Soundness Evaluation (BBSE), or breeding, if bodyweight and scrotal circumference are known.

Methods: This was a longitudinal study following two groups of young pasture-fed Holstein and Jersey bulls from northwest Tasmania, Australia. Individual scrotal circumference, bodyweight and semen characteristics were recorded at 6–8 weekly intervals, from 6–18 months of age. Classification and regression tree analyses were used to predict the probability that a bull had ≥70% normal sperm morphology based on scrotal circumference and bodyweight measurements.

Results: Overall 1,661 scrotal circumference and bodyweight measurements were obtained, and 518 semen samples from 356 bulls were assessed for sperm morphology, from 16 examination sessions that took place between 29 May 2015 and 17 August 2016. Classification and regression tree analyses generated a decision tree for Holstein bulls with four node endpoints, and for Jersey bulls with three node endpoints. Diagnostic test performance showed that for Holstein bulls, using the node endpoints of scrotal circumference ≥27?cm and bodyweight ≥349?kg, 98% had ≥70% normal sperm (positive likelihood ratio 10.4; 95% CI?=?2.7–41), and using the node endpoints of scrotal circumference ≥27?cm and bodyweight between 282–349?kg, 89% had ≥70% normal sperm (positive likelihood ratio 1.6; 95% CI?=?0.9–2.6). For Jersey bulls, using the node endpoints of bodyweight ≥259?kg and scrotal circumference ≥29?cm, 88% had ≥70% normal sperm (positive likelihood ratio 3.4; 95% CI?=?1.6–7.0).

Conclusions: This study provides a set of relatively simple decision rules based on bodyweight and scrotal circumference measurements that allows herd managers to assess the likelihood that juvenile bulls are ready for BBSE or breeding.

Abbreviations: BBSE: Bull breeding soundness evaluation; BRT: Boosted regression tree     

Soil and agronomic factors associated with cadmium accumulations in kidneys of grazing sheep

SUMMARY Mean concentration of cadmium (Cd) in kidneys of hogget sheep from 67 flocks grazing in the Agricultural Region of Western Australia was tested for association with soil, pastoral, climatic and nutritional factors. Hoggets grazing pastures on acidic soils and soils with a sandy-textured surface had higher Cd concentrations in kidneys than hoggets grazing pastures on more alkaline soils or soils with a clay-textured surface. Application of more than 100 kg of phosphatic fertiliser during the past 3 years to loamy soils was also associated with greater Cd concentration in kidneys of the grazing animals.     

Pilot study--parenteral treatment of recently acquired subclinical mastitis during lactation

OBJECTIVE: To conduct a pilot study to investigate the effect of parenteral antibiotic treatment with penethamate hydriodide on recently acquired sub-clinical mastitis. DESIGN: Sixty-nine cows from five Western Victorian dairy farms were enrolled in a pilot trial. Cows that had an Individual Cow Cell Count (ICCC) of greater than 250,000 cells per mL in their milk at their first or second herd test were enrolled in the trial if they were in their first lactation, or if they had a history of ICCCs below 250,000 cells per mL at all herd tests during the previous lactation. These criteria were used in an effort to exclude cows with chronic subclinical mastitis and include cows with recently acquired subclinical mastitis. PROCEDURE: Cows were divided into control and treatment groups on the basis of having an odd or even ear tag number. The treatment group was treated with penethamate hydriodide shortly after the first or second herd test; the control group was left untreated. The ICCCs of subsequent herd tests were recorded for the remainder of lactation. RESULTS: In 58% of the treatment group, and 25% of the control group, all subsequent ICCCs for that lactation were less than 250,000 cells per mL (p < 0.05). If only first and second lactation cows were considered, 72% of the treatment group, and 11% of the control group, had subsequent ICCCs less than 250,000 cells per mL (p < 0.05). CONCLUSION: An ICCC of greater than 250,000 cells per mL is a good predictor for more high ICCCs at subsequent herd tests. This effect was significantly altered in our trial by treatment with penethamate hydriodide at or near the first or second herd test. Whilst in general treatment of high ICCC cows during lactation is considered to be unrewarding, this pilot study suggests that treatment of cows and heifers appropriately selected on the basis of a previous history of low ICCCs can lead to sustained reductions in cell counts. These results support the case for further study into this area.     

Excitation Gap in the Normal State of Underdoped Bi2Sr2CaCu2O8+delta

Angle-resolved photoemission experiments reveal evidence of an energy gap in the normal state excitation spectrum of the cuprate superconductor Bi2Sr2CaCu2O8+delta. This gap exists only in underdoped samples and closes around the doping level at which the superconducting transition temperature Tc is a maximum. The momentum dependence and magnitude of the gap closely resemble those of the dx2-y2 gap observed in the superconducting state. This observation is consistent with results from several other experimental techniques, which also indicate the presence of a gap in the normal state. Some possible theoretical explanations for this effect are reviewed.