Effects of IGF‐1 on In Vitro Culture of Bovine Preantral Follicles are Dose‐Dependent

This study aimed at assessing the effect of different concentrations of the growth factor similar to insulin 1 (IGF‐1) in the development, survival and ultrastructure of the bovine preantral follicles cultured in situ. Fragments of bovine ovarian cortical tissue were cultured during 1 and 7 days in 1 ml of α‐MEM⁺, supplemented with different concentrations of human recombinant IGF‐1 (0, 30, 70 and 100 ng/ml), in an incubator at 37°C and 5% of CO₂ in 24‐well plates with total replacement of the medium every 2 days. Non‐cultured ovarian fragments (control) and ovarian fragments cultured during 1 and 7 days were processed for classic histology, mechanical isolation and electron transmission microscopy (ETM). Parameters such as normality, viability, activation, development, diameter and ultrastructure were evaluated. All statistical analyses were carried out using sas Version 9.2. The results showed that the percentage of follicles morphologically normal in the IGF‐1 30 ng/ml treatment was similar to the fresh control (p > 0.05) both on the day 1 and on the day 7 of in vitro culture. In the viability analysis, the cultured treatments maintained the percentage of viable follicles during the entire culture period (p > 0.05). After 7 days of culture, the IGF‐1 30 ng/ml treatment showed higher percentages of developing follicles (48.33%) than those of the fresh control (22.22%) and the cultured treatments (p < 0.05). Also, after 7 days of culture, IGF‐1 30 ng/ml presented a higher follicular diameter when compared to the control and other concentrations of IGF‐1 tested. Ultrastructurally, the non‐cultured control and IGF‐1 30 ng/ml, after 7 days of culture, showed conserved oocytes, nuclei and organelles. Hence, it is concluded that IGF‐1 30 ng/ml was the most efficient concentration for the development of bovine preantral follicles cultured in vitro.     

Facial eczema management protocols used on dairy farms in the North Island of New Zealand and associated concentrations of zinc in serum

AIMS: To describe and evaluate the current practices used to manage and prevent facial eczema (FE) in North Island dairy herds, and determine the within-herd prevalence of cows with elevated activities of gamma glutamyl transferase (GGT), and with concentrations of Zn in serum <18?μmol/L.

METHODS: Between January and May 2014, 105 herd managers from throughout the North Island of New Zealand were invited to participate in the study when regional spore counts for Pithomyces chartarum started to rise towards 30,000 spores/g pasture. Managers selected 10 representative cattle that were weighed and blood-sampled by the herd veterinarian. Blood samples were analysed for concentrations of Zn in serum and GGT activity. Pasture samples were also collected and submitted for spore count estimation. Finally a survey of farm management practices relating to prevention of FE was completed by the herd manager. A mixed-effects logistic regression model was used to determine associations between herd-level and cow-level explanatory variables and the probability of a cow having a concentration of Zn in serum <18?µmol/L.

RESULTS: Of the 1,071 cows tested, 79 (7.3 (95% CI=5.8–9.0)%) had GGT activity in serum >300?IU/L, and 35/106 (33 (95% CI=24.2–42.8)%) herds had ≥1 of the 10 cows sampled with GGT activity >300?IU/L. Of the 911 cows that were being treated with Zn, concentrations of Zn were between 18–35?μmol/L in 398 (43.6 (95% CI=40.4–46.9)%) cows, were >35?μmol/L in 32 (3.5 (95% CI=2.4–4.1)%) cows, and <18?μmol/L in 479 (52.6 (95% CI=49.3–55.9)%) cows. After adjusting for the confounding effect of region, the odds of a cow having concentrations of Zn in serum <18?μmol/L were 5.5 (95% CI=1.1–29) times greater for cows supplemented with zinc in water compared with those supplemented by drenching. Of the 105 herd managers, 103 (98%) stated that they had access to regional spore count data, but only 35/105 (33%) reported that they measured spore counts on their own farm. Overall, 98/105 (93%) managers reported that they had some form of FE management programme in place. Fungicides were used on their own or in combination with zinc treatments in 10 herds, ZnSO₄ in water troughs was used in 68 herds, oral drenching with ZnO in nine herds, and ZnO supplied in-feed in 26 herds. Estimated daily dose rates of zinc were less than that required to treat a 400?kg cow on 42/68 farms that administered ZnSO₄ in the water or ZnO as a drench.

CONCLUSION AND CLINICAL RELEVANCE: This study has shown that FE management on dairy farms in the North Island of New Zealand could be substantially improved. It is likely that improved FE management would occur if herd managers were provided with more feedback on the success (or otherwise) of their FE management programmes.     

Endometrial nitric oxide synthase activity in mares susceptible or resistant to persistent breeding‐induced endometritis and the effect of a specific iNOS inhibitor in vitro

Emerging research suggests that the nitric oxide system may play a role in persistent breeding‐induced endometritis (PBIE) in the mare. Differences in uterine nitric oxide (NO) levels between mares susceptible or resistant to PBIE and a dose‐dependent inhibitory effect of NO on uterine contractility have been demonstrated. The objectives of this study were to investigate the difference in total nitric oxide synthase (NOS) activity of the endometrium between susceptible and resistant mares and the effect of a specific inducible nitric oxide synthase (iNOS) inhibitor on the endometrial NOS activity in vitro. Six susceptible and six resistant mares were selected based on preset criteria and the results of an intrauterine challenge with killed spermatozoa during oestrus. Endometrial biopsy samples were collected 24 hr post‐challenge and cultured at 37°C for 24 hr in L‐arginine supplemented minimum essential medium with or without a specific iNOS inhibitor (1,400 W dihydrochloride, 1 mM). The medium and the cultured endometrial tissue were collected after 24 hr of culture and assayed for NO and total protein, respectively. Total NO content of the medium, normalized to endometrial tissue wet weight or total protein, was used as a measure of endometrial NOS activity. Non‐parametric tests were applied for statistical analysis. Susceptible mares had significantly greater endometrial NOS activity than resistant mares. The iNOS inhibitor treatment significantly reduced NOS activity in endometrial samples derived from susceptible and resistant mares. These findings provide a basis for in vivo testing of specific iNOS inhibitors as preventative or therapeutic options for PBIE in mares.     

Landscape of somatic retrotransposition in human cancers

Transposable elements (TEs) are abundant in the human genome, and some are capable of generating new insertions through RNA intermediates. In cancer, the disruption of cellular mechanisms that normally suppress TE activity may facilitate mutagenic retrotranspositions. We performed single-nucleotide resolution analysis of TE insertions in 43 high-coverage whole-genome sequencing data sets from five cancer types. We identified 194 high-confidence somatic TE insertions, as well as thousands of polymorphic TE insertions in matched normal genomes. Somatic insertions were present in epithelial tumors but not in blood or brain cancers. Somatic L1 insertions tend to occur in genes that are commonly mutated in cancer, disrupt the expression of the target genes, and are biased toward regions of cancer-specific DNA hypomethylation, highlighting their potential impact in tumorigenesis.     

Associations between management practices and colostrum quality on New Zealand dairy farms

Abstract

AIMS: To describe colostrum quality in spring-calving dairy herds in New Zealand, in terms of Brix, pH and total and coliform bacterial counts and to investigate associations between farm management practices and these measures of colostrum quality.

METHODS: In June 2015, commercial dairy farms (n=105), located in North and South Islands of New Zealand, were visited shortly after the first cows had calved, and when approximately 50% and 80% of the herd had calved (early, middle and late visits). One litre of pooled colostrum that was being fed to newborn calves was collected at each visit and used to determine Brix, pH, total bacterial and coliform counts. A survey of calf management practices was conducted with the herd manager or calf rearer after the final visit.

RESULTS: Of 298 pooled colostrum samples tested 29/298 (9.7%) had Brix >22%. Brix was higher on farms where calves were picked up twice daily compared with once daily (18.2 (95% CI=16.5–19.9)% vs. 15.9% (95% CI=15.2–16.6)%; p=0.012), and was lower where first milking colostrum was combined with colostrum obtained at later milkings (15.0 (95% CI=13.9–16.1)%) compared with where it was not (16.9 (95% CI=16.3–17.6)%; p=0.002). Vaccination of all cows against calf diarrhoeal pathogens was associated with increased Brix compared with no vaccination (18.1 (95% CI=16.6–19.6)% vs. 16.3 (95% CI=15.6–17.0)%; p=0.033). Mean pH of samples tested decreased from 5.97 (95% CI=5.84–6.09) to 5.58 (95% CI=5.45–5.71) for early and late-season visits, respectively (p<0.001). Of 268 samples tested, 23 (8.6%) had bacterial counts below the recommended threshold of 1.00×10⁵ cfu/mL. Mean bacterial counts increased from 2.75 (95% CI=1.80–3.70)×10⁸ to 4.99 (95% CI=3.95–6.03)×10⁸ cfu/mL for early and late-season visits, respectively (p<0.001). Of 259 samples tested, 23 (8.9%) had coliform counts below the recommended threshold of 1.00×10⁴ cfu/mL.

CONCLUSIONS AND CLINICAL RELEVANCE: On a large majority of dairy farms included in this study the pooled colostrum fed to newborn calves had sub-optimal Brix and excessive bacterial counts. Farm-level risk factors such as twice daily pick up of calves from the paddocks, herd vaccination and feeding pooled colostrum that did not combine colostrum obtained at later milkings with first milking colostrum were associated with colostrum quality measures.     

Calf and colostrum management practices on New Zealand dairy farms and their associations with concentrations of total protein in calf serum

AIMS: To gather information on management practices and farmer attitudes to management of cows and calves during the immediate post-partum period on dairy farms in New Zealand, and to assess these practices for associations with concentrations of total protein (TP) in serum of calves 1–8 days-old.

METHOD: Between July and September 2015 blood samples were collected from calves aged between 24 hours and 7 days, from dairy farms (n=105) in nine areas in New Zealand, on three occasions throughout the calving period. Concentrations of TP were determined in all serum samples. At each visit technicians collected 1?L of the pooled colostrum that was intended for feeding to newborn calves that day. These samples were assessed for Brix, coliform and total bacterial counts. After the last sampling visit, the calf rearer or farm manager were asked to complete a questionnaire describing calf and colostrum management practices on the farm. Potential farm-level variables associated with concentrations of TP in serum of sampled calves were identified using univariable and multivariable linear mixed models.

RESULTS: Mean concentration of TP in serum of calves across all farms was 59.8 (95% CI=59.4–60.2) g/L, and was associated with region and herd size in the final multivariable model. Concentrations of TP were lower in calves from farms in Otago (56.2 (95% CI=53.4–58.9) g/L) and Southland (56.9 (95% CI=54.1–59.7) g/L) compared to calves on farms in the Far North (62.6 (95% CI=59.8–65.3) g/L), and were lower in calves from farms with a herd size >600 (58.3 (95% CI=56.7–59.8) g/L) than ≤600 (61.3 (95% CI=60.1–62.5) g/L) cows. After accounting for fixed effects, farm accounted for only 8.4% of the unexplained variation. There was no association between any of the measures of colostrum quality and concentrations of TP in serum (p>0.2).

CONCLUSION AND CLINICAL RELEVANCE: Very few herd-level variables were associated with concentrations of TP in serum. Risk factors that have been shown to be of importance in previous studies outside New Zealand were not identified as important in the current study. It is possible that, in the situation where calves are kept at pasture with their dams for prolonged periods, variables which influence how well a cow can feed its calf in the first 12–24 hours have a larger influence on concentrations of TP in serum than the collection and management of calves once they reach the rearing shed.     

Influence of preservation methods on the quality of colostrum sourced from New Zealand dairy farms

Abstract

AIMS: To assess the effect of two temperatures (ambient temperature and 4°C), three preservation methods (no preservative, yoghurt and potassium sorbate), and two periods of storage (3 and 7 days) on Brix and total bacterial and coliform counts of colostrum collected from New Zealand dairy farms.

METHODS: One litre of colostrum destined to be fed to newborn calves was collected from 55 New Zealand dairy farms in the spring of 2015. Six aliquots of 150 mL were obtained from each colostrum sample, with two aliquots left untreated, two treated with potassium sorbate and two with yoghurt, and one of each pair of aliquots stored at ambient temperature and the other at 4°C. All samples were tested for Brix, total bacterial counts and coliform counts before treatment (Day 0), and after 3 and 7 days of storage. The effect of preservation method and storage temperature on the change in Brix, bacterial and coliform counts after 3 or 7 days of storage was analysed using multivariable random effects models.

RESULTS: For all outcome variables there was a temperature by preservation interaction. For aliquots preserved with potassium sorbate, changes in Brix and bacterial counts did not differ between aliquots stored at ambient temperature or 4°C, but for aliquots preserved with yoghurt or no preservative the decrease in Brix and increase in bacterial counts was greater for aliquots stored at ambient temperature than 4°C (p<0.001). For aliquots preserved with potassium sorbate, coliform counts decreased at both temperatures, but for aliquots preserved with yoghurt or no preservative coliform counts increased for aliquots stored at 4°C, but generally decreased at ambient temperatures (p<0.001). There was also an interaction between duration of storage and temperature for bacterial counts (p<0.001). The difference in the increase in bacterial counts between aliquots stored at 4°C and ambient temperature after 3 days was greater than between aliquots stored at 4°C and ambient temperature after 7 days.

CONCLUSIONS AND CLINICAL RELEVANCE: Use of potassium sorbate to preserve colostrum for 3 or 7 days resulted in little or no reduction in Brix and a lower increase in total bacterial counts than colostrum stored without preservative or with yoghurt added. Colostrum quality was not affected by storage temperature for samples preserved with potassium sorbate, but storage at 4°C resulted in better quality colostrum than storage at ambient temperatures for colostrum with no preservative or yoghurt added.     

Development of One vs Multiple Ovulatory Follicles and Associated Systemic Hormone Concentrations in Mares

Ablation of follicles ≥  6 mm in diameter and treatment with PGF2α 10 days after ovulation were used to induce the development of ovulatory waves. Comparisons were made between induced waves with one (33 waves, 72%) and multiple (13 waves, 28%) ovulatory follicles. Diameter deviation was defined as the separation of follicles into dominant and subordinate categories. Multiple ovulatory follicles were preceded by more (p < 0.001) follicles ≥ 20 mm at the beginning of deviation, higher LH preceding deviation (approached significance, p < 0.08), lower (p < 0.05) concentrations of FSH on the day of deviation and thereafter, and higher (p < 0.0003) oestradiol by 2 days after deviation. During the peri-ovulatory period, systemic hormone concentrations for waves with multiple ovulations involved higher oestradiol before ovulation (approached significance, p < 0.07), lower FSH (p < 0.04) before and after ovulation, and both higher progesterone (p < 0.05) and lower LH (p < 0.05) beginning the day after ovulation. Results indicated that by the beginning of deviation there were more follicles ≥  20 mm and subsequently greater oestradiol production in waves that led to the development of multiple ovulatory follicles, and during the peri-ovulatory period differences between one and multiple ovulations were consistent with the negative effects of the ovarian hormones on the gonadotropins.     

Origin of storm footprints on the sea seen by synthetic aperture radar

Spaceborne synthetic aperture radar can detect storm footprints on the sea. Coastal weather radar from Cape Hatteras provides evidence that the echo-free hole at the footprint core is the result of wave damping by rain. The increased radar cross section of the sea surrounding the echo-free hole results from the divergence of the precipitation-forced downdraft impacting the sea. The footprint boundary is the gust front; its oriention is aligned with the direction of the winds aloft, which are transported down with the downdraft, and its length implies downdraft impact 1 hour earlier at a quasi-stationary impact spot. The steady, localized nature of the storm remains a mystery.