Differential Expression of IGF Family Members in Heat‐Stressed Embryos Produced In Vitro from OPU‐Derived Oocytes of Nelore (Bos indicus) and Holstein (Bos taurus) Cows

The IGF system is related to embryo quality. We aim to determine the effect of the heat stress on the mRNA expression of IGF1 and IGF2, IGFR1 and IGFR2, IGFBP2 and IGFBP4, and PAPPA in in vitro production (IVP) blastocysts from Nelore and Holstein after ovum pick up (OPU) to better understand the differences between these breeds. Oocytes from four Nelore and seven Holstein were collected in six OPU sessions. Following in vitro maturation and fertilization using six Nelore or Holstein sires, embryos were divided into control (cultured at 39°C) and heat stress (HS; exposed to 41°C for 9 h). Blastocysts were submitted to RNA extraction. The IGF1 expression was higher in blastocysts under HS in both breeds, and the expression of IGFBP2 and IGFBP4 was higher in Holstein blastocysts under HS. The high PAPPA expression and the low expression of IGFBP2 and IGFBP4 are associated with a more efficient degradation of IGFBPs, which results in greater IGF bioavailability in Nelore blastocysts and may contribute to the superior HS tolerance in Nelore, when compared to Holstein.     

Ovine rumen papillae biopsy via oral endoscopy; a rapid and repeatable method for serial sampling

AIMS: To explore and validate the utility of rumen endoscopy for collection of rumen papillae for gene expression measurement.

METHODS: Four adult Coopworth ewes were fasted for either 4 or 24 hours. Animals were sedated, placed in a dorsally recumbent position at 45 degrees with the head upright, and an endoscope inserted via a tube inserted into the mouth. Biopsies of rumen papillae were taken from the ventral surface of the rumen atrium under visual guidance. Two biopsies were collected from one of the animals that had been fasted for 4 hours, and three from one of the animals that had been fasted for 24 hours. Video of the rumen atrium and reticulum was also collected. The animals recovered uneventfully. Biopsies were subsequently used for extraction and sequencing of mRNA.

RESULTS: The ventral surface of the rumen atrium was accessible after 4 hours off pasture, but a larger region was accessible after 24 hours of fasting. Sedation allowed access for endoscope use for around 5 to 10 minutes after which increased saliva flow was noted. Rumen papillae biopsies were easily collected, with samples from a variety of sites collected in the ～10 minute time window. High quality RNA was obtained for stranded mRNA sequencing. Of the resulting reads, 69–70% mapped uniquely to version 3.1 of the ovine genome, and 48–49% to a known gene. The rumen mRNA profiles were consistent with a previously reported study.

CONCLUSIONS: This method for obtaining rumenal tissue was found to be rapid and resulted in no apparent short or long term effects on the animal. High quality RNA was successfully extracted and amplified from the rumen papillae biopsies, indicating that this technique could be used for future gene expression studies. The use of rumen endoscopy could be extended to collection of a variety of rumen and reticulum anatomical measurements and deposition and retrieval of small sensors from the rumen. Rumen endoscopy offers an attractive and cost effective approach to repeated rumen biopsies compared with serial slaughter or use of cannulated animals.     

Inoculation of Sarcocystis neurona merozoites into the central nervous system of horses

Equine protozoal myeloencephalitis (EPM) is a neurologic syndrome in horses from the Americas and is usually caused by infection with the apicomplexan parasite, Sarcocystis neurona. A horse model of EPM is needed to test the efficacy of chemotherapeutic agents and potential vaccines. Five horses that were negative for antibodies to S. neurona in their serum and cerebrospinal fluid (CSF) were injected in the subarachnoid space with living merozoites of the SN2 isolate of S. neurona. None of the horses developed clinical disease or died over a 132-day observation period. All five horses developed antibodies to S. neurona in their CSF and serum 3-4 weeks after injection. Two of the horses were examined at necropsy and no parasite induced lesions were observed in their tissues and no parasites were recovered from portions of their spinal cords inoculated on to cell cultures. Results of this study demonstrate that merozoites of the SN2 isolate of S. neurona will induce seroconversion but not clinical disease when inoculated directly into the CSF of nonimmune horses.     

Plasma Antimullerian Hormone as a Predictor of Ovarian Antral Follicular Population in Bos indicus (Nelore) and Bos taurus (Holstein) Heifers

In Bos taurus cattle, antimullerian hormone (AMH) has been demonstrated to have a high degree of correlation with ovarian antral follicle count and the number of healthy follicles and oocytes. To document the correlation between the plasma concentration of AMH and follicular number in Bos indicus and Bos taurus heifers, Nelore (Bos indicus, n = 16) and Holstein heifers (Bos taurus, n = 16) had their ovarian follicular waves synchronized. After synchronization, ovarian antral follicular population (AFP) was evaluated three times at 60‐day (d) intervals (T‐120 d, 120 days before plasma AMH determination; T‐60 d, 60 days before; and T0, at the time of plasma AMH determination). The plasma AMH concentration was positively correlated with the number of ovarian follicles on the day of the follicular wave emergence in Bos indicus (Nelore) and Bos taurus (Holstein) heifers at each evaluation time (p < 0.05). The AFP was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.05). Similarly, the AMH concentration was higher in Bos indicus (Nelore) than in Bos taurus (Holstein) heifers (p < 0.0001). When heifers were classified as to present high or low AFP according to the mean of the AFP within each genetic group, high‐AFP heifers presented a greater (p < 0.0001) AMH concentration than low‐AFP heifers, regardless of the genetic group. In conclusion, the AFP is positively correlated with plasma AMH concentration in both Bos indicus (Nelore) and Bos taurus (Holstein) heifers. Furthermore, Bos indicus (Nelore) heifers presented both greater plasma AMH concentrations and AFP than Bos taurus (Holstein) heifers.     

Comparison of lameness outcomes in horses with acute or chronic digital lameness that underwent magnetic resonance imaging

ABSTRACT

Aims: To compare the outcome, in terms of lameness score or return to athletic function, of horses with acute vs. chronic digital lameness that underwent magnetic resonance imaging (MRI) of the distal limb and to compare the proportion of horses that received intra-articular therapy of the distal interphalangeal (DIP) joint and pattern of diagnostic analgesia in these groups.

Methods: This is a retrospective study of horses (n?=?95) with acute (≤12 weeks; n?=?46) or chronic (>12 weeks; n?=?49) digital lameness that underwent MRI of the distal limb from 2009–2016, at two equine referral centres in the USA. Criteria for inclusion in the study were that a majority of lameness localised distal to the fetlock, and that lameness assessments for ≥12 months following MRI could be obtained from the medical record or the owner could be interviewed regarding their horse's athletic function. Outcome was characterised by an improvement score where 2?=?return to work at a previous or higher level or lameness improved by one grade or more, 1?=?return to work at a lower level or lameness improved by less than one grade, and 0?=?did not return to work or lameness grade worsened. Whether horses had received intra-articular therapy of the DIP joint and the pattern of diagnostic analgesia prior to MRI was also obtained from medical records or by interviewing the owner.

Results: There was a difference (p?=?0.004) in the proportion of horses assigned to improvement scores of 0, 1 and 2 between horses with acute or chronic lameness. There was no evidence of a difference in the likelihood of having received intra-articular therapy of the DIP joint prior to MRI between horses with chronic or acute lameness (p = 0.085). Similarly, there was no evidence of a difference in the pattern of diagnostic analgesia prior to MRI between the two groups (p = 0.94). Eighty-two percent of owners of horses with acute and 62% of those with horses with chronic lameness had a positive opinion of the utility of MRI as a diagnostic modality.

Conclusion: In a population of horses with digital lameness undergoing MRI, a difference in the outcome, in terms of lameness score or return to athletic function was identified between horses with acute lameness compared to those with chronic lameness.

Clinical relevance: Horses with digital lameness that undergo MRI when the lameness is acute may have an improved prognosis due to accurate diagnosis and earlier application of appropriate therapy.     

Wie viele Nährstoffe werden mit dem Mulchen der Fahrgassen in den Baumstreifen einer Obstanlage verfrachtet?

The objective of this PhD project was to investigate the nutrient translocation associated with the mulch from the grass alley to the tree strip. Therefore, a historical, approved 40-year fertiliser trial with apple cv. ‘Gloster’ on M9 rootstock at a spacing of 4 m?×?1.5?m was employed at Klein-Altendorf Research Centre, University of Bonn, Germany, which comprised mineral nutrition (1?t/ha??CaO? $ + $ ?NPK), an organic plot (10?t??FM/ha pig manure) and an unfertilised plot as control. Grass alleys were mowed 5?× in 1995 and 3?× in 1996 as commonly practised in fruit orchards; foil was spread on the tree strips to capture the biomass transferred with the mulching. Overall, ca.??60% of the grass mulch dry matter (mowed at 2?cm height) remained in the grass alleyway, while ca.??40% was translocated to the tree strips, which amounted to 550?kg??DM (Dry Matter) in the unfertilised plot and 800?kg??DM/ha/year in both fertilised plots with the five mulches in the wet year 1995. Grass mulch contained 2–3%??N and 2.2–2.9%??K in the unfertilised control plot or 2.1–3.9%??N and 2.7–4.1%??K on per leaf dry matter in both the inorganic NPK and organically fertilised plot; the nitrogen content of the grass mulch increased during the growing season in the wet year 1995, but remained low and constant at 2.5%??N in the dry year 1996. By contrast, calcium was decreased from 0.5–0.7%??Ca in the unfertilised plot to 0.3–0.6%??Ca in the organic plot. Overall, 25–50% of the grass mulch was translocated from the grass alleys (ca. 4,000 m²) into the tree strips, representing an organic fertilisation of 10–25?kg??N, 2–4?kg??P, 15–25?kg??K, 3–5?kg??Ca and 1–2?kg??Mg per hectare and year. This would theoretically be equivalent to ca 50% supply of??N,??P and??K and??10% of calcium as required nutrients by fruit trees depending on rate of nutrient availability. Its subsequent partial availability reported of 54%??N within??90 days after mulching and large nutrient content are discussed with respect to the low nutrient content in leaves dropped from the fruit trees in the autumn, where the nutrients had been previously translocated into the woody perennial parts of the tree. This nutrient cycling should be integrated in environmental-friendly sustainable fertilisation strategies for fruit orchards.