Prevalence of the AMHR2 mutation in Miniature Schnauzers and genetic investigation of a Belgian Malinois with persistent Müllerian duct syndrome

Persistent Müllerian duct syndrome (PMDS) is a sex‐limited disorder in which males develop portions of the female reproductive tract. Important consequences of PMDS are cryptorchidism and its sequelae of infertility and increased risk of testicular cancer. Anti‐Müllerian hormone (AMH) and its receptor (AMHR2) induce the regression of the Müllerian ducts in male embryos. In Miniature Schnauzer dogs, the genetic basis has been identified as an autosomal recessive nonsense mutation in AMHR2, but the allele frequency of the mutation is unknown. Thus, the primary objective of this study was to estimate the prevalence of the AMHR2 mutation in North American Miniature Schnauzers, in order to ascertain the value of genetic testing in this breed. An additional objective was to determine whether mutations in AMH or AMHR2 were responsible for PMDS in a Belgian Malinois; this would aid development of a genetic test for the Belgian Malinois breed. Genomic DNA from 216 Miniature Schnauzers (including one known PMDS case) was genotyped for the AMHR2 mutation, and DNA from a single PMDS‐affected Belgian Malinois was sequenced for all coding exons of AMH and AMHR2. The Miniature Schnauzer cohort had an AMHR2 mutation allele frequency of 0.16 and a carrier genotypic frequency of 0.27. The genetic basis for PMDS in the Belgian Malinois was not determined, as no coding or splicing mutations were identified in either AMH or AMHR2. These findings support a benefit to AMHR2 mutation testing Miniature Schnauzers used for breeding or with cryptorchidism.     

Effect of broth from meat of linseed‐fed cattle on glucose‐stimulated insulin release in healthy male volunteers

Polyunsaturated fatty acid consumption has been shown to improve insulin sensitivity. We studied if administration of broth with beef meat enriched with polyunsaturated fatty acids influenced glucose‐stimulated insulin release in healthy male volunteers. Broth was made either from cattles undergone dietary supplementation with lightly bruised whole linseed in addition to feeding ad libitum on grass silage (test meal) or from those fed grass silage alone (control meal). Oral glucose tolerance tests (OGTT) were performed in patients after a 6‐day period of eating 300 ml broth containing 100 g meat once a day in addition to their otherwise normal mixed nourishment. During OGTT, blood samples were taken for blood glucose level and plasma insulin immunoreactivity before and 15, 30, 60, 90, 120, and 180 min after the glucose load. Glucose‐stimulated maximum increase in plasma insulin immunoreactivity was 42 ± 6.6 and 81 ± 7.4 mU/ml (p < 0.05) after the test and the control meals, respectively. However, both fasting and postload blood glucose levels were the same after either meal period. The results suggest an insulin‐sensitizing effect of food produced from beef cattle maintained on linseed diet in healthy human volunteers.     

Chlamydiaceae in riverine buffalo (Bubalus bubalis) and cows (Bos taurus) in Egypt with and without signs of reproductive disease

Abstract

AIMS: To obtain information and compare the prevalence of Chlamydiaceae in riverine buffalo (Bubalus bubalis) and cows (Bos taurus) in Egypt with and without clinical signs of reproductive disease.

METHODS: Vaginal swabs and blood samples were collected from animals attending Governmental Veterinary Clinics without (buffalo n=39, cows n=20) and with (buffalo n=63, cows n=53) signs of reproductive disease. Serum samples were tested for antibodies to Chlamydiaceae using complement fixation testing (CFT). Vaginal swabs were tested for Chlamydiaceae following inoculation into Vero cells and 6-day-old embryonated chicken eggs, using modified Giménez and immunoperoxidase staining, PCR analyses targeting the omp2 gene, and Restriction Fragment Length Polymorphism PCR (RFLP-PCR) for species identification.

RESULTS: Antibodies to Chlamydiaceae were detected in 30/39 (77%) and 50/63 (79%) buffalo without and with signs of reproductive disease, respectively, and 10/20 (50%) and 39/53 (74%) of cows with and without signs of reproductive disease, respectively. Positive samples from PCR analysis were identified in 31/39 (79%) and 37/63 (59%) buffalo without and with signs of reproductive disease, respectively, and 12/20 (60%) and 46/53 (89%) of cows without and with signs of reproductive disease, respectively. Using RFLP-PCR, 57/68 (84%) of samples from buffalo, and 47/58 (81%) from cows, were identified as Chlamydophila psittaci and the reminder as Cp. abortus. From the CFT and PCR results there was no significant difference in the prevalence of positive samples between species, or between animals without or with signs of reproductive disease.

CONCLUSION: The presence of anti-Chlamydiaceae antibodies in 77% of the animals with signs of reproductive disease and the detection of Chlamydiaceae in 72% of vaginal swabs of the animals suggest a pathogenic role by Chlamydiaceae in riverine buffalo and cows. The main Chlamydiaceae found in the genital tract of cattle in Egypt were Cp. psittaci and Cp. abortus.

CLINICAL RELEVANCE: Chlamydophila spp. should be included in diagnostic algorithms for reproductive disorders, in order to assess the real burden of Chlamydophila associated disease in buffalo and cattle and to evaluate the potential value of vaccines.     

Pregnancy status determination in mares using a rapid lateral flow test for measuring serum oestrone sulphate

AIMS: To develop a means of determining pregnancy status in horses based on measuring serum oestrone sulphate (OS) concentrations using a rapid lateral flow immunoassay, and to determine the assay's effectiveness using a visual end-point.

METHODS: Serum samples from mares >100 days post-mating (n=701) were assayed using a nitrocellulose membrane-based lateral flow immunoassay device. The device was developed using membrane-bound 1,3,5 (10)-estratrien-3-ol-17-one conjugated to bovine serum albumin as the capture antigen, and an OS-detection monoclonal antibody coupled to colloidal gold as the visible detection reagent. Concentrations of the coating antigen and OS monoclonal antibody were optimised so that the working range would allow pregnancy status to be determined from a visual end-point. The test was run by adding 0.1 ml serum to the sample well of a plastic cassette encasing the test membrane. As the serum migrated along the membrane, a test dot and control line were generated on it within 5–10 min. The intensity of the test dot was inversely proportional to the concentration of OS in the serum sample being tested. Results were compared with those from a validated OS enzyme immunoassay (EIA) and subsequent foaling or return to oestrus of the mares.

RESULTS: Serum samples with OS concentrations <10 ng/ml, indicative of non-pregnancy in mares >100 days post-mating, generated a test end-point consisting of a highly visible test dot and control line, whereas serum OS concentrations >50 ng/ml, indicative of pregnancy, generated a control line only. The test correctly identified 384/389 (98.7%) non-pregnant mares tested, and 303/312 (97.1%) pregnant mares tested that were >100 days post-mating. The lateral flow test devices were stable for at least 12 months when stored at 4°C, sealed in aluminium pouches with desiccant.

CONCLUSION: This novel, rapid, easy-to-use, lateral flow immunoassay offers a practical alternative to traditional laboratory-based immunoassays for measuring serum OS concentra- tions in mares for determining their pregnancy status.     

Effect of Feeding and Body Condition Score on Multiple Ovulation and Embryo Production in Zebu Cows

The aim of this study was to standardize the feeding regimen and the body condition score (BCS) for maximum superovulatory responses in indigenous zebu cows. Ten regularly cycling 5–8‐year‐old dry cows, weighing 176–260 kg with BCS 2.5–4.5 were divided into two equal groups at random. The groups were maintained on either a good‐nutrition or a high‐nutrition diet. The feedstuffs were analysed by proximate feed analysis and the metabolizable energy content was estimated. After 3 months feeding, individual cows were injected (i.m) with 1500 IU pregnant mare's serum gonadotrophin (PMSG) at day 10 or day 11 of the oestrous cycle (day of oestrus = day 0). Alfaprostol (6 mg) was injected (i.m) 48 h after the injection of PMSG to induce oestrus. At day 6 or day 7 (day of insemination = day 1), the contents of individual uterine horns were flushed with 150–200 ml of phosphate‐buffered saline + 0.2% bovine serum albumin using a two‐way Foley catheter. The embryos were identified, evaluated and graded as excellent, good, fair or poor under a stereomicroscope. For the good‐ and high‐nutrition diets, the daily intake of green grass, straw, concentrate, dry matter, crude protein and estimated metabolizable energy by individual cows were 5 and 6 kg, 3 and 3 kg, 1.5 and 3.5 kg, 4.87 and 6.82 kg, 0.39 and 0.74 kg, and 39.60 and 59.12 MJ, respectively. The protein content was 8 and 11% in the good‐ and high‐nutrition diets, respectively. The two groups of cows on different nutritional diets differed significantly with regard to body weight, body condition score and number of palpated corpora lutea (p < 0.01). For cows on the good‐nutrition diet, the median number of recovered embryos and transferable quality embryos were three and two, respectively. The recovery rate of embryos was 79.30% of palpated corpora lutea. Cows on the high‐nutrition diet did not yield any embryos. The indigenous zebu cows fed on the good‐nutrition diet with BCS 2.5–3 were considered suitable for the induction of superovulation, the cows on the high‐nutrition diet with BCS 4–4.5 were unsatisfactory and were more prone to cyst formation in the ovaries.     

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.     

Effects of Protein Content and Composition on White Noodle Making Quality: Color

Wheat cultivars, representing three winter and three spring wheats were grown in western Canada with six levels of nitrogen fertilizer and flours were prepared from them with an extraction rate of 65%. Using a chromameter, flour color and the color of uncooked white noodle sheets made from these flours with different resting times were assessed. The cooked noodle sheet color was also assessed. While protein content initially declined with added nitrogen and increased with further nitrogen addition, brightness (L*) of flour decreased and redness (a*) and yellowness (b*) increased. Positive correlation coefficients of flour brightness with particle size index (PSI) were also observed. Flour redness (a*) and yellowness (b*) were also affected by flour moisture content, whereas L* values were not significantly correlated with moisture contents. For the uncooked white noodle sheet, as protein content increased brightness decreased but there was an increase in a* and b* values. Thus, the L* value for noodle sheets was negatively correlated with the a* and b* values. The percentages of monomeric protein and soluble glutenin in flour were equal to or better than protein content in relation to most noodle sheet color characters. Uncooked noodle sheet brightness decreased, while redness and yellowness increased with rest time. In general, uncooked white noodle sheets prepared from different wheat flours can be ranked in terms of brightness and yellowness within each level of nitrogen fertilization.     

Breeding durum wheat for pasta quality in Canada

Canada was the largest durum wheat (Triticum turgidum var durum) producer in 1994, and in recent years supplied over 70% of world export trade in durum. Breeding for pasta quality is, therefore, a primary objective in Canadian durum breeding programs. Control of cultivar registration and stringent grading standards ensure that durum of consistent high quality is produced for domestic and export markets. The objectives of breeding programs include: improvement of traits related to production concerns, such as grain yield, disease resistance and sprouting resistance, and those related to end-use quality, such as protein concentration and quality; milling quality factors, such as semolina yield; colour of the wheat, semolina and pasta; and cooking quality. Selection and testing for quality begins at very early generations and becomes more stringent for advanced inbred lines. Selection is practised at the F₁ or F₂, where appropriate, using monoclonal antibodies to identify desirable gamma gliadins (γ-45 ) or low molecular weight glutenin subunits (LMW 2) that have been shown to be related to end-use quality. Grain from early generation yield trials, starting at F₄, is screened for protein concentration and pigment content by Near Infrared reflectance, and for gluten strength by sodium dodecyl sulphate (SDS) sedimentation and micro-mixograph. Promising lines entered into multi-location yield trials are screened with more time-consuming procedures to fully assess suitability for pasta processing. These tests include semolina yield, ash and colour, and predictions of gluten strength such as mixograph and alveograph, and cooking quality. Candidate cultivars with quality equal to or better than the mean of the check cultivars can be proposed for registration after three years in the Durum Cooperative Test. It takes approximately 10 years from performing a cross to registrating a new cultivar. This revised version was published online in August 2006 with corrections to the Cover Date.     

The percolation phase transition in sea Ice

Sea ice exhibits a marked transition in its fluid transport properties at a critical brine volume fraction pc of about 5 percent, or temperature Tc of about -5 degreesC for salinity of 5 parts per thousand. For temperatures warmer than Tc, brine carrying heat and nutrients can move through the ice, whereas for colder temperatures the ice is impermeable. This transition plays a key role in the geophysics, biology, and remote sensing of sea ice. Percolation theory can be used to understand this critical behavior of transport in sea ice. The similarity of sea ice microstructure to compressed powders is used to theoretically predict pc of about 5 percent.     

Using RapidEye imagery to identify within-field variability of crop growth and yield in Ontario,Canada

Precision Agriculture - Remote sensing has been recognized as a cost-effective way to detect the spatial and temporal variability of crop growth and productivity. In this study, multispectral...