Retinol, β-carotene and β-tocopherol concentrations in mare and foal plasma and in colostrum

Fat-soluble vitamins transfer poorly across the diffuse epitheliochorial placenta of the mare, so the neonatal foal is dependent on its supply via colostrum. Concentrations of retinol (RT), β-carotene (BC), and β-tocopherol (AT) were assayed in samples of plasma, colostrum and milk from nine mares of mixed light breeding and their foals at parturition and days 1, 2, and 4 postpartum. Samples were analyzed simultaneously for RT, BC, and AT using a new, less time-consuming reverse phase high-pressure liquid chromatography (HPLC) method. Feeds were analyzed, and mean daily intakes calculated. Mare plasma RT increased from day 1 to day 4 (P = .033), and plasma AT declined linearly from day 0 to day 4 (P = .006). Colostrum concentrations of all vitamins increased from parturition to day 1, and then declined rapidly over the 4 days. Foal plasma BC increased from parturition to day 4 (P = .080), and plasma AT increased from parturition to day 2 (P < .001), and 4 (P = .060). These observations suggest that three times the current recommendation of vitamin A is sufficient for pregnant mares, in keeping with previous studies of growth, pregnancy, and lactation. In contrast, the linear decrease in mare plasma AT suggests that 1.2 times the current recommendation of vitamin E may be less than optimal for pregnant mares.

Introduction

The diffuse epitheliochorial placenta of the mare does not allow fat-soluble vitamins to cross with ease to the fetus,[1] so their status in the neonatal foal is dependent on colostrum. This first milk contains these vitamins, which are necessary for many functions.Vitamin A is required for vision, reproduction, growth, development, and maintenance of epithelial cells, and osteoclast activity in bone formation. In addition, vitamin A deficiencies reduce resistance to disease, can cause neurological degeneration, and congenital defects.[2] Vitamin E, an antioxidant, prevents lipid peroxidation and cell membrane damage. Its deficiency in horses has been associated with a form of muscular dystrophy and impaired immune function. [3]Studies on the mare and foal have described blood concentrations of one or two of these vitamins,[4, 5 and 6] but only one involved all three. [7] In that study, vitamin concentrations were assayed in mare blood and milk, but only once in foal blood at parturition, so data during the periparturant period is incomplete.The objectives of this enumerative study were to determine concentrations of RT, BC, AT in plasma and colostrum, to evaluate colostral transfer of these vitamins, to assess the adequacy of vitamin intakes of mares, and to develop a time-saving single-run HPLC method.

Materials and methods

Animals

Nine healthy aged multiparous mares of mixed light breeding (13.6 ± 1.5 years, 592 ± 20 kg body mass) and their foals were sampled. The institutional animal care and use committee approved the protocol. Mares were kept on meager late-winter mixed grass pasture, with free access to round bale orchard and fescue grass hay, water, and trace mineral/vitamin salt. They were fed 2 kg of concentrate twice daily at 7:00 and 3:00 . The diet met or exceeded current recommendations for pregnant mares at 11 months gestation for energy.[8] Samples of hay and concentrate were submitted for partial proximate analysis to the Virginia Tech Forage Testing Laboratory where standard AOAC [9] methods are used ( Table 1). Samples of feeds were also taken for analysis of RT, BC, and AT ( Table 2).     

Fat digestibility in Equus caballus follows increasing first-order kinetics

The digestibility of ether extract varies greatly from forages to grains and further to added fats consisting mainly of triglycerides. This variation has been attributed to two main factors, the presence of nonhydrolyzable substances in the ether extract, especially in leafy foods, and the dilution of endogenous fecal fat. A compilation of results from 188 equine digestion balance observations on five basal feeds and 18 test feeds with added fats demonstrated a true digestibility of fat approaching 100% and an endogenous fecal fat of 0.22 g x d(-1) x kg BW(-1). The results revealed that nonhydrolyzable ether extract and endogenous fecal fat were insufficient to account for the difference between true digestibility and apparent digestibilities of ether extract in basal feeds and partial digestibilities of added fats in test feeds. A third possible contributing factor was demonstrated: an increasing first-order relationship between observed digestibilities (D, %) and the fat content of the feed (F, g/kg): D = 92.0 - 92.0e(-F/342). r2 = 0.81, P < 0.001. This equation indicates that 46% digestibility (half maximum) occurs at an ether extract or fat content of 24 g/kg, which is common in forages. It is consistent with fat digestibility or efficiency of absorption being a function of the rate of lipolysis, especially when residence time in the small intestine is limited. Consequently, we suggest that the kinetics of lipases, which are difficult to measure, may contribute to low digestibility when substrate concentration in the small intestine is low due to a low fat content in food. The status of vitamins A and E might be affected by low dietary fat contents and might be improved by fat supplementation.     

Insulin sensitivity decreases with obesity, and lean cats with low insulin sensitivity are at greatest risk of glucose intolerance with weight gain

This study quantifies the effects of marked weight gain on glucose and insulin metabolism in 16 cats which increased their weight by an average of 44.2% over 10 months. Significantly, the development of feline obesity was accompanied by a 52% decrease in tissue sensitivity to insulin and diminished glucose effectiveness. In addition, glucose intolerance and abnormal insulin response occurred in some cats. An important finding was that normal weight cats with low insulin sensitivity and glucose effectiveness were at increased risk of developing impaired glucose tolerance with obesity. High basal insulin concentrations or low acute insulin response to glucose also independently increased the risk for developing impaired glucose tolerance. Male cats gained more weight relative to females and this, combined with their tendency to lower insulin sensitivity and higher insulin concentrations, may explain why male cats are at greater risk for diabetes. Results suggest an underlying predisposition for glucose intolerance in some cats, which is exacerbated by obesity. These cats may be more at risk of progressing to overt type 2 diabetes mellitus.     

Mucopolysaccharidosis type VI in a Miniature Poodle-type dog caused by a deletion in the arylsulphatase B gene

Abstract

AIM: To investigate and characterise an inborn error of metabolism in a dog with skeletal and ocular abnormalities.

METHODS: A 2.5-year-old small male Miniature Poodle-like dog was presented with gross joint laxity and bilateral corneal opacities. Clinical examination was augmented by routine haematology, serum chemistry, radiographs, pathology, enzymology and molecular genetic studies. Euthanasia was requested when the dog was 3 years of age because of progressively decreasing quality of life.

RESULTS: Radiology revealed generalised epiphyseal dysplasia, malformed vertebral bodies, luxation/subluxation of appendicular and lumbosacral joints with hypoplasia of the odontoid process and hyoid apparatus. These clinical and radiographic findings, together with a positive urinary Berry spot test for mucopolysaccharides, and metachromatic granules in leucocytes, were indicative of a mucopolysaccharidosis (MPS), a lysosomal storage disease. Histological lesions included vacuolation of stromal cells of the cornea, fibroblasts, chondrocytes, macrophages and renal cells. The brain was essentially normal except for moderate secondary Wallerian-type degeneration in motor and sensory tracts of the hind brain. Dermatan sulphate-uria was present and enzymology revealed negligible activity of N-acetylgalactosamine-4-sulphatase, also known as arylsulphatase B, in cultured fibroblasts and liver tissue. A novel homozygous 22 base pair (bp) deletion in exon 1 of this enzyme's gene was identified (c.103_124del), which caused aframe-shift and subsequent premature stop codon. The “Wisdom pure breed-mixed breed” test reported the dog as a cross between a Miniature and Toy Poodle.

CONCLUSIONS: The clinicopathological features are similar to those of MPS type VI as previously described in dogs, cats and other species, and this clinical diagnosis was confirmed by enzymology and molecular genetic studies. This is an autosomal recessively inherited lysosomal storage disease.

CLINICAL RELEVANCE: The prevalence of MPS VI in Miniature or Toy Poodles in New Zealand and elsewhere is currently unknown. Due to the congenital nature of the disorder, malformed pups may be subject to euthanasia without investigation and the potential genetic problem in the breed may not be fully recognised. The establishment of a molecular genetic test now permits screening for this mutation as a basis to an informed breeding policy.     

Descriptive epidemiological study of the use of antimicrobial drugs by companion animal veterinarians in New Zealand

Abstract

AIM: To describe the patterns of use of antimicrobial drugs by veterinary surgeons treating commonly presented bacterial infections in companion animals in New Zealand.

METHODS: A postal survey of 800 randomly selected companion animal veterinarians practicing in New Zealand was conducted between August and December 2008. Data were collected regarding the antimicrobials prescribed for recent cases of skin, ear and urinary tract infections; the use of culture and susceptibility testing; and veterinarian characteristics such as proportion of time spent treating companion animals and recent attendance at continuing professional development (CPD) events. Potential associations within the data were explored using extended mosaic plots and multivariable regression models.

RESULTS:Completed surveys from 393 respondents were available for analysis, providing data on systemic antimicrobial drug use for 1,799 cases of presumptive bacterial infections. The most frequently prescribed drugs were amoxicillin-clavulanic acid (864 cases, 48%), cephalexin (558, 31%), and fluoroquinolones (198, 11%). Of 359 cases of canine superficial pyoderma, 157 (44%) were treated with amoxicillin-clavulanic acid and 155 (43%) were treated with cephalexin with median reported treatment durations of 7 and 10 days, for these two drugs respectively. Culture and susceptibility tests had been used in 376 of 1,984 (19%) of all reported cases and 160 (43%) of these were suspected urinary tract infections. Practitioners that spent 100% of their time treating companion animals and who had attended a CPD course related to companion animals within the 12 months prior to completing the survey were more likely to submit a sample for culture and susceptibility testing and to prescribe longer courses of antimicrobials for the treatment of canine pyoderma than practitioners who spent less than 100% of their time treating companion animals and had not attended a CPD course in the last 12 months.

CONCLUSIONS: Broad-spectrum drugs considered by the World Health Organisation to be critically important for human health, such as fluoroquinolones and amoxicillin-clavulanic acid, are amongst the most frequently prescribed antimicrobials in companion animal medicine, and these drugs are often prescribed without submitting a sample for culture and susceptibility testing.

CLINICAL RELEVANCE: Many cases of superficial pyoderma were treated for less than the recommended duration of 21 days, which may contribute to a higher rate of recurrent pyoderma and the development of drug resistance within the causal bacteria. Veterinarians should be aware that the use of fluoroquinolones, in particular, should be based upon the results of a culture and susceptibility test.     

Prostaglandin Endoperoxide Synthase 2 (PTGS2) and Prostaglandins F2α and E2 Synthases (PGFS and PGES) Expression and Prostaglandin F2α and E2 Secretion Following Oestrogen and/or Progesterone Stimulation of the Feline Endometrium

Sex steroids in synergy with prostaglandins (PG) are involved in the regulation of cyclic ovarian function. In this study, we investigated the mRNA expression of three genes involved in arachidonic acid (AA) metabolism and hence PG production in domestic cats: PG‐endoperoxide synthase (PTGS2), PGF_2α synthase (PGFS) and PGE₂ synthase (PGES). Feline endometria (n = 16) were collected at oestrus and mid and late phases of pseudopregnancy. In addition, the effects of E₂ and/or P₄ on PG secretion and gene expression on endometrial explants were studied in an in vitro culture system. Expression levels of all examined genes were up‐regulated at the mid phase of pseudopregnancy. The effects of E₂ and/or P₄ treatment on both PG secretion and expression of the genes were observed after 12 h of culture. Expression of PGES was significantly up‐regulated by E₂ plus P₄ at oestrus and the mid phase of pseudopregnancy and was also up‐regulated by a single treatment with P₄ at late pseudopregnancy (p < 0.05). Simultaneous incubation with E₂ and P₄ up‐regulated PTGS2 gene expression at oestrus and mid‐luteal phase (p < 0.05). Progesterone plus E₂ significantly increased PGE₂ secretion at oestrus and the mid phase of pseudopregnancy. However, treatment with E₂ and/or P₄ affected neither PGF_2α secretion nor PGFS expression at any phase after 12 h of culture. The overall findings indicate that genes involved in PG synthesis are up‐regulated at the mid phase of pseudopregnancy. An increase in PGE₂ secretion and up‐regulation of PGES and PTGS2 are the main responses of the endometrium to treatment with E₂ and P₄ at oestrus and the mid phase of pseudopregnancy in the cat. These data support the hypothesis that ovarian sex steroids via endometrial PGE₂ are involved in endocrine homoeostasis, especially at oestrus and the mid, but not the late, phase of pseudopregnancy in cats.     

A Case of Successful Pregnancy in a Ewe with Uterus Didelphys

Uterus didelphys is a rare congenital abnormality of the reproductive tract. Although it occurs in various species, there are no published reports describing pregnancy outcome in association with this abnormality. Herein we describe a case of successful unilateral singleton pregnancy in a ewe incidentally found to have uterus didelphys during the course of a biomedical research study. The pregnancy was established using assisted reproductive techniques and interrupted in late gestation, at which point the abnormality was identified. Serial ultrasound assessment of foetal biometry revealed a normal foetal growth trajectory. Despite a 45% reduction in placentome number, total placentome weight was near normal secondary to compensatory placentome growth and development. To our knowledge, this is the first detailed report of normal foetal growth in an animal with uterus didelphys and illustrates the ability of the ovine placenta to adapt to a reduced number of placentomes and maintain foetal nutrient supply.