Random-effects linear model application to herd-level assessment of bovine hepatic trace mineral concentrations.

To evaluate the utility of random-effects linear modeling for herd-level evaluation of trace mineral status, we performed a retrospective analysis of the results for trace mineral testing of bovine liver samples submitted to the Michigan State University Veterinary Diagnostic Laboratory between 2011 and 2017. Our aim was to examine random-effects models for their potential utility in improving interpretation with minimal sample numbers. The database consisted of 1,658 animals distributed among 121 herds. Minerals were assayed by inductively coupled plasma-mass spectroscopy, and included cobalt, copper, iron, molybdenum, manganese, selenium, and zinc. Intraclass correlation coefficients for each mineral were significantly different (p < 0.001) from zero and ranged from 0.38 for manganese to 0.82 for selenium, indicating that the strength of herd effects, which are presumably related to diet, vary greatly by mineral. Analysis of the distribution and standard errors of best linear unbiased predictor (BLUP) values suggested that testing 5-10 animals per herd could place herds within 10 percentile units across the population of herds with 70-95% confidence, the confidence level varying among minerals. Herd means were generally similar to BLUPs, suggesting that means could be reasonably compared to BLUPs with respect to the distributions reported here. However, caution in interpreting means relative to BLUPs should be exercised when animal numbers are small, the standard errors of the means are large, and/or the values are near the extremes of the distribution.

Metformin effectively restores the HPA axis function in diet-induced obese rats.

INTRODUCTION: The hypothalamo-pituitary-adrenal (HPA) axis is perturbed in obesity. We previously reported presence of leptin resistance in the brainstem and uncoupling between central noradrenergic tone and the HPA axis in obesity-prone (DIO) rats. Metformin is shown to lower body weight and adiposity, but the underlying mechanism is unclear. We hypothesized that this is associated with restored HPA axis function. METHODS: Adult male DIO rats were placed on either a regular chow or HF diet for 7 weeks. Starting week 4, the animals were given either a low dose (60 mg/kg) or high dose (300 mg/kg) of metformin in drinking water. In addition to body weight and feeding, we examined different arms of the HPA axis to test if metformin can reinstate its function and coupling. To understand potential mechanisms, leptin signaling in the brainstem and circulating free fatty acid levels were also assessed. RESULTS: Metformin treatment lowered weight gain, fat mass, caloric intake, and serum leptin levels. HPA axis activity as determined by corticotropin-releasing hormone in the median eminence and serum corticosterone was decreased by metformin in a dose-dependent manner, and so was norepinephrine (NE) in the paraventricular nucleus. Importantly, metformin completely normalized the NE-HPA axis uncoupling. While brainstem pSTAT-3 and SOCS-3, key markers of leptin signaling, were not different between groups, circulating saturated and unsaturated free fatty acids were reduced in HF-fed, metformin-treated animals. CONCLUSIONS: These findings suggest that oral metformin can successfully correct HPA axis dysfunction that is associated with lowered circulating free fatty acids in DIO rats, thereby uncovering a novel effect of metformin in the treatment of obesity.

Serum retinol, ß-carotene, and α-tocopherol as biomarkers for disease risk and milk production in periparturient dairy cows.

The effectiveness of using serum vitamin concentrations as biomarkers to predict diseases in dairy cows during the periparturient period is not well known. The objective of this study was to evaluate the association between serum ß-carotene, retinol, and α-tocopherol concentrations and periparturient cow diseases in commercial dairies. We measured serum concentrations of these vitamin-active compounds at dry-off and during close-up (approximately 3 wk before calving) and early lactation (approximately 7 d post-calving), and we examined their association with clinical diseases in the first 30 d in milk. Diseases were diagnosed by trained personnel and recorded using database software. Blood samples were taken from 353 cows from 5 different farms over a 3-yr period. Blood samples were analyzed for ß-carotene, retinol, α-tocopherol, and cholesterol. We built separate mixed logistic regression models for each disease outcome: hyperketonuria, lameness, mastitis, uterine diseases (retained placenta or metritis), and an aggregate outcome. For the aggregate outcome, a cow was considered positive if she had one or more of the following: hyperketonuria, lameness, mastitis, uterine disease, pneumonia, milk fever, or displaced abomasum. Concentrations of all 3 fat-soluble vitamins decreased significantly in early lactation relative to the 2 prepartum sampling times. Serum retinol concentrations at close-up and early lactation were negatively associated with odds of developing postpartum hyperketonuria. At early lactation, cows with uterine disease had lower serum retinol concentrations than cows without uterine disease. Similarly, lower serum retinol concentrations were associated with greater odds of having any one disease in the aggregate outcome. First-test 305-d mature-equivalent milk yield was positively correlated with increased serum α-tocopherol and negatively correlated with ß-carotene concentrations. This study demonstrates the potential for serum ß-carotene, retinol, and α-tocopherol to serve as biomarkers for disease risk.

Oleic acid stimulates monoamine efflux through PPAR-α: Differential effects in diet-induced obesity.

Obesity continues to be a growing health concern around the world, and elevated levels of free fatty acids as a result of high-fat intake might play a role in neuroendocrine alterations leading to obesity. However, it is unclear how fatty acids affect neuroendocrine functions and energy metabolism. Since hypothalamic monoamines play a crucial role in regulating neuroendocrine functions relating to energy balance, we investigated the direct effects of oleic acid on hypothalamic monoamines and hypothesized that oleic acid would activate peroxisome proliferator-activated receptor alpha (PPAR-α), a nuclear transcription factor involved with fatty acid metabolism, to affect monoamines. We also hypothesized that this response would be subdued in diet-induced obesity (DIO). To test these hypotheses, hypothalami from Sprague Dawley and DIO rats were incubated with 0 (Control), 0.00132 mM, 0.132 mM, 1.32 mM oleic acid, 50 µM MK 886 (a selective PPAR- α antagonist), or oleic acid + MK 886 in Krebs Ringers Henseleit (KRH) solution. HPLC-EC was used to measure monoamine levels in perfusates. Oleic acid produced a significant increase in norepinephrine, dopamine, and serotonin levels in a dose-dependent manner, and incubation with MK886 blocked these effects. The effect of oleic acid on hypothalamic monoamines was attenuated in DIO rats. These findings suggest that PPARα probably plays an essential role in fatty acid sensing in the hypothalamus, by affecting monoamine efflux and DIO rats are resistant to the effects of oleic acid.

Effects of Super Nutritional Hepatic Copper Accumulation on Hepatocyte Health and Oxidative Stress in Dairy Cows.

Concerns regarding excessive hepatic copper concentrations in dairy cows have increased. The objective of this study was to determine the association of hepatic copper concentrations with evidence of liver disease. Blood and liver samples were collected at the time of slaughter in cull dairy cows (n=100). Liver samples were analyzed for copper using inductively coupled plasma mass spectrometry and crude fat using liquid-liquid extraction and gravimetry. Serum samples were analyzed for glutamate dehydrogenase, γ-glutamyltransferase, sorbitol dehydrogenase, aspartate aminotransferase activities, and bile acid concentrations. Liver samples were examined histologically for inflammation, fibrosis, and rhodanine staining. Animals were stratified by hepatic copper concentration and samples in the highest and lowest quintiles (Q5 and Q1) were evaluated for oxidative stress. Systemic indices of oxidative stress included serum reactive oxygen and nitrogen species (RONS) and total antioxidant potential (AOP). Tissue-level oxidative stress was assessed by immunohistochemistry using 4-hydroxynonenal (4HNE) and 3-nitrotyrosine (3NIT) stains to score the relative abundance and distribution of oxidized lipid and protein products, respectively. Mean hepatic copper concentration was 496.83 µg/g and median 469.72 µg/g and ranged from 70.56 to 1264.27 µg/g dry tissue. No association was found between hepatic copper concentrations and clinicopathological or histological evidence of hepatic damage or dysfunction. There was a significant increase in the amount of IHC staining of 4HNE and 3NIT in Q5 compared with Q1. Moreover, the IHC staining mirrored the distribution of the copper-specific stain rhodanine. These results demonstrate that cows with elevated hepatic copper concentrations had no evidence of active liver disease but had increased hepatic oxidative stress.

Short communication: Survey of hepatic copper concentrations in Midwest dairy cows.

Previous research from our laboratory and others indicates that liver copper concentrations in dairy cattle are commonly well above those recognized as adequate for the nutritional needs of the animal. It has also been speculated that hepatic copper concentrations have been increasing in recent years. Unlike other species, the threshold at which elevated liver copper concentrations becomes deleterious to hepatocytes is not known for cattle. Therefore, the objectives of this study were 3-fold: (1) to delineate differences in the range and mean dry matter hepatic copper concentration for dairy cattle in a retrospective analysis (January 1, 2007, to December 31, 2015), (2) to investigate hepatic copper concentrations in Midwest cull dairy cattle, and (3) to evaluate histologic changes in hepatocellular morphology in the context of copper concentration in cull cows. Furthermore, microscopic changes in hepatocellular morphology or architecture were examined and scored for evidence of inflammation, fibrosis, necrosis, and abundance of rhodanine-stained granules using hematoxylin and eosin and rhodanine staining. The retrospective analysis found copper concentrations within a range of 3 to 1,963 µg/g, with a mean of 473 µg/g. Hepatic copper concentrations in our retrospective study did not increase with time. In our abattoir analysis, copper concentrations ranged from 15 to 978 µg/g, with a mean of 390 µg/g. This study found that the range and mean hepatic copper concentrations were comparatively less in the current abattoir study than copper concentrations in our retrospective analysis. There was no evidence for hepatocellular changes associated with increased copper burdens in this study population.

Influence of various concentrate-to-roughage ratios on dietary intake and nutrient digestibilities of weanling horses.

The objective of this study was to compare the feed intake and the apparent digestibilities of three different diets varying in concentrate-to-roughage ratios in weanling horses (n = 24) at 5 and 8 months of age. Horses were stratified by breed, gender, birth date and body weight and assigned to one of three dietary treatments containing the following concentrate-to-roughage ratios on an as-fed basis: 70:30 (High Con), 50:50 (Equal) and 30:70 (Low Con). All horses were fed their respective diets for a 10-day adaptation period and a 4-day collection period at 5 and 8 months. There were no differences in BW or daily feed intake among treatments during both trials. The horses consuming Low Con had a greater amount of faecal output than High Con at both 5 and 8 months (p < 0.01). At 5 months, High Con had the highest crude protein (CP) digestibility (p < 0.05). At 8 months, High Con had a higher CP digestibility than Low Con (p < 0.01) and tended to be higher than Equal (p = 0.07). Acid detergent fibre (ADF) digestibility did not differ among treatments; however, horses fed the Low Con tended to digest a higher percentage of neutral detergent fibre (NDF) than both the Equal and High Con treatments (p = 0.09). Horses in the High Con treatment tended to digest a higher percentage of energy than those in the Low Con treatment (p = 0.06). Weanlings seem to digest protein more thoroughly when fed high-concentrate diets and may digest fibre more efficiently when fed diets higher in fibre.

Pharmacologic evaluation of ammonium tetrathiomolybdate after intravenous and oral administration to healthy dogs.

OBJECTIVE: To evaluate pharmacokinetics of ammonium tetrathiomolybdate (TTM) after IV and oral administration to dogs and effects of TTM administration on trace mineral concentrations. ANIMALS: 8 adult Beagles and Beagle crossbreds (4 sexually intact males and 4 sexually intact females). PROCEDURES: Dogs received TTM (1 mg/kg) IV and orally in a randomized crossover study. Serum molybdenum and copper concentrations were measured via inductively coupled plasma mass spectrometry in samples obtained 0 to 72 hours after administration. Pharmacokinetics was determined via noncompartmental analysis. RESULTS: For IV administration, mean ± SD terminal elimination rate constant, maximum concentration, area under the curve, and half-life were 0.03 ± 0.01 hours(-1), 4.9 ± 0.6 µg/mL, 30.7 ± 5.4 µg/mL•h, and 27.7 ± 6.8 hours, respectively. For oral administration, mean ± SD terminal elimination rate constant, time to maximum concentration, maximum concentration, area under the curve, and half-life were 0.03 ± 0.01 hours(-1), 3.0 ± 3.5 hours, 0.2 ± 0.4 µg/mL, 6.5 ± 8.0 µg/mL•h, and 26.8 ± 8.0 hours, respectively. Oral bioavailability was 21 ± 22%. Serum copper concentrations increased significantly after IV and oral administration. Emesis occurred after IV (2 dogs) and oral administration (3 dogs). CONCLUSIONS AND CLINICAL RELEVANCE: Pharmacokinetics for TTM after a single IV and oral administration was determined for clinically normal dogs. Absorption of TTM after oral administration was variable. Increased serum copper concentrations suggested that TTM mobilized tissue copper. Further studies will be needed to evaluate the potential therapeutic use of TTM in copper-associated chronic hepatitis of dogs.

Hyperplastic goiter in two adult dairy cows.

Iodine excess and resultant hyperplastic goiter are well documented in neonatal ruminants, but little is reported on iodine excess in adult ruminants and associated histological changes of the thyroid gland. Two adult Holstein cows from a Michigan dairy herd that had lost several other animals had nonspecific clinical signs of illness and were submitted for necropsy. Thyroid glands of one of these 2 animals were grossly and markedly enlarged, and histologically, thyroid glands from both animals had regions of cystic nodular hyperplasia and follicular atrophy. Thyroid glands from both animals had markedly elevated iodine concentrations. Investigation into the potential source of excessive iodine on the farm revealed multiple sources of supplemental dietary iodine and probable uneven feed and mineral mixing. Based on the findings of this investigation, adult cattle could be susceptible to excessive doses of iodine. Possibility of previous iodine deficiency before supplementation period, with subsequent development and persistence of thyroid hyperplasia and cystic change, cannot be completely excluded. Current findings suggested that iodine excess in adult cattle can result in nodular hyperplastic goiter. Use of iodized salt in mineral supplements in adult dairy herds is common practice, and accidental excessive iodine supplement may be more common than reported. Recognizing gross and histological thyroid gland changes, consisting of concurrent cystic follicular hyperplasia, atrophy, and fibrosis should raise suspicion of iodine excess and/or prior deficiency in a cattle herd, and ancillary tests such as serum iodine measurements should be part of the diagnostic workup in suspected cases.

The use of blood analysis to evaluate trace mineral status in ruminant livestock.

This article summarizes effects and evaluation of 8 trace minerals considered significant in ruminant nutrition, both for nutritional deficiencies as well as production-related toxicosis: cobalt, copper, iron, iodine, manganese, molybdenum, selenium, and zinc. Changes in availability, metabolism, and amounts needed for optimum health and productivity in animals are their major effect; frank clinical toxicosis or severe nutritional deficiency are of limited concern in modern production agriculture. The information provided in this article can help to manage the risk of subtle effects that may alter performance and lifetime productivity.

Cooperating with industry to teach food-animal medicine.

Cooperative programs with agribusiness offer opportunities for colleges of veterinary medicine to expand their instructional programs in agricultural practice. Agribusinesses associated with livestock agriculture recognize the importance of veterinary medicine in maintaining a vibrant and successful industry. Stewardship of corporate support involves close communication with advocates within the companies, providing them with documentation of the potential effects of corporate investments. This article describes the creation of the Michigan State University (MSU) Training Center for Dairy Professionals, a key aspect of which was the identification of innovative and productive areas of mutual interest and benefit. In addition to supporting the dairy industry by training veterinary students, the program offers specific benefits to investors, including the use of MSU facilities and direct participation in veterinary instruction.

Pharmacokinetics of subcutaneous selenium in adult llamas.

Selenium (Se) deficiency disease has been described in camelids and only clinical data is available for administration of parenteral Se supplements. This study investigated the pharmacokinetic effects of subcutaneous Se injection (0.1 mg/kg) in llamas fed a diet adequate in Se. Absorption of Se was rapid with peak whole blood Se concentration at the first sampling time. Significant differences in whole blood Se concentration from before injection of Se were not found past 2 days after Se injection. Parenteral Se is unlikely to have a long-term effect on whole blood Se concentration in llamas fed adequate dietary Se.