The oral microbiome in alcohol use disorder: a longitudinal analysis during inpatient treatment.

BACKGROUND: Alcohol use disorder (AUD)-induced disruption of oral microbiota can lead to poor oral health; there have been no studies published examining the longitudinal effects of alcohol use cessation on the oral microbiome. AIM: To investigate the oral microbiome during alcohol cessation during inpatient treatment for AUD. METHODS: Up to 10 oral tongue brushings were collected from 22 AUD patients during inpatient treatment at the National Institutes of Health. Alcohol use history, smoking, and periodontal disease status were measured. Oral microbiome samples were sequenced using 16S rRNA gene sequencing. RESULTS: Alpha diversity decreased linearly during treatment across the entire cohort (P = 0.002). Alcohol preference was associated with changes in both alpha and beta diversity measures. Characteristic tongue dorsum genera from the Human Microbiome Project such as Streptococcus, Prevotella, Veillonella and Haemophilus were highly correlated in AUD. Oral health-associated genera that changed longitudinally during abstinence included Actinomyces, Capnocytophaga, Fusobacterium, Neisseria and Prevotella. CONCLUSION: The oral microbiome in AUD is affected by alcohol preference. Patients with AUD often have poor oral health but abstinence and attention to oral care improve dysbiosis, decreasing microbiome diversity and periodontal disease-associated genera while improving acute oral health.

The oral microbiome of patients undergoing treatment for severe aplastic anemia: a pilot study.

The microbiome, an intriguing component of the human body, composed of trillions of microorganisms, has prompted scientific exploration to identify and understand its function and role in health and disease. As associations between microbiome composition, disease, and symptoms accumulate, the future of medicine hinges upon a comprehensive knowledge of these microorganisms for patient care. The oral microbiome may provide valuable and efficient insight for predicting future changes in disease status, infection, or treatment course. The main aim of this pilot study was to characterize the oral microbiome in patients with severe aplastic anemia (SAA) during their therapeutic course. SAA is a hematologic disease characterized by bone marrow failure which if untreated is fatal. Treatment includes either hematopoietic stem cell transplantation (HSCT) or immunosuppressive therapy (IST). In this study, we examined the oral microbiome composition of 24 patients admitted to the National Institutes of Health (NIH) Clinical Center for experimental SAA treatment. Tongue brushings were collected to assess the effects of treatment on the oral microbiome. Twenty patients received standard IST (equine antithymocyte globulin and cyclosporine) plus eltrombopag. Four patients underwent HSCT. Oral specimens were obtained at three time points during treatment and clinical follow-up. Using a novel approach to 16S rRNA gene sequence analysis encompassing seven hypervariable regions, results demonstrated a predictable decrease in microbial diversity over time among the transplant patients. Linear discriminant analysis or LefSe reported a total of 14 statistically significant taxa (p < 0.05) across time points in the HSCT patients. One-way plots of relative abundance for two bacterial species (Haemophilus parainfluenzae and Rothia mucilaginosa) in the HSCT group, show the differences in abundance between time points. Only one bacterial species (Prevotella histicola) was noted in the IST group with a p value of 0.065. The patients receiving immunosuppressive therapy did not exhibit a clear change in diversity over time; however, patient-specific changes were noted. In addition, we compared our findings to tongue dorsum samples from healthy participants in the Human Microbiome Project (HMP) database and found among HSCT patients, approximately 35% of bacterial identifiers (N = 229) were unique to this study population and were not present in tongue dorsum specimens obtained from the HMP. Among IST-treated patients, 45% (N = 351) were unique to these patients and not identified by the HMP. Although antibiotic use may have likely influenced bacterial composition and diversity, some literature suggests a decreased impact of antimicrobials on the oral microbiome as compared to their effect on the gut microbiome. Future studies with larger sample sizes that focus on the oral microbiome and the effects of antibiotics in an immunosuppressed patient population may help establish these potential associations.

Flaxseed meal and oat hulls supplementation modulates growth performance, blood lipids, intestinal fermentation, bile acids, and neutral sterols in growing pigs fed corn-soybean meal-based diets.

The present study was conducted to determine the effect of flaxseed meal and oat hulls supplementation on growth performance, apparent total tract digestibility (ATTD) of fat, serum lipids, and concentrations of VFA, bile acids (BA), and neutral sterols (NS) in digesta and feces in growing pigs. Forty-eight Genesus [(Duroc boar × Yorkshire-Landrace sows] barrows (25.0 ± 0.32 kg initial BW) were housed in pairs. Pigs were assigned to 1 of the 3 corn-soybean meal-based diets-a basal corn-soybean meal-containing diet (control), a flaxseed meal-containing diet (FM), or an oat hulls-containing diet (OH)-in a completely randomized design. All diets were formulated to be isoenergetic and to contain similar standardized ileal digestible AA contents and meet other nutrient requirements for growing pigs. The experiment lasted for 28 d. Average daily feed intake; ADG; G:F; ATTD of fat, serum lipids, and digesta; and fecal VFA, BA, and NS concentrations were determined. Pigs fed the control or OH had greater final BW ( < 0.001), ADFI ( = 0.005), and ADG ( < 0.001) than FM-fed pigs. The ATTD of fat in the FM was lowest at 70.1% followed by 79.2% in OH and was greatest at 92.4% in the control ( = 0.020). Total serum cholesterol content was 2.25 and 1.99 mmol/L and lower ( < 0.001) in pigs fed FM and OH, respectively, than the 2.36 mmol/L in pigs fed the control. Pigs fed the FM and OH had greater ileal and cecal total VFA ( < 0.001), ileal deoxycholic acid ( < 0.01), and cecal ( < 0.001) and fecal cholesterol ( = 0.002) concentrations than those fed the control. Pigs fed the FM excreted more fecal lithocholic acid ( = 0.002) and ursodeoxycholic acid ( = 0.001) compared with those that consumed the control and OH. The concentrations of coprostanol in cecal digesta ( < 0.001) and feces ( = 0.011) were higher in pigs fed the FM and OH than in pigs fed the control. In conclusion, feeding flaxseed meal and oat hulls induced intestinal fermentation; however, the former depressed growth performance whereas the latter did not have any effect. Addition of flaxseed meal and oat hulls in growing pig diets reduced fat digestibility and serum cholesterol and stimulated malabsorption of primary BA and excretion of secondary BA and NS.

Surgical amputation of a digit and vacuum-assisted-closure (V.A.C.) management in a case of osteomyelitis and wound care in an eastern black rhinoceros (Diceros bicornis michaeli).

A 14-yr-old female eastern black rhinoceros (Diceros bicornis michaeli) presented with progressive suppurative osteomyelitis in her left hind lateral toe. beta-Hemolytic Streptococcus sp. was isolated. The animal was treated with multiple systemic antibiotics, and topical wound cleansing. Repeated debridements and nail trimmings were performed for 5 mo prior to electing amputation. The toe was surgically amputated under general anesthesia between the first and second phalanges. Analgesia was diffused into the wound topically via a catheter and elastomeric pump. The open amputation site was covered with adherent drapes and a negative-pressure wound therapy device provided vacuum-assisted closure (V.A.C.) for 72 hr. Three months later this animal developed a deep dermal ulcer on the lateral aspect of the right hind limb, at the level of the stifle. Methicillin-resistant Staphylococcus aureus was isolated. The wound was managed by initial daily lavage, followed by 1 mo of V.A.C. therapy, with 72 hr between dressing changes. Clinically, this therapy expedited the formation of healthy granulation tissue and overall healing was accelerated. The animal tolerated the machine and bandage changes well via operant conditioning. The use of negative-pressure wound therapy appeared to shorten time to resolution of slow-healing wounds in black rhinoceros.

ß-glucan from barley and its lipid-lowering capacity: a meta-analysis of randomized, controlled trials.

BACKGROUND/OBJECTIVES: To more precisely quantify the effect of barley ß-glucan on blood lipid concentrations in humans and to examine the factors that could affect its efficacy. SUBJECTS/METHODS: Eleven eligible randomized clinical trials published from 1989 to 2008 were identified from nine databases. Weighted mean effect sizes were calculated for net differences in lipid profile using a random effect model (RevMan 4.2). RESULTS: Overall, barley and ß-glucan isolated from barley lowered total and low-density lipoprotein (LDL) cholesterol concentrations by 0.30 mmol/l (95% confidence interval (CI): -0.39 to -0.21, P<0.00001) and 0.27 mmol/l (95% CI: -0.34 to -0.20, P<0.00001), respectively, compared with control. The pattern of cholesterol-lowering action of barley in this analysis could not be viewed as a dose-dependent response. There were no significant subgroup differences by type of intervention and food matrix. CONCLUSIONS: Increased consumption of barely products should be considered as a dietary approach to reduce LDL cholesterol concentrations.

The association between hip and groin injuries in the elite junior football years and injuries sustained during elite senior competition.

OBJECTIVE: To establish the relationship between the history of hip and groin injuries in elite junior football players prior to elite club recruitment and the incidence of hip and groin injuries during their elite career. DESIGN: Retrospective cohort study. SETTING: Analysis of existing data. PARTICIPANTS: 500 Australian Football League (AFL) players drafted from 1999 to 2006 with complete draft medical assessment data. ASSESSMENT OF RISK FACTORS: Previous history of hip/groin injury, anthropometric and demographic information. MAIN OUTCOME MEASUREMENT: The number of hip/groin injuries resulting in > or =1 missed AFL game. RESULTS: Data for 500 players were available for analysis. 86 (17%) players reported a hip/groin injury in their junior football years. 159 (32%) players sustained a hip/groin injury in the AFL. Players who reported a previous hip or groin injury at the draft medical assessment demonstrated a rate of hip/groin injury in the AFL >6 times higher (IRR 6.24, 95% CI 4.43 to 8.77) than players without a pre-AFL hip or groin injury history. CONCLUSIONS: This study demonstrated that a hip or groin injury sustained during junior football years is a significant predictor of missed game time at the elite level due to hip/groin injury. The elite junior football period should be targeted for research to investigate and identify modifiable risk factors for the development of hip/groin injuries.

Mapping quantitative trait loci controlling agronomic traits in the spring wheat cross RL4452x'AC Domain'.

Relatively little is known about the genetic control of agronomic traits in common wheat (Triticum aestivum L.) compared with traits that follow Mendelian segregation patterns. A doubled-haploid population was generated from the cross RL4452x'AC Domain' to study the inheritance of the agronomic traits: plant height, time to maturity, lodging, grain yield, test weight, and 1000-grain weight. This cross includes the genetics of 2 western Canadian wheat marketing classes. Composite interval mapping was conducted with a microsatellite linkage map, incorporating 369 loci, and phenotypic data from multiple Manitoba environments. The plant height quantitative trait loci (QTLs), QHt.crc-4B and QHt.crc-4D, mapped to the expected locations of Rht-B1 and Rht-D1. These QTLs were responsible for most of the variation in plant height and were associated with other agronomic traits. An additional 25 agronomic QTLs were detected in the RL4452x'AC Domain' population beyond those associated with QHt.crc-4B and QHt.crc-4D. 'AC Domain' contributed 4 alleles for early maturity, including a major time to maturity QTL on 7D. RL4452 contributed 2 major alleles for increased grain yield at QYld.crc-2B and QYld.crc-4A, which are potential targets for marker-assisted selection. A key test weight QTL was detected on 3B and prominent 1000-grain weight QTLs were identified on 3D and 4A.

Dietary protein concentration affects plasma arteriovenous difference of amino acids across the porcine mammary gland.

The objective of this study was to determine whether the porcine mammary gland responds to increasing dietary CP concentration through changes in AA arteriovenous difference (a-v). Sixteen Landrace x Yorkshire lactating sows were provided ad libitum access to one of four isocaloric diets varying in CP concentration (7.8, 13.0, 18.2, and 23.5 %; as-fed basis). Litters were adjusted to 11 pigs within 48 h of birth. Sows were fitted with catheters in the carotid artery and main mammary vein on d 4. On d 10, 14, 18, and 22 of lactation, arterial and venous blood samples were obtained every 30 min over 6 h. Milk yield was estimated on d 11 and 21 using the D2O dilution technique. Final litter sizes on d 21 were 10.3, 11, 9.5, and 11 piglets for sows fed the 7.8, 13.0, 18.2, and 23.5% CP diets, respectively. Piglet ADG tended (P = 0.088) to increase with increasing dietary CP concentration and were 186, 221, 220, and 202 g for sows fed the 7.8, 13.0, 18.2, and 23.5% CP diet, respectively. Daily total milk yield on d 21 (kg milk/d) tended (P = 0.099) to increase, and average milk yield per nursed piglet (kg of milk-pig(-1)d(-1)) increased (P < 0.05) with increasing CP concentration and were, on a per-piglet basis, 0.95, 1.19, 1.14 and 1.13 kg of milk/d for the 7.8, 13.0, 18.2, and 23.5% CP diets, respectively. As dietary CP increased from 7.8 to 23.5%, isoleucine and leucine a-v increased linearly only (linear, P < 0.01); all other AA a-v increased, reached a maximum in sows fed 18.2% CP, and decreased thereafter in sows fed 23.5% CP (quadratic, from P = 0.10 to P < 0.05). Amino acid uptake by the entire udder and by each gland increased (linear, P < 0.05) with increasing dietary CP. Arteriovenous differences response to increasing day of lactation varied among AA, from no change for histidine, isoleucine, lysine, methionine, tryptophan, and valine, to a linear trend increase for arginine (P = 0.055), leucine (P = 0.064), phenylalanine (P = 0.101), and threonine (P = 0.057). In summary, for the majority of AA, a-v increased with increasing dietary CP concentration from 7.8 to 18.2%, but decreased when CP concentration exceeded 18.2%. In contrast, mammary AA uptake, piglet ADG and milk yield per pig increased linearly with increasing dietary CP, suggesting a coordinated regulation between AA delivery and transport to meet the demand for milk yield.

Nitrogen balance and ileal amino acid digestibility in growing pigs fed diets reduced in protein concentration.

Two studies were conducted to assess the effect of dietary protein reduction on N utilization, N excretion, and AA digestibility in growing pigs. The objective was to determine whether pigs fed diets with a reduced CP concentration could maintain the same N retention as pigs fed an adequate diet. The second objective was to test whether reducing dietary CP concentration decreases AA digestibility. In each study, six barrows were allotted to one of six dietary treatments in a Latin square design. Treatments consisted of four corn-soybean meal-based diets containing 15, 12, 9, and 6% CP, a casein-based diet containing 15% CP, and a protein-free diet. Crystalline AA were included in the 12, 9, and 6% CP diets. The indispensable:dispensable AA ratio was maintained at 45:55 with the addition of L-glutamic acid to the 9 and 6% CP diets. The casein-based and protein-free diets were used to determine endogenous total tract N and ileal AA losses. In the first study, total N losses and N absorbed decreased linearly (P < 0.001) as dietary CP concentration decreased from 15 to 6%. Both a linear (P < 0.001) and a quadratic (P < 0.05) decrease in N retention were found with decreasing dietary CP concentration. Nitrogen retained as a percentage of intake and absorbed increased (P < 0.001) as dietary CP concentration was reduced from 15 to 6%. In the second study, six barrows were surgically fitted with a T-cannula at the terminal ileum to determine ileal AA digestibility. For all dispensable and most indispensable AA, apparent and standardized ileal digestibility increased linearly (P 0.01, and for arginine, P < 0.05) as dietary CP concentration decreased. These results indicate that dietary CP concentration can be decreased from 15 to 12% with crystalline AA supplementation to meet an ideal AA profile without adversely affecting N retention, and that decreasing dietary CP concentration from 15 to 6% increases both dispensable and indispensable AA ileal digestibility.

Characterization of the predominant anaerobic bacterium recovered from digital dermatitis lesions in three Michigan dairy cows.

Digital dermatitis is a superficial epidermatitis of the feet of cattle. Data from previous work suggest that spirochaetes, Campylobacter spp., and Bacteroides spp. may be important in the disease, but the etiology of this disease is not entirely clear. Tissue samples collected from digital dermatitis lesions in three Holstein-Friesian cows from a Michigan dairy yielded a predominant colony type when incubated anaerobically on blood agar at 35 degrees C for 24-48 h. The isolate was a non-flagellated Gram-negative rod, 7 microM long and <0.5 microM wide; its growth was strictly anaerobic and resulted in slight ss-hemolysis on blood agar; 16S rRNA gene sequence analysis indicated it belonged to the cytophoga-flexibacter-bacteroides phylum. The finding that this bacterium was the predominant anaerobe recovered from digital dermatitis lesions suggests it may be involved in the digital dermatitis disease process.

Abnormal brain activation on functional MRI in cognitively asymptomatic HIV patients.

BACKGROUND/OBJECTIVES: A previous fMRI study demonstrated increased brain activation during working memory tasks in patients with HIV with mild dementia. The current study aims to determine whether patients who are HIV-1 positive and have normal cognitive function also show increased brain activation on fMRI. METHODS: Blood oxygenation level-dependent (BOLD) fMRI was performed in 10 patients with HIV (CD <500) and 10 age-, sex-, education-, and handedness-matched seronegative subjects. Each subject performed a battery of neuropsychological tests and fMRI with three tasks (0-back, 1-back, and 2-back) that required different levels of attention for working memory. RESULTS: Compared with control subjects, patients with HIV showed greater magnitude of brain activation (BOLD signal intensity changes, p

Neutral detergent fiber concentration of corn silage and rumen inert bulk influences dry matter intake and ruminal digesta kinetics of growing steers.

Corn silage with high NDF concentration has the potential to reduce DMI because it has a greater filling effect in the rumen than low-NDF corn silage. Our objective was to determine whether ruminal fill influences DMI to the same extent with low- or high-NDF corn silage-based diets. Eight ruminally cannulated Holstein steers (198 +/- 13 kg) were randomly assigned to a 2 x 2 factorial arrangement of treatments in a replicated 4 x 4 Latin square design with 16-d periods. Treatments were diets containing corn silage from a normal hybrid (low-fiber; LF) or its male-sterile isogenic counterpart (high-fiber; HF), offered for ad libitum consumption to steers with or without rumen inert bulk (RIB). The LF and HF diets contained 33.8 and 50.8% dietary NDF, respectively. Rumen inert bulk was added at 25% of pretrial ruminal volume in the form of plastic-coated tennis balls filled with sand to achieve a specific gravity of 1.1 and a total volume of 7.5 L. No fiber level x inert bulk interactions were detected for DMI or NDF intake (P > 0.10), suggesting that DMI was limited to the same extent by physical fill at both levels of dietary fiber. Addition of RIB decreased DMI by an average of 10.7%, which was 65.5 g/L of added bulk. The HF diet depressed DMI by an average of 15.5%, increased NDF intake 27.1%, and reduced ruminal NDF turnover time by 21.0% compared to the LF diet (P < 0.01), with no effect on ruminal volume or amount of NDF in the rumen (P > 0.10). Addition of RIB also reduced ruminal NDF turnover time and amount of NDF in the rumen (11.8% and 20.7%, respectively; P < 0.01), with no change in ruminal digesta volume (P > 0.10). The HF treatment decreased digestibility of DM and GE (5.5 and 5.7%, respectively; P < 0.01) but increased NDF digestibility (10.4%; P < 0.01) compared to LF. Rumen inert bulk had no effect on digestibility of DM, NDF, or GE (P > 0.10). The lack of reduction in digesta volume with addition of inert fill suggests that DMI of light-weight steers receiving corn silage-based diets within a wide range of NDF concentrations was not regulated by ruminal distension alone.

In utero hematopoietic stem cell transplantation: a caprine model for prenatal therapy in inherited metabolic diseases.

OBJECTIVES: We explored the feasibility and efficacy of in utero hematopoietic stem cell transplantation in the caprine animal model system with the objectives of determining procedures for transplantation and establishing methods for detecting engraftment. METHODS: Male fetal liver hematopoietic stem cells were injected into female fetuses during the immunotolerant period, using either hysterotomy or ultrasound-guided injections. RESULTS: The rate of fetal death was much lower for the ultrasound-guided injections. Donor cells were observed in the peritoneal fluid of 4 fetuses 3 days after injection, but no donor cells were detected in tissues at longer time periods. CONCLUSIONS: Ultrasound-guided injection of hematopoietic stem cells into the abdomen of a developing fetus is safe and feasible. The parameters required for successful engraftment have not yet been identified.

Brown midrib-3 corn silage improves digestion but not performance of growing beef steers.

The brown midrib-3 (bm3) gene mutation has been incorporated into corn plants to potentially improve fiber digestibility. The objectives of this study were to determine the effect of bm3 corn silage on digestion and performance of growing beef steers and to determine whether limiting intake would further enhance fiber digestibility of bm3 corn silage. A bm3 hybrid and its isogeneic normal counterpart were harvested at three-quarters kernel milk line. Neutral detergent fiber, ADF, and ADL were 4.5, 6.9, and 1.9 units lower, respectively, and DM was 5.4 units higher for bm3 than for normal silage. In Trial 1, eight ruminally fistulated Angus crossbred steers (224 +/- 24 kg) were randomly assigned to a 2 x 2 factorial arrangement of treatments in a replicated 4 x 4 Latin square design. Steers had ad libitum feed access or were restricted to 80% of ad libitum intake of diets containing 86% normal corn silage (Control) or bm3 corn silage (BMCS). The remainder of the diets consisted of soybean meal, urea, monensin, vitamins, and minerals. Dry matter intake was greater (P < 0.01) for steers offered ad libitum access to BMCS than for those with ad libitum access to the Control diet. The BMCS treatment resulted in improved (P < 0.05) apparent total-tract digestibility of DM, OM, NDF, and ADF. Mean concentration of total VFA and molar proportions of acetate were increased (P < 0.05) by feeding BMCS. There tended to be a DMI x hybrid interaction (P = 0.16) for apparent total-tract digestibility of NDF. When diets were offered ad libitum, BMCS increased NDF digestibility by 10.5 percentage units compared with Control, but, when DMI was limited, BMCS increased NDF digestibility by 15.8 percentage units. In Trial 2, 128 steer contemporaries of those used in Trial 1 (245 +/- 13 kg) were offered ad libitum access to BMCS or Control diets as used in Trial 1. After a 112-d treatment period, concentrate in the diet was increased, and all steers were fed a common finishing diet. During the 112-d treatment period, steers receiving BMCS consumed 0.45 kg more DM/d (P < 0.05) and had similar ADG (P > 0.10), compared with those steers receiving the Control silage. This resulted in poorer (P < 0.01) feed efficiency for steers receiving BMCS. Finishing phase and overall performance of the steers was not different (P > 0.10) due to treatment. Although feeding BMCS in growth-phase diets resulted in increased daily DMI and improved digestibility of DM and fiber, it did not result in improved steer feedlot ADG compared with Control silage.

Grazing methods and stocking rates for direct-seeded alfalfa pastures: II. Pasture quality and diet selection.

A 2-yr study was conducted to determine the effects of two grazing methods (GM) and two stocking rates (SR) on alfalfa (Medicago sativa L. var. WL225) pasture quality and diet selection by Holstein steers. Eight pasture plots (.76 ha) were seeded in 1988 and divided into two blocks of four pastures each. Pastures were managed to allow a 36-d rest period with an average grazing season of 105 d. Before steers entered the next paddock, canopy heights (CH) of alfalfa plants were determined and pasture-forage samples were collected. Forage samples were analyzed for DM, OM, CP, and in vitro OM digestibility (IVOMD). At 12-d intervals beginning with the second grazing cycle, extrusa samples were collected from steers with esophageal fistulas. Extrusa samples were frozen, freeze-dried, and analyzed for OM, CP, IVOMD, in situ ruminal DM degradation, and ruminal undegradable protein. There were no effects of GM on alfalfa CH or pasture DM, OM, CP, and IVOMD. Increasing the SR increased pasture CP content in both years and increased DM, OM, and IVOMD in the 2nd yr. There was no effect of GM or SR on the quality of forage selected by esophageally fistulated steers. Esophageally fistulated steers selected forage that had greater OM, CP, and IVOMD than the average nutrient content of the forage. Although forage quality was greater when stocking rates were increased, the quantity of forage available per animal may have limited gains.

Diagnosis and management of a malignant granulosa cell tumor in a holstein nulligravida: a case study.

Transrectal palpation of an 18-mo-old, anestrous Holstein nulligravida revealed an enlarged left ovary. Ultrasonographically, the mass resembled known equine granulosa cell tumor (GCT) images. Slowly, virilism developed. Fifty-five days after initial diagnosis, unilateral ovariectomy was performed The presurgical serum concentration of testosterone was equal to that of a mature bull. During the 3 mo period following surgery, estrus expression resumed, the serum testosterone concentration returned to baseline, and the heifer was diagnosed pregnant following AI. A low grade, malignant GCT was the final histopathologically diagnosis. Clinical diagnosis of GCT should involve ultrasonography and steroid hormone assay. An ultrasound image, similar to the one shown, may be characteristic of GCT in the bovine.

Biochemical and morphological expression of early prenatal caprine beta-mannosidosis.

Lysosomal storage diseases associated with early-onset pathological changes may require prenatal therapy to avert the profound effects of the metabolic error on organs, especially the central nervous system. The present investigation determined the extent of expression of beta-mannosidase deficiency in the caprine fetus at 62 days of gestation, near the end of the period of immunotolerance when donor cells can engraft in various organs without immune rejection and supply missing enzyme. Three pairs of obligate carrier goats from the beta-mannosidosis colony were mated. Out of six fetuses delivered at 62 days of gestation, one (V385) was identified by measurement of beta-mannosidase activity as the only fetus affected with beta-mannosidosis. Thin-layer chromatography and quantitation of oligosaccharides revealed the presence of tri- and disaccharides, typical of beta-mannosidosis, only in V385. Morphological analysis revealed cytoplasmic vacuolation typical of beta-mannosidosis in V385; in thyroid, spinal cord, and kidney, the pattern of vacuolation was similar to, but less severe than, that observed previously in newborn affected goats. On the basis of these results, it will be possible to determine the effects of prenatal cell transplantation therapeutic strategies performed during the period of immunotolerance by monitoring phenotypic characteristics after treatment.

Development and efficacy of ultrasound-guided fetal fluid aspiration techniques for prenatal diagnosis of caprine beta-mannosidosis.

Ultrasound-guided fetal fluid sampling was performed on 13 pregnant goats at Days 59 to 65 of gestation to establish safe techniques for accurate sampling and to determine the feasibility of prenatal diagnosis of beta-mannosidosis. Fluids were analyzed for electrolyte and creatinine content to assess accuracy of sampling. Values correlated well with previously reported concentrations for caprine and ovine fetal fluids at the same gestational stage. The single abortion which occurred following ultrasound-guided sampling was correlated with placentome penetration and aspiration of bloody fluids. Thin layer chromatography of amniotic and allantoic fluids was performed to detect oligosaccharides that accumulate in beta-mannosidosis. Abnormal accumulated oligosaccharides were identified in the allantoic but not amniotic fluid from a beta-mannosidase-deficient 62-d-old fetus. Thus, allantocentesis was shown to be an optimal, safe procedure for providing information at this gestational stage to diagnose caprine beta-mannosidosis.

Use of a fracture distractor in two cattle.

Open reduction and internal fixation was attempted in a 16-month-old cow with a fracture of the femur and a 10-month-old heifer with a fracture of the humerus. In both animals, manual reduction of the fracture was unsuccessful, but following application of a fracture distractor, reduction was easily accomplished.

Reduction of bovine plasma cholesterol concentration by partial interruption of enterohepatic circulation of bile salts: a novel hypocholesterolemic model.

Interruption of enterohepatic circulation (EHC) of bile salts in several species is known to cause a significant decrease in plasma concentrations of low density lipoprotein (LDL) cholesterol, but to have little effect on high density lipoprotein (HDL) cholesterol. The present study, for the first time, demonstrates that partial interruption of EHC dramatically reduces both plasma LDL and HDL cholesterol concentrations in cattle. Five adult Holstein cows were surgically altered to allow controlled portions of bile flow to be diverted from the body. The animals were fed a low-fat, cholesterol-free diet. In two experiments, bile was diverted at 50% and 22% of total flow rates. By day 8 of diversion, both rates reduced mean plasma cholesterol from baseline (85 mg/dl) to about 8 and 18 mg/dl, respectively. Cholesterol was reduced in equal proportions in all lipoprotein fractions. In addition, plasma concentrations of triglycerides and phospholipids were also dramatically reduced. All of these plasma lipids returned to baseline within 1 week after restoration of bile flow. To determine the hepatic response to bile diversion, liver cholesterol concentrations, cholesterol synthesis rates, and LDL receptor-binding activities were determined in biopsy samples. In response to bile diversion, hepatic cholesteryl esters were markedly depleted while hepatic cholesterol synthesis rates were increased by more than 10-fold. Nevertheless, because the basal cholesterol synthesis rate was so low, it was estimated that the increase in synthesis would have supplied no more than 5% of the sterols depleted during bile diversion (1.2 vs. 25 mmol/day). LDL receptor-binding activity was significantly elevated, suggesting an increased uptake of plasma lipoprotein cholesterol by the liver. These results suggest that the unique sensitivity of bovine plasma cholesterol to enterohepatic circulation interruption might occur as a result of the inherently low rate of hepatic cholesterol synthesis in cattle. This hypocholesterolemic model might serve as an interesting tool for the study of factors regulating plasma HDL cholesterol.