Effects of reducing dietary ([Na + K − [Cl + SO4]) on bone in dairy cows at parturition

The effects of feeding diets with different milliequivalents (mEq) of dietary ([Na⁺ + K⁺] − [Cl⁻ + SO₄⁼]) to dairy cows during the last seven weeks of pregnancy on bone morphology at parturition were studied. Nine monozygotic twin pairs of pregnant cows (five pairs of parity 1 or 2 and four pairs of parity 3 or more) were allocated to two diets which were formulated to provide either −4 mEq (anion diet) or +572·5 mEq (cation diet) of ([Na⁺ + K⁺] − [Cl⁻ + SO₄⁼]) kg⁻¹ dietary dry matter. Bone biopsies were taken from the tuber coxae between three and eight hours after parturition. The plasma concentrations of calcium and inorganic phosphorus, the total plasma alkaline phosphatase activity and the urinary hydroxyproline:creatinine ratio were not significantly affected by diet during the experimental period. In low parity (2 or less) cows the percentage trabecular bone volume, the percentage osteoclast surface and the mean number of osteoclasts per microscopic field (identified by Goldner staining) were lower on the anion diet than on the cation diet (P<0-02). In the high parity cows, the percentage osteoid volume (P<0·05) and the ratio of percentage osteoid volume to percentage osteoid surface (P<0·001) were greater in the cows fed the anion diet than in the cows fed the cation diet. The results show that reducing the mEq of dietary ([Na⁺ + K⁺] − [Cl⁻ + SO₄⁼]) to −4 mEq kg⁻¹ dietary dry matter affected some of the parameters of bone formation but did not enhance bone resorption.     

Characterization of Equine Phospholipase C Zeta: A Review and Preliminary Results on Expression Defects in Subfertile Stallions

In all mammalian species studied thus far, fertilization results in a series of intracellular Ca²⁺ ([Ca²⁺]_i) increases, referred to as oscillations, responsible for driving oocyte activation and embryonic development. Current evidence supports the notion that sperm-borne phospholipase C zeta (PLCZ) is responsible for the initiation of these [Ca²⁺]_i oscillations. Although this appears to be a highly conserved mechanism for oocyte activation, differences in PLCZ sequence, activity, and expression do exist among different species. Herein, we summarize the information supporting PLCZ as the oocyte-activating factor in mammals and present our current knowledge regarding the characterization of this protein in the horse. The equine sequence yielded a protein of high relative [Ca²⁺]_i-releasing activity. Equine PLCZ was expressed over the head region overlying the acrosome, equatorial segment, connecting piece between the head and midpiece, and on the principal piece of the flagellum of stallion sperm. Equine PLCZ expressed both over the head and tail sperm regions was catalytically active, with the latter representing a characteristic unique to the horse. We also present preliminary data in subfertile stallions displaying PLCZ expression defects, although further research is required to establish a clear association between these defects and fertility problems in the horse. In summary, the information presented raises the questions of whether equine PLCZ could play diverse roles in sperm physiology and/or become a marker for the evaluation of stallion fertility, both of which are worthy of further investigation.     

Inositol-1,4,5-Trisphosphate Receptor-1 and -3 and Ryanodine Receptor-3 May Increase Ooplasmic Ca2+ During Quail Egg Activation

We previously reported that egg activation in Japanese quail is driven by two distinct types of intracellular Ca²⁺ ([Ca²⁺]i): transient elevations in [Ca²⁺]i induced by phospholipase Czeta 1 (PLCZ1) and long-lasting spiral-like Ca²⁺ oscillations by citrate synthase (CS) and aconitate hydratase 2 (ACO2). Although the blockade of inositol 1,4,5-trisphosphate receptors (ITPRs) before microinjections of PLCZ1, CS, and ACO2 cRNAs only prevented transient increases in [Ca²⁺]i, a microinjection of an agonist of ryanodine receptors (RYRs) induced spiral-like Ca²⁺ oscillations, indicating the involvement of both ITPRs and RYRs in these events. In this study, we investigated the isoforms of ITPRs and RYRs responsible for the expression of the two types of [Ca²⁺]i increases. RT-PCR and western blot analyses revealed that ITPR1, ITPR3, and RYR3 were expressed in ovulated eggs. These proteins were degraded 3 h after the microinjection of PLCZ1, CS, and ACO2 cRNAs, which is the time at which egg activation was complete. However, degradation of ITPR1 and ITPR3, but not RYR3, was initiated 30 min after a single injection of PLCZ1 cRNA, corresponding to the time of the initial Ca²⁺ wave termination. In contrast, RYR3 degradation was observed 3 h after the microinjection of CS and ACO2 cRNAs. These results indicate that ITPRs and RYR3 differentially mediate in creases in [Ca²⁺]i during egg activation in Japanese quail, and that downregulation of ITPRs and RYR3-mediated events terminate the initial Ca²⁺ wave and spiral-like Ca²⁺ oscillations, respectively.     

Mechanisms involved in the cytosolic calcium ion elevation induced by activating factors in chicken and rat phagocytes

The aim of the present study was to determine the mechanism of cytosolic calcium ion concentration [Ca²⁺]_i elevation in chicken and rat phagocytes stimulated with phorbol myristate acetate (PMA), leukotriene B₄ (LTB₄), formyl‐methionyl‐leucyl‐phenylananine (fMLP) and Saccaromyces cerevisiae culture supernatant (SCS). Pretreatment with EGTA completely suppressed the PMA‐induced [Ca²⁺]_i elevation in rat and chicken phagocytes, suggesting that all the [Ca²⁺]_i elevation induced in the PMA‐stimulated rat and chicken phagocytes was attributable to the influx of extracellular Ca²⁺. On the other hand, the elevation of LTB₄‐, FMLP‐ and SCS‐induced [Ca²⁺]_i was only partially suppressed by ethyleneglycol‐bis (β‐aminoethyl)‐N,N,N′,N′‐tetraacetic acid ethylene (EGTA) pretreatment of phagocytes. The results indicated that two pathways of [Ca²⁺]_i elevation, recruitment from the intracellular Ca²⁺ store and influx of extracellular Ca²⁺, are involved in the [Ca²⁺]_i elevation of LTB₄‐, fMLP‐ and SCS‐stimulated phagocytes. In fMLP‐stimulated rat neutrophils, [Ca²⁺]_i elevation showed a two‐phase pattern in which the time lag between the first and second phase was approximately 1 min. The EGTA treatment of the fMLP‐stimulated cells induced a reduction of the first phase level and a disappearance of the second phase. The reason for the special influence of EGTA observed in fMLP‐stimulated cells is unknown, but the disappearance of the second phase of the [Ca²⁺]_i may be elicited by the EGTA‐induced decrease of the first phase [Ca²⁺]_i elevation that depends on IP₃ and diacylglycerol induced by fMLP.     

Growth hormone release induced by an amino acid mixture from primary cultured anterior pituitary cells of goats

The effects of amino acids on growth hormone (GH) release and cytosolic calcium concentration ([Ca²⁺]_i) were investigated in caprine anterior pituitary cells cultured for 3 d in Dulbecco modified Eagle medium. The addition of an amino acid mixture consisting of seven nonessential amino acids (NEAA: l-Asp, Gly, l-Ala, l-Ser, l-Pro, l-Asn, and l-Glu; concentration of each 12.5–200 μmol/l) in the medium significantly raised GH release from the cultured cells in a concentration-dependent manner with the maximum release at 200 μmol/l NEAA. Although an addition of l-Asp (0.1–100 μmol/l) caused a significant rise in GH release in a concentration-dependent manner, neither the individual amino acids contained in NEAA except l-Asp nor others (l-Leu, l-Phe, l-Gln, l-Met, and l-Arg) caused a rise in GH release when added alone to the medium. The rise in GH release induced by NEAA (200 μmol/l) and GH-releasing hormone (GHRH, 10 nmol/l) was significantly reduced by the addition of EGTA (l.8 mmol/l) and nifedipine (1 μmol/l) to the medium, respectively. The addition of NEAA (200 μmol/l) caused a rapid and transient [Ca²⁺]_i increase, followed thereafter by a steady increase. The prior addition of nifedipine (1 μmol/l), which itself significantly reduced the basal [Ca²⁺]_i, completely abolished the response induced by NEAA or GHRH. From these findings, we conclude that: 1) NEAA raises GH release and [Ca²⁺]_i in cultured caprine anterior pituitary cells, and 2) Ca²⁺ influx from the medium may be responsible for the cellular action of NEAA.     

The addition of calcium ion chelator,EGTA to thawing solution improves fertilizing ability in frozen–thawed boar sperm

In our previous study, seminal plasma effectively suppressed the induction of sperm to capacitation‐like status and acrosome loss during the thawing process. However, because boar seminal plasma is contaminated with various kinds of bacteria and/or viruses, it is necessary to develop a thawing solution without animal‐derived materials. In this study, we focused on the increase of intracellular Ca²⁺ ([Ca²⁺]_i) in sperm after thawing and the negative effects of sperm qualities. After thawing, the fluorescent intensity of [Ca²⁺]_i indicator, Fluo‐3/AM, and the level of phosphorylated tyrosine residue of protein were increased in the sperm. Next, we investigated whether the addition of Ca²⁺ chelators (ethylenediaminetetraacetic acid (EDTA) and ethyleneglycoltetraacetic acid (EGTA)) improved post‐thawed sperm motility. When the frozen–thawed sperm were treated with 6 mmol/L EDTA + 6 mmol/L EGTA, sperm motility was significantly increased as compared with control (6 mmol/L EDTA alone) at all incubation periods (P < 0.05). The combinational treatment significantly suppressed the elevation of [Ca²⁺]_i and the tyrosine phosphorylation, which improved the acrosomal status and fertilizing ability in vitro. Furthermore, when the thawing method was applied for artificial insemination, the fertilization rate was significantly higher than control (P < 0.05, 33% vs. 82%). Thus, we conclude that the addition of EDTA + EGTA to thawing solution is a beneficial tool for artificial insemination using frozen–thawed boar sperm.     

Involvement of the Ca2+ signaling pathway in osteoprotegerin inhibition of osteoclast differentiation and maturation

The purpose of this study was to determine whether the Ca²⁺ signaling pathway is involved in the ability of osteoprotegerin (OPG) to inhibit osteoclast differentiation and maturation. RAW264.7 cells were incubated with macrophage colony-stimulating factor (M-CSF) + receptor activator of nuclear factor-κB ligand (RANKL) to stimulate osteoclastogenesis and then treated with different concentrations of OPG, an inhibitor of osteoclast differentiation. The intracellular Ca²⁺ concentration [Ca²⁺]_i and phosphorylation of Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) in the different treatment groups were measured by flow cytometry and Western blotting, respectively. The results confirmed that M-CSF + RANKL significantly increased [Ca²⁺]_i and CaMKII phosphorylation in osteoclasts (p < 0.01), and that these effects were subsequently decreased by OPG treatment. Exposure to specific inhibitors of the Ca²⁺ signaling pathway revealed that these changes varied between the different OPG treatment groups. Findings from the present study indicated that the Ca²⁺ signaling pathway is involved in both the regulation of osteoclastogenesis as well as inhibition of osteoclast differentiation and activation by OPG.     

The concentration of ionized magnesium in serum during the periparturient period of non-paretic dairy cows

Ion-selective electrodes have recently been designed for determining the ionized concentration of magnesium (Mg²⁺) in serum. This development may allow new insights into some metabolic diseases of cattle. For this report, the concentrations of Mg²⁺, total magnesium (Mg_tot), ionized calcium (Ca²⁺), total calcium (Ca_tot), and inorganic phosphate (P_i) were determined in sera from seventeen 3-to 16-year-old Brown Swiss and crossed Simmental/Red Holstein cows during the periparturient period. In each animal, a transient increase of Mg²⁺ and Mg_tot serum concentrations was observed in association with the transient decrease in serum concentrations of Ca²⁺, Ca_tot and P_i after parturition. On average, throughout the study, the serum Mg²⁺ concentrations were 68.5% of those of Mg_tot, whereas the serum Ca²⁺ concentrations were 52% of those of Ca_tot. The possible mechanisms involved in the transient increase of Mg²⁺ and Mg_tot serum concentrations are discussed.Abbreviations Ca²⁺ ionized calcium - Ca_tot total calcium - Mg²⁺ ionized magnesium - Mg_tot total magnesium - P_i inorganic phosphate - PTH parathyroid hormone - PTHrP parathyroid homrone related protein     

Physicochemical determinants of pH in pectoralis major of three strains of laying hens housed in conventional and furnished cages

1. Post-mortem decline in muscle pH has traditionally been attributed to glycogenolysis-induced lactate accumulation. However, muscle pH ([H⁺]) is controlled by complex physicochemical relationships encapsulated in the Stewart model of acid–base chemistry and is determined by three system-independent variables – strong ion difference ([SID]), total concentration of weak acids ([A_tot]) and partial pressure of CO₂ (PCO₂).

2. This study investigated these system-independent variables in post-mortem pectoralis major muscles of Shaver White, Lohmann Lite and Lohmann Brown laying hens housed in conventional cages (CC) or furnished cages (FC) and evaluated the model by comparing calculated [H⁺] with previously measured [H⁺] values.

3. The model accounted for 99.7% of the variation in muscle [H⁺]. Differences in [SID] accounted for most or all of the variations in [H⁺] between strains. Greater PCO₂ in FC was counteracted by greater sequestration of strong base cations. The results demonstrate the accuracy and utility of the Stewart model for investigating determinants of meat [H⁺].

4. The housing differences identified in this study suggested that hens housed in FC have improved muscle function and overall health due to the increased opportunity for movement. These findings support past studies showing improved animal welfare for hens housed in FC compared to CC. Therefore, the Stewart model has been identified as an accurate method to assess changes in the muscle at a cellular level that affect meat quality that also detect differences in the welfare status of the research subjects.     

Indication of intracellular magnesium deficiency in lactating dairy cows revealed by magnesium loading and renal fractional excretion

Nine non‐pregnant, lactating dairy cows were used to study plasma and urinary magnesium concentrations ([Mg]_pl; [Mg]_u), and the urinary fractional excretion of magnesium (FE_Mg) before, during and after an 120 min intravenous magnesium (Mg) administration (2.5 mg/kg body weight). Animals received a total mixed ration, and Mg content of the diet was within recommended range. Basal mean [Mg]_pl, [Mg]_u and FE_Mg were 0.89 ± 0.09 mm , 5.92 ± 2.99 mm and 8.3 ± 9.7% respectively. For all parameters, a substantial inter‐individual variation was observed. Three cows showed suboptimal [Mg]_pl and/or [Mg]_u as well as low FE_Mg values of approximately 2% indicating an insufficient Mg supply to these animals (depressed feed intake, reduced absorption of Mg). The applied Mg challenge induced no significant change of mean [Mg]_pl in the cows because part of the excess Mg was excreted in the urine. But in five out of nine cows, a decrease of the FE_Mg, during and after an intravenous Mg load was observed showing that part of the infused Mg is used to replenish intracellular Mg pools. Thus, the existence of an intracellular Mg deficiency in these cows was unmasked by performing the Mg loading test only. Because a reduced free intracellular [Mg] impairs cell and tissue functions, the results highlight the importance of an accurate definition of the intracellular Mg status. The Mg loading test is a suitable procedure, however, for practical purposes less expensive and time consuming methods must be developed.     

Effect of Intravenously Administered Crystalloid Solutions on Acid‐Base Balance in Domestic Animals

Intravenous fluid therapy can alter plasma acid‐base balance. The Stewart approach to acid‐base balance is uniquely suited to identify and quantify the effects of the cationic and anionic constituents of crystalloid solutions on plasma pH. The plasma strong ion difference (SID) and weak acid concentrations are similar to those of the administered fluid, more so at higher administration rates and with larger volumes. A crystalloid's in vivo effects on plasma pH are described by 3 general rules: SID > [] increases plasma pH (alkalosis); SID < [] decreases plasma pH (alkalosis); and SID = [] yields no change in plasma pH. The in vitro pH of commercially prepared crystalloid solutions has little to no effect on plasma pH because of their low titratable acidity. Appreciation of IV fluid composition and an understanding of basic physicochemical principles provide therapeutically valuable insights about how and why fluid therapy can produce and correct alterations of plasma acid‐base equilibrium. The ideal balanced crystalloid should (1) contain species‐specific concentrations of key electrolytes (Na⁺, Cl^?, K⁺, Ca⁺⁺, Mg⁺⁺), particularly Na⁺ and Cl^?; (2) maintain or normalize acid‐base balance (provide an appropriate SID); and (3) be isosmotic and isotonic (not induce inappropriate fluid shifts) with normal plasma.     

Molecular characteristics of horse phospholipase C zeta (PLCζ)

A sperm‐specific phospholipase C (PLC), PLCzeta (PLCζ), is thought to underlie the initiation of calcium ([Ca²⁺]_i) oscillations that induce egg activation in mammals. In large domestic species, only bovine, porcine and recently equine PLCζ have been cloned, and the physiological functions of these molecules have not been fully characterized. Here, we evaluated the physiological functions of equine PLCζ (ePLCζ) in mouse oocytes. ePLCζ was cloned from testis using RT‐PCR. The expression of ePLCζ messenger RNA was confirmed in testis but not in other tissues. Microinjection of ePLCζ complementary RNA (cRNA) into mouse oocytes induced long‐lasting [Ca²⁺]_i oscillations, and most of the injected oocytes formed pronuclei (PN). The injection of cRNAs encoding horse, mouse, human and cow PLCζ into mouse oocytes showed that ePLCζ had the highest [Ca²⁺]_i oscillation‐inducing activity among the species tested. Mutation of D202R, which renders the protein inactive, abrogated the activity of ePLCζ. The nuclear translocation ability of ePLCζ was defective when expressed in mouse oocytes. Taken together, our findings show for the first time that ePLCζ has highest activity of the mammalian species studied to date. Our findings will be useful for the improvement of reproductive technologies in the horse.     

Physiological Changes in the Red Drum after Long-Term Freshwater Acclimation

Abstract

The effect of long-term freshwater acclimation on the blood and plasma ion composition of Red Drum Sciaenops ocellatus was investigated with the goal of elucidating the necessity of ion remediation. Four replicates (n = 50) of freshwater-acclimated (FW) fish (1.6 ± 0.2 g) were raised in 25-m³ tanks supported by 140,000 L of recirculating water. Four replicates (n = 50) of seawater (SW) fish groups were placed in 40-m³ offshore cages at 32–35 psu. Blood was collected from 100 fish (FW = 578 ± 50 g; SW = 686 ± 45 g) of each group (FW, SW) after 8 months of rearing. During the grow-out phase, the survival of FW and SW fish was 57.5% and 92.2%, respectively. The water ion composition (mainly the Ca²⁺/K⁺ [43%] and Ca²⁺/Mg²⁺ ratios [1%]) explained 56.6% of the plasmatic ion variability in the fish groups. Freshwater exposure produced significant reductions in osmolality and in several plasma indicators (Na⁺, Cl^?, and Mg²⁺); the K⁺ levels from FW fish were the most compromised parameter. The water Ca²⁺/Na⁺ ratio had a greater influence (44%) on the plasma chemistry parameters, mainly glucose and creatinine. Freshwater-acclimated fish had a higher percentage of hematocrit, hemoglobin, and red blood cells than SW fish, but the water quality explained only 12.5% of the blood parameter variability between the FW and SW groups. The results support the conclusion that Red Drum tolerates salinity variations and can adopt a relatively stable condition for short periods; however, the data suggest that Red Drum have only a limited ability to withstand a hyposmotic environment for long periods due to their limited ability in maintaining K⁺ concentrations without external supplementation. Freshwater environments with high Ca²⁺/Na⁺, Ca²⁺/K⁺, and Ca²⁺/Mg²⁺ ratios appear to be a chronic stress factor that should be considered in future experiments.

Received June 11, 2012; accepted March 17, 2013     

Spermatozoa Concentration,Seminal Plasma Composition and Their Physiological Relationship in the Endangered Stellate Sturgeon (Acipenser stellatus) and Russian Sturgeon (Acipenser gueldenstaedtii )

Comparative studies of ionic composition, osmolality, protein concentration and pH of seminal plasma along with spermatozoa concentrations were carried out in stellate sturgeon, Acipenser stellatus, and Russian sturgeon, Acipenser gueldenstaedtii. Analysis of A. gueldenstaedtii sperm showed significantly higher concentrations of Na⁺ (34.58 ± 4.61 mm ), Ca²⁺ (0.35 ± 0.12 mm ), Mg²⁺ (0.70 ± 0.25 mm ), Cl^? (13.50 ± 4.04 mm ) and proteins (0.60 ± 0.29 mg/ml) in the seminal plasma than did seminal plasma of A. stellatus: Na⁺ (20.08 ± 10.75 mm ), Ca²⁺ (0.28 ± 0.06 mm ), Mg²⁺ (0.29 ± 0.05 mm ), Cl^? (7.50 ± 3.00 mm ) and 0.30 ± 0.11 mg/ml proteins. Significantly higher concentration of K⁺ (5.42 ± 1.06 mm ) was observed in A. stellatus compared to A. gueldenstaedtii K⁺ (2.29 ± 0.50 mm ). Concentration of Na⁺ was positively correlated with osmolality (r = 0.819), levels of Cl^? (r = 0.922) and Mg²⁺ (r = 0.727) and pH (r = 0.848). The concentration of Mg²⁺ was positively correlated with protein concentration (r = 0.774), Na⁺ (r = 0.727), Cl^? (r = 0.872) and Ca²⁺ (r = 0.801). A positive relationship was also found between concentration of K⁺ and spermatozoa concentration (r = 0.709). Results revealed strong inter‐species differences in several parameters. The data should be useful for artificial fertilization and for cryopreservation of sturgeon sperm.     

Calcium homeostasis and its relationship to superoxide production in blood and milk neutrophils of lactating goats

Polymorphonuclear neutrophils (PMN), which comprise over 70% of the somatic cells in goat milk, are a major cellular component of innate immunity in the goat mammary gland. However, the function of milk PMNs is modified after diapedesis compared to PMNs in blood. As many aspects of PMN activity depend directly on intracellular Ca²⁺ concentration ((Ca²⁺)_i), the present study aimed to determine the changes in Ca²⁺ homeostasis of milk PMNs from lactating goats compared to autologous blood PMNs, and to examine the significance of these variations to the immuno-competency of milk PMNs. The intracellular Ca²⁺ store of freshly prepared milk cells was estimated from the elevation of (Ca²⁺)_i after ionomycin treatment, which was found to be significantly less than blood PMNs. Replenishment of the intracellular Ca²⁺ store in milk cells after intracellular Ca²⁺ depletion by Bapta-AM followed by spiking with 2.5 mM Ca²⁺ for 20 min was also compared to that of blood PMNs, showing that after depletion/spiking the intracellular Ca²⁺ store in milk cells was much less than blood PMNs. The production of superoxide anion (O₂^?) in vitro in response to (Ca²⁺)_i-dependent or (Ca²⁺)_i-independent modulators was used to evaluate the relevance of altered Ca²⁺ homeostasis on the immuno-competency of milk cells compared to blood PMNs. The results indicated that milk cells produced similarly low levels of O₂^? as blood PMNs when treated with ionomycin. However, the amount of O₂^? produced by milk cells in response to phorbol 12-myristate 13-acetate (PMA) stimulation, although greater than ionomycin treatment, was significantly less than that of blood PMNs. The capacity for O₂^? production by both cell types in response to PMA reverted to the resting state with use of the protein kinase C (PKC) inhibitor, staurosporine. In conclusion, the current study demonstrated an irreversible shortage of intracellular Ca²⁺ in the milk PMNs of lactating goats compared to blood PMNs. It also showed that preliminary O₂^–production, primed by ionomycin treatment, remained unchanged in milk PMNs, despite the shortage in intracellular Ca²⁺, but decreased O₂^? production capacity, mediated via the PKC pathway, in milk PMN. It is suggested that the defects in Ca²⁺ homeostasis in milk PMNs of lactating goats is partially attributable for the post-diapedesis functionality modifications.     

Effects of aerobic and anaerobic fluid collection on biochemical analysis of peritoneal fluid in healthy horses and horses with colic

Objective: To determine whether in healthy horses and those with colic, exposure of peritoneal fluid to room air affects values obtained on biochemical analysis. Study Design: Prospective study. Animals: Adult horses with a primary complaint of acute abdominal pain (n=29) and 12 healthy horses. Methods: Peritoneal fluid was aseptically collected under aerobic and anaerobic conditions. After collection, pH, PCO₂, PO₂, HCO₃^?, Na⁺, ionized Ca²⁺, K⁺, lactate, and glucose were immediately measured using a commercial blood gas analyzer. Biochemical variables were compared between aerobically and anaerobically obtained samples using a paired t‐test. Results: In healthy horses, peritoneal fluid samples collected under anaerobic conditions had higher PCO₂ and ionized Ca²⁺ and lower PO₂, HCO₃^?, and pH compared with samples exposed to air. No differences were observed for K⁺, Na⁺, glucose, and lactate. In horses with colic, samples collected anaerobically had higher PCO₂, ionized Ca²⁺, Na⁺, and glucose and lower PO₂, HCO₃^?, and pH value compared with samples exposed to air. No differences were observed for K⁺ and lactate. Conclusion: Exposure of peritoneal fluid to room air had a significant effect on pH, PCO₂, PO₂, and variables associated or dependent on changes in pH such as HCO₃^? and ionized Ca²⁺. Interpretation of biochemical analysis of peritoneal fluid may be influenced by sample collection method.     

Salt-Lick–Induced Soil Disturbance in the Teton Wilderness,USA

Manmade salt licks on public lands throughout the Rocky Mountain West have been created to attract large game for hunting purposes. This practice is both illegal and controversial, but is of particular importance in otherwise pristine wilderness landscapes. The impact of widespread saltlicks on public lands has never been quantified. This study was undertaken to examine the degree of change in soil physical and chemical properties caused by approximately 10–60 years of salt application in the Teton Wilderness of Wyoming, USA. A total of 27 sites were identified, surveyed, and paired with non–salt-affected control areas. Three replicate sampling points were located within each salt site and in each of the paired control areas. Soil samples from each site were analyzed for soil bulk density, soil salinity as electrical conductance (EC), pH, organic matter content, sodium absorption ratio (SAR), and exchangeable concentrations of sodium (Na⁺), potassium (K⁺), calcium (Ca²⁺), and magnesium (Mg²⁺). Salt-treated site centers were found to have elevated EC, bulk density, pH, SAR, and Na⁺ concentration compared to the no-salt controls. Salt-affected sites also contained decreased organic matter contents and decreased concentrations of Ca²⁺ and Mg²⁺. Observed differences were due to the addition of Na⁺ to the soil solum as well as direct effects of ungulates. Soil compaction appears to have a greater impact on plant establishment than the actual presence of NaCl. Salt licks established in wilderness areas habituate animals to localized zones causing extensive soil trampling and consumption of surface soils by grazing ungulates.     

Effects of dietary cation-anion balance on blood PH, acid base parameters, serum and urine mineral levels, and parathyroidhormone (PTH) in weanling horses.

This study demonstrated that the feeding of treatment diets with calculated dietary cation-anion balances (DCAB) of +370.43 (H) and -25.69 (L) did not have significant effects on blood pH, pCO₂, and HCO₃-. Serum Ca²⁺, P, Na⁺, and Cl^- as well as plasma PTH did not differ (P > .05) between the two treatment groups. Serum K⁺ was higher (P< .05) in horses fed diet H rather than diet L. The DCAB of the diet significantly affected urinary Ca²⁺, P, Na⁺, K⁺, and Cl^- excretion in the young growing horse. Urine Ca²⁺ and Cl- levels were higher (P < .01) in horses fed diet H versus diet L. Furthermore, levels of P, Na⁺, and K⁺ in the urine were higher (P < .01) in horses on diet H as opposed to diet L. Results of this study indicate that horses were able to maintain acid-base status regardless of diet. However, these data imply that growing horses consuming diets low in DCAB may be predisposed to abnormal bone mineralization due to the increase in calcium excretion which could lead to a weakening of the skeletal system.     

Glycine ameliorates mitochondrial dysfunction caused by ABT-199 in porcine oocytes

Mitochondria play an important role in controlling oocyte developmental competence. Our previous studies showed that glycine (Gly) can regulate mitochondrial function and improve oocyte maturation in vitro. However, the mechanisms by which Gly affects mitochondrial function during oocyte maturation in vitro have not been fully investigated. In this study, we induced a mitochondrial damage model in oocytes with the Bcl-2-specific antagonist ABT-199. We investigated whether Gly could reverse the mitochondrial dysfunction caused by ABT-199 exposure and whether it is related to calcium regulation. Our results showed that ABT-199 inhibited cumulus expansion, decreased the oocyte maturation rate and the intracellular glutathione (GSH) level, caused mitochondrial dysfunction, which was confirmed by decreased mitochondrial membrane potential (ΔΨm) and the expression of mitochondrial function-related genes PGC-1α, and increased reactiveoxygenspecies (ROS) levelsand the expression of apoptosis-associated genes Bax, Caspase-3, and Cyto C.More importantly, ABT-199-treated oocytes showed an increase in the intracellular free calcium concentration ([Ca²⁺]i) and had impaired cortical type 1 inositol 1,4,5-trisphosphate receptors (IP₃R1) distribution. Nevertheless, treatment with Gly significantly ameliorated mitochondrial dysfunction, oxidative stress, and apoptosis, and Gly also regulated [Ca²⁺]i levels and IP₃R1 cellular distribution, which further protects oocyte maturation in ABT-199-induced porcine oocytes.Taken together, our results indicate that Gly has a protective action against ABT-199-induced mitochondrial dysfunction in porcine oocytes.     

Accuracy of Potassium Supplementation of Fluids Administered Intravenously

Background

Potassium (K⁺) supplementation of isotonic crystalloid fluids in daily fluid therapy is commonly performed, yet its accuracy in veterinary medicine is undetermined.

Objective

To investigate the accuracy of K⁺ supplementation in isotonic crystalloid fluids.

Animals

None.

Methods

Observational study. 210 bags of fluid supplemented with KCl being administered to hospitalized dogs and cats intravenously (IV) were sampled over a 3‐month period. Measured K⁺ concentration ([K⁺]) was compared to the intended [K⁺] of the bag. In a second experiment, 60 stock fluid bags were supplemented to achieve a concentration of 20 mmol/L K⁺, mixed well and [K⁺] was measured. In another 12 bags of 0.9% NaCl, K⁺ was added without mixing the bag, and [K⁺] of the delivered fluid was measured at regular time points during constant rate infusion.

Results

The measured [K⁺] was significantly higher than intended [K⁺] (mean difference 9.0 mmol/L, range 6.5 to >280 mmol/L, P < .0001). In 28% of clinical samples measured [K⁺] was ≥5 mmol/L different than intended [K⁺]. With adequate mixing, K⁺ supplementation of fluids can be accurate with the mean difference between measured and intended [K⁺] of 0.7 (95% CI −0.32 to 1.7) mmol/L. When not mixed, K⁺ supplementation of 20 mmol/L can lead to very high [K⁺] of delivered fluid (up to 1410 mmol/L).

Conclusions and Clinical Importance

Inadequate mixing following K⁺ supplementation of fluid bags can lead to potentially life threatening IV infused [K⁺]. Standard protocols for K⁺ supplementation should be established to ensure adequate mixing.