Proton pumps in the fish gill and kidney

The proton pump or vacuolar type H⁺-ATPase is an oligomeric protein responsible for electrogenic H⁺ secretion in a variety of acid-secreting epithelia. Recently, the proton pump was identified in both the gill and kidney of freshwater-adapted rainbow trout (Oncorhynchus mykiss). Using immunocytochemistry, H⁺-ATPase has been localized in the pavement cells and chloride cells of the lamellar epithelium. During periods of internal acidosis, there is a marked increase in the expression of the branchial proton pump as identified by Western analysis, immunocytochemistry and in situ hybridization. This augmented expression of proton pumps occurs concomitantly with a marked increase in branchial acid excretion and Na⁺ uptake. Immunocytochemical studies suggest that the pavement cell, rather than the chloride cell, is the predominant site of acid excretion during periods of acidosis. These findings are consistent with the notion that in freshwater teleosts, Na⁺ uptake and H⁺ excretion are linked via the coupling of the electrogenic proton pump to apical membrane Na⁺ channels. This mechanism may be controlled by hormones including cortisol and/or growth hormone. The fish kidney plays an important role in regulating acidosis via the re-absorption of filtered HCO₃ ^-. Recently, we have demonstrated using Western analysis and immunocytochemistry, the presence of proton pump in rainbow trout kidney and observed increased H⁺-ATPase expression during respiratory acidosis. These new findings suggest a role for the renal proton pump in acid-base regulation.     

Histological changes in insulin-immunoreactive pancreatic β-cells,and suppression of insulin secretion and somatotrope activity in brook trout (Salvelinus fontinalis) maintained on reduced food intake or exposed to acidic environment

Immature brook trout (Salvelinus fontinalis) were randomly divided into a pH control, a pH and food control and an acid-stressed group. Fish in the first two groups were held at neutral pH and those in the last group were maintained at pH 4.2 for up to two months. The food supply to the pH and food control group was restricted to simulate the reduction in food intake demonstrated for acid-stressed trout. Plasma insulin levels were significantly decreased from 5–20 ng/ml to 1–2 ng/ml and plasma cortisol levels were significantly increased from 5–10 ng/ml to as high as 70 ng/ml in the acid-stressed brook trout. Concomitantly, a significant decrease of 21–39% in the proportion (volume density) of insulin immunoreactive -cells was observed within the principal pancreatic islets. Somatic growth was stunted and ultrastructural morphometry revealed the suppression of somatotrope secretory activity in the acid-stressed fish. Restriction of food supply induced a smaller but still significant decrease in circulating levels of insulin which was however not accompanied by a reduction in insulin immunoreactive -cells. The rise in plasma cortisol levels was not significant, and the plasma levels of glucose and protein were unaffected. Nevertheless, somatotrope secretory activity was suppressed and somatic growth was stunted. This study demonstrates for the first time the complexity of the endocrine response to acid stress and that some of the response to acid stress can be attributed to the lowering of food intake.     

A cross-sectional pilot study to estimate the prevalence of and risk factors for leptospirosis in South-Western Victorian dairy herds, 2017

Leptospirosis is a zoonosis, found worldwide, affecting many species of animals. We conducted a cross-sectional study to estimate the prevalence of Leptospira borgpetersenii sv Hardjo and Leptospira interrogans sv Pomona in cattle in dairy herds in South-Western Victoria, Australia. Fifty-three herds were enrolled in the study. Urine samples were collected from 15 late-lactation cows in each herd. A questionnaire was provided to herd managers at the time of each herd visit, asking them to describe the methods they used for controlling leptospirosis, including vaccination. Urine samples were pooled at the herd level and tested for leptospira spp. using real time PCR. Urine samples from individual cows within the positive pooled samples were then tested for Leptospira Hardjo and Leptospira Pomona using qPCR. Four of the 53 herds showed positive leptospirosis results giving an apparent prevalence of 8 (95% CI 2–18) leptospira-positive herds per 100 herds at risk. Based on the 53 completed questionnaires, leptospirosis vaccination programs were not compliant with label directions in 36 of the 52 vaccinated herds: 69 (95% CI 55–81) of 100 herd managers that routinely vaccinated for leptospirosis did not comply with label directions. One herd was completely unvaccinated. Based on our findings, we estimate that approximately 10% of dairy farms in South-Western Victoria are likely to be infected with leptospirosis. While most herds are vaccinating for leptospirosis, most are not doing so according to label directions. We conclude that herd managers need to be better educated regarding leptospirosis vaccination programs.     

Absorption by sheep of dieldrin from contaminated soil

Objective To study the accumulation of dieldrin residues in sheep from ingestion of contaminated soils was studied in two experiments.
Design A controlled feeding study of sheep fed contaminated soils of different type at varying intervals.
Animals and procedure Thirty-four 2-years-old wethers were divided into four groups (one control sheep only) and fed water-soluble dieldrin or soil contaminated with aldrin and dieldrin at varying intervals in the first study. In a second study 34 similar sheep were divided into four treatments with one being a control. Sheep were fed sandy, high clay or high organic matter soils with similar dieldrin and aldrin concentrations.
Results In the first study the concentration of dieldrin in the body fat of sheep dosed with dieldrin-contaminated soil was about half that in the body fat of sheep dosed with an equivalent amount of water-soluble dieldrin. The concentration of dieldrin was almost the same in sheep fed 500 μg of total dieldrin per day as it was in sheep fed 5000 μg every tenth day, over a 50-day period. In the second experiment sheep accumu-lated nearly three times as much pesticide from a soil with a high organic matter content, and about four times as much from a soil with a high clay content, as from a sandy soil with the same dieldrin content, over a 100-day period. The half-life of dieldrin in the fat of all sheep varied between 96 and 116 days after sheep ceased ingesting contaminated soil.
Conclusions Dieldrin concentrations in the fat of sheep that consume dieldrin contaminated soil fall within 10 days of removal from the source of contamination. However, dieidrin accumulates in the wool of sheep that consume dieldrin-contaminated soil.     

Short-term anaesthesia in piglets using a liquid injection, rebreathing, inhaler device

An economical anaesthetic technique of short duration that can be administered to piglets in the field is desirable for humanitarian reasons, for castration, tail docking or other brief procedures. Using the principles of anaesthetic uptake and distribution, an inhaler was developed to vaporize and administer isoflurane to piglets. The inhaler design consisted of a mask, vaporization chamber and a rebreathing bag. A stopcock provided access for injection of liquid isoflurane onto a wick contained in the vaporization chamber. Inspiratory and expiratory flow of air over the wick enhanced anaesthetic vaporization. The amount of liquid isoflurane required for induction and 2–3 minutes of surgical anaesthesia was calculated using the square root of time model proposed by Lowe & Ernst (1981) for liquid injection, closed circuit anaesthesia in people. Calculations were based on an assumed MAC of 1.4% and the achievement of a target alveolar concentration of 1.3 MAC to provide a surgical plane of anaesthesia. The appropriate isoflurane concentrations in the mask, inhaler, rebreathing bag and the piglet's FRC and tissues were calculated. Original calculations were based on metabolic size (BW^0.75) and then converted to weight (kg). Based on the piglet's scale weight, the total microliters of liquid isoflurane required were formulated into a table for field use. Isoflurane was injected into the inhaler stopcock followed by oxygen to fill the rebreathing bag and initiate vaporization. After the mask was placed over the piglet's nose a slide switch was activated to allow gases to move in and out of the inhaler and rebreathing bag. Fifty‐seven male piglets weighing (mean ± SD 3.0 ± 0.7 kg) and aged 7.7 ± 1.0 days were randomly selected to receive anaesthesia prior to castration. Remaining littermates served as controls for assessing morbidity or mortality. The time to induction, recovery and total anaesthetic time were measured. The Pe ′CO₂ was measured at the piglet's nostril immediately after the mask was removed at the end of the surgical procedure. Data were analysed in SAS using the Proc Mixed procedure. Inductions were rapid, 47 ± 9 seconds, generally with minimal or no resistance. The duration of surgery was 1–2 minutes. Anaesthesia was adequate and recovery was rapid, 122 ± 44 seconds. Total time from start to standing was 260 ± 51 seconds. The Pe ′CO₂ was 5.2 ± 1.1 kPa (39.4 ± 8.4 mm Hg). There was no morbidity or mortality associated with either group of piglets. After piglets were standing and mobile, they were returned to the sow and other littermates, where they immediately started nursing and were indistinguishable from littermates except by determination of ear notch number. This technique provides safe, rapid anaesthesia and recovery that is appropriate for use by veterinarians for brief field procedures.     

Is a purpose of REM sleep atonia to help regenerate intervertebral disc volumetric loss?

The nature of atonia in sleep continues to be enigmatic. This article discusses a new hypothesis for complete core muscle relaxation in REM sleep, suggesting a bottom-up recuperative perspective. That is, does the atonia in REM sleep provide a utility to help restore the mechanobiology and respective diurnal intervertebral disc hydraulic loss? By combining the effects of gravity with current compressive concepts in spinal stability, this article looks at vertebral approximation as a deleterious experience with an intrinsic biological need to keep vertebrae separated. Methods using polysomnography and recumbent MRI are discussed.     

The lack of effect of isoflavones on high-density lipoprotein cholesterol concentrations in adolescent boys: a 6-week randomised trial

BACKGROUND: A substantial fall in high-density lipoprotein cholesterol (HDL-C) during puberty in boys, but not girls, has been reported in Western populations. The fall in boys is believed to be due to hormonal changes - androgens have been shown to be associated with lower HDL-C, whereas oestrogens are associated with higher HDL-C. The fall in HDL-C during puberty was not observed, however, in a study of Moslem boys in Israel, nor in a group of Japanese boys. A diet high in phyto-oestrogens may account for the lack of a fall in HDL-C in these populations. OBJECTIVE: To examine the effect of dietary supplementation with phyto-oestrogens on the HDL-C concentration of adolescent boys from a Western population. We hypothesised that dietary supplementation of 50 mg of the isoflavones daidzein and genistein would produce a 12% higher HDL-C concentration than in controls at the end of a 6-week intervention period. DESIGN: A randomised controlled trial. SETTING: Hellyer College in Burnie (Tasmania, Australia). SUBJECTS: Adolescent boys (aged 16-18 years) were recruited through a letter sent to parents. A total of 132 eligible participants enrolled and five subjects withdrew from the trial. RESULTS: No significant increase in HDL-C was observed in the treatment group (-0.02 mmol l-1, standard error (SE)=0.03, P = 0.53) or the placebo group (0.05 mmol l-1, SE = 0.03, P = 0.11). CONCLUSIONS: Factors other than isolated dietary isoflavones may be responsible for the lack of fall in HDL-C during puberty in Japanese and Moslem boys.     

Addition of Recombinant Human Growth Hormone to In Vitro Maturation Medium of Bovine Oocytes

The aim of this study was to increase the bovine embryonic development rate, adding recombinant human growth hormone (rhGH) to maturation medium of bovine oocytes. Oocytes were matured for 24 h in TCM 199 Earle's salts and five treatments were developed: T1, 0.01 IU/ml of recombinant human follicle stimulating hormone (rhFSH); T2, 0.01 IU/ml of rhFSH + 100 ng/ml of rhGH; T3, 0.01 IU/ml of rhFSH + 1000 ng/ml of rhGH; T4, 100 ng/ml of rhGH; and T5, 1000 ng/ml of rhGH at 39 degrees C and 5% of CO(2) in air and saturated humidity. In vitro fertilization from cumulus-oocyte complexes was conducted in TALP-Fert medium (18-22 h) and spermatozoa were selected by Percoll gradient. Zygotes were incubated in SOFaaci medium in 5% of CO(2) in air, 5% of O(2) at 39 degrees C and saturated humidity for 11 days. There was no statistical difference in cleavage rate and embryo production on day 7 and day 9 among treatments. However, the hatching rate increased significantly in the T4 and T5 treatments (11.0 and 12.8%, respectively), compared with the T1 treatment (4.6%) (p < 0.05). Therefore, the rhGH addition to the oocyte maturation medium showed beneficial effects on the hatching rate of in vitro-produced bovine embryos.