A meta-analysis of fumarate effects on methane production in ruminal batch cultures

The objective of this study was to understand the effects of fumarate addition on methane (CH4) and VFA production in the rumen through a meta-analysis of its effects on ruminal batch cultures. Because the reduction of fumarate to succinate can draw electrons away from ruminal methanogenesis, fumarate has been studied as a potential feed additive to decrease CH4 production in ruminants. The average decrease in CH4 in batch cultures was of 0.037 micromol/micromol of added fumarate, which is considerably lower than 0.25 micromol/micromol, the decrease predicted from the stoichiometry of the reactions involved. One reason that fumarate was not effective at decreasing CH4 in batch cultures was that only an average of 48% of added fumarate appeared to be converted to propionate. Secondly, the incorporation of reducing equivalents in the conversion of fumarate to propionate was almost entirely offset by their release from an average of 20% of added fumarate that appeared to be converted to acetate. Thermodynamic calculations indicated that the conversion of added fumarate to both propionate and acetate was feasible. Fumarate appears to be more effective in decreasing CH4 production and increasing propionate in continuous culture than in batch culture. This suggests that microbial adaptation to fumarate metabolism can be important. Variation in populations of fumarate-reducers, methanogens, and protozoa could all be involved. Fumarate supplementation for an extended period may result in the amplification of otherwise small populations of fumarate-reducers. Addition of some of these organisms may be helpful to improve fumarate conversion to propionate. Strategies based on enhancing the rumen's capacity to convert fumarate to propionate by maintaining a low fumarate concentration have been effective. Thermodynamic considerations should be taken into account when designing strategies for CH4 abatement through the addition of external electron acceptors.     

Cloning and sequence analysis of the complementary DNA for feline preproparathyroid hormone

OBJECTIVES: To clone and sequence the cDNA for feline preproparathyroid hormone (preproPTH) and to compare that sequence with other known parathyroid hormone (PTH) sequences. SAMPLE POPULATION: Parathyroid glands from 1 healthy cat. PROCEDURES: A cDNA library was constructed in lambda phage from feline parathyroid gland mRNA and screened with a radiolabeled canine PTH probe. Positive clones were sequenced, and nucleic acid and deduced amino acid sequences were analyzed and compared with known preproPTH and PTH sequences. RESULTS: Screening of approximately 2 X 10(5) recombinant plaques revealed 3 that hybridized with the canine PTH probe; 2 clones comprised the complete sequence for feline preproPTH. Feline preproPTH cDNA consisted of a 63-base pair (bp) 5'-untranslated region (UTR), a 348-bp coding region, and a 326-bp 3'-UTR. The coding region encoded a 115-amino acid peptide. Mature feline PTH consisted of 84 amino acids. Amino acid sequence analysis revealed that feline PTH was > 83% identical to canine, bovine, swine, equine, human, and macaque PTH and 69, 71, and 44% identical to mouse, rat, and chicken PTH, respectively. Within the region responsible for hormonal activity (amino acids 1 to 34), feline PTH was > 79% identical to other mammalian PTH sequences and 64% identical to the chicken sequence. CONCLUSIONS AND CLINICAL RELEVANCE: The amino acid sequence of PTH is conserved among mammalian species. Knowledge of the cDNA sequence for feline PTH may be useful to investigate disturbances of calcium metabolism and alterations in PTH expression in cats.     

In vitro dose-dependent effects of enrofloxacin on equine articular cartilage.

OBJECTIVE: To determine whether enrofloxacin has detrimental, dose-dependent effects on equine articular cartilage in vitro. ANIMALS: Cartilage explants were developed from 6 healthy horses between 0 and 96 months old. PROCEDURE: Patellar cartilage explants were incubated in 5 concentrations of enrofloxacin (2 microg/ml, 10 microg/ml, 1,000 microg/ml, 10,000 microg/ml, and 50,000 microg/ml) for 72 hours. Proteoglycan synthesis (Na35SO4 incorporation for 24 hours), proteoglycan degradation (Na35SO4 release for 72 hours), endogenous proteoglycan content (dimethylmethlene blue assay), and total protein content were determined. Cartilage explants were evaluated by use of histomorphologic and histomorphometric techniques (toluidine blue stain) for cytologic and matrix characteristics. Quantitative data were analyzed with a one-way ANOVA to compare results among various enrofloxacin concentration groups and the control group. A general linear model was used to determine whether age had an effect. RESULT: Proteoglycan synthesis was excellent in control specimens and in specimens incubated in low concentrations of enrofloxacin (2 microg/ml and 10 microg/ml). High concentrations of enrofloxacin (> 1,000 microg/ml) effectively eliminated proteoglycan synthesis regardless of horse age. Proteoglycan degradation at low concentrations (2 microg/ml and 10 microg/ml) was not different than control. High concentrations of enrofloxacin (> 1,000 microg/ml) caused significant degradation. Different concentrations of enrofloxacin did not affect endogenous proteoglycan. High concentrations of enrofloxacin were associated with a significant increase in number of pyknotic nuclei. CONCLUSION: Concentrations of enrofloxacin that might be achieved following systemic administration did not suppress chondrocyte metabolism in vitro. High concentrations of enrofloxacin (> 1,000 microg/ml) were toxic to chondrocytes.     

Paranaella luquei gen. et sp. n. (Monogenea: Microcotylidae), a new parasite of Brazilian catfishes

Paranaella, a new microcotyline monotypic genus, is erected to accommodate Paranaella liquei sp. n., parasite of gill filaments from Hypostomus sp., Hypostomus regani (Ihering) and Rhinelepis aspera Spix et Agassiz (Loricariidae) from the Parani River, Brazil. The new genus is most closely related to Microcotyle Van Beneden et Hesse, 1863, Diplostamenides Unnithan. 1971 and Solostamenides Unnithan, 1971. From Microcotyle it differs mainly by having the genital atrium formed by a muscular ring with a concentric row of numerous elongate and straight spines; from Diplostamenides it can be distinguished by the unarmed and not differentiated cirrus and from Solostamenides it differs by the single vaginal pore and absence of larval hooks.     

Evaluation of a point-of-care test for canine C-reactive protein

Background: In veterinary medicine, there is increasing interest in measuring C‐reactive protein (CRP) as a tool for diagnosis and monitoring of inflammatory diseases. Reported CRP concentrations for healthy dogs have ranged from 0 to 8.9 mg/L. Objectives: The aims of this study were to evaluate a canine‐specific point‐of‐care (POC) lateral flow immunoassay for qualitative CRP measurement in healthy and diseased dogs and to compare results with those obtained by a quantitative ELISA. Methods: Blood samples from 73 client‐owned dogs were available for testing: 16 healthy dogs and 57 dogs with a variety of infectious, inflammatory, or neoplastic diseases. CRP was measured in heparinized whole blood samples and serum with the TECOmedical Dog CRP‐visual POC test. A red line develops in the POC device if CRP is ≥5 mg/L, and results are scored as negative or positive. An ELISA validated previously for canine serum was used as the reference method. Results: For all dogs, serum CRP concentrations measured by the ELISA ranged from 0.1 to ≥350 mg/L (median=38 mg/L). Percentages of the CRP POC test results that agreed with the ELISA results were 98.6% for whole blood and 97.3% for serum samples. For serum samples, sensitivity of the POC test was 96.4% and specificity was 81.3%. For whole blood, sensitivity was 94.7% and specificity was 93.8%. Conclusions: The POC test had very good agreement with the ELISA test and had high sensitivity and specificity; therefore, it can be used as a qualitative test to screen for increases in CRP concentrations.     

Molecular epidemiology of environmental MRSA at an equine teaching hospital: introduction,circulation and maintenance

The role that environmental contamination might play as a reservoir and a possible source of Methicillin-resistant Staphylococcus aureus (MRSA) for patients and personnel at equine veterinary hospitals remains undefined, as the environment has only been monitored during outbreaks or for short periods. Therefore, the objectives of this study were to determine the monthly presence, distribution, and characteristics of environmental MRSA at an equine hospital, and to establish patterns of contamination over time using molecular epidemiological analyses. For this purpose, a yearlong active MRSA surveillance was performed targeting the environment and incoming patients. Antimicrobial susceptibility testing, SCCmec typing, PFGE typing, and dendrographic analysis were used to characterize and analyze these isolates. Overall, 8.6% of the surfaces and 5.8% of the horses sampled were positive for MRSA. The most common contaminated surfaces were: computers, feed-water buckets, and surgery tables-mats. Ninety percent of the isolates carried SCCmec type IV, and 62.0% were classified as USA500. Molecular analysis showed that new pulsotypes were constantly introduced into the hospital throughout the year. However, maintenance of strains in the environment was also observed when unique clones were detected for 2 consecutive months on the same surfaces. Additionally, pulsotypes were circulating throughout several areas and different contact surfaces of the hospital. Based on these results, it is evident that MRSA is constantly introduced and frequently found in the equine hospital environment, and that some contact surfaces could act as “hot-spots”. These contaminated surfaces should be actively targeted for strict cleaning and disinfection as well as regular monitoring.     

Alterations in serum parathyroid hormone and electrolyte concentrations and urinary excretion of electrolytes in horses with induced endotoxemia

Hypocalcemia and hypomagnesemia are common in horses with sepsis and endotoxemia. We hypothesize that endotoxemia triggers a systemic inflammatory response that results in hypocalcemia and hypomagnesemia. The goal of this study was to determine the effect of endotoxin (lipopolysaccharide [LPS]) administration to healthy horses on serum parathyroid hormone (PTH), ionized calcium (Ca2+) and total calcium (tCa), ionized magnesium (Mg2+) and total magnesium (tMg), phosphate (Pi), potassium (K+), sodium (Na+), chloride (Cl-), and insulin concentrations, and on the urinary excretion of these electrolytes. Twelve mares were infused with Escherichia coli LPS (30 ng/kg/h i.v.) for 1 hour. Six mares were infused with saline (controls). In LPS-infused horses, heart rate increased significantly from (mean +/- SD) 40.0 +/- 1.3 to 70.0 +/- 9.0 beats/min, respiratory rate from 12.7 +/- 1.0 to 21.1 +/- 3.0 breaths/min, body temperature from 37.4 +/- 0.3 to 38.9 +/- 0.6 degrees C, and tumor necrosis factor-alpha concentrations from 6.6 +/- 3.5 to 507 +/- 260 pg/mL (P < .05). White blood cell count decreased significantly from 7570 +/- 600 to 1960 +/- 560 cells/ microL. Serum concentrations of Ca2+ decreased from 6.5 +/- 0.3 to 6.0 +/- 0.3 mg/dL, of Mg2+ from 0.53 +/- 0.06 to 0.43 +/- 0.04 mM, of tMg from 0.78 +/- 0.05 to 0.62 +/- 0.08 mM, of K+ from 4.3 +/- 0.4 to 3.0 +/- 0.5 mEq/L, and of Pi from 3.4 +/- 0.5 to 1.7 +/- 0.5 mg/dL (all P < .05). PTH increased significantly from 1.3 +/- 0.4 to 6.0 +/- 5.2 pM; however, in some horses (n=2), PTH did not increase despite hypocalcemia. Insulin increased significantly from 9.4 +/- 3.6 to 50.5 +/- 9.6 microIU/mL (n=3). Urinary fractional excretion of Ca2+ decreased significantly from 4.7 +/- 1.4 to 1.7 +/- 1.2%, of Mg2+ from 36.6 +/- 6.5 to 11.7 +/- 7.3%, and of K+ from 37.9 +/- 11.3 to 17.7 +/- 6.2%. Fractional excretion of Pi increased from 0.02 +/- 0.02 to 0.14 +/- 0.07% and of Na+ from 0.26 +/- 0.13% to 1.2 +/- 0.5%. No changes were found in serum tCa, Na+, and Cl- concentrations. In conclusion, endotoxemia in horses resulted in electrolyte abnormalities that included hypocalcemia, hypomagnesemia, hypokalemia, hypophosphatemia, and increased serum PTH and insulin concentrations.