Enhanced inactivation of avian influenza virus at ?20°C by disinfectants supplemented with calcium chloride or other antifreeze agents

Avian influenza outbreaks have occurred during winter months, and effective disinfection of poultry premises at freezing temperatures is needed. The commercial disinfectants Virkon and Accel, supplemented with an antifreeze agent [propylene glycol (PG), methanol (MeOH), or calcium chloride (CaCl₂)], were evaluated for their effectiveness in killing avian influenza virus (AIV) at −20°C or 21°C. An AIV suspension was applied to stainless steel disks, air-dried, and covered with a disinfectant or antifreeze agent for 5 to 30 min. Virkon (2%) and Accel (6.25%) with 30% PG, 20% MeOH, or 20% CaCl₂ inactivated 6 log₁₀ AIV within 5 min at −20°C and 21°C. At these temperatures PG and MeOH alone did not kill AIV, but the 20% CaCl₂ solution alone inactivated 5 log₁₀ AIV within 10 min. The results suggested that CaCl₂ is potentially useful to enhance the effectiveness of disinfection of poultry facilities after outbreaks of AIV infection in warm and cold seasons.     

Erythema multiforme associated with zonisamide in a dog

This report describes a dog that developed erythema multiforme in temporal association with administration of the sulphonamide‐based anticonvulsant drug zonisamide. Similar adverse drug reactions have been associated with sulphonamide antimicrobial drugs. Caution should be exercised when prescribing this medication for dogs with known hypersensitivity to sulphonamides.     

An evaluation of fresh gas flow rates for spontaneously breathing cats and small dogs on the Humphrey ADE semi‐closed breathing system

ObjectiveTo evaluate the fresh gas flow (FGF) rate requirements for the Humphrey ADE semi-closed breathing system in the Mapleson A mode; to determine the FGF at which rebreathing occurs, and compare the efficiency of this system to the Bain (Mapleson D) system in spontaneously breathing cats and small dogs.Study DesignProspective clinical study.AnimalsTwenty-five healthy (ASA score I or II) client-owned cats and dogs (mean ± SD age 4.7 ± 5.0 years, and body weight 5.64 ± 3.26 kg) undergoing elective surgery or minor procedures.MethodsAnaesthesia was maintained with isoflurane delivered via the Humphrey ADE system in the A mode using an oxygen FGF of 100 mL kg⁻¹ minute⁻¹. The FGF was then reduced incrementally by 5–10 mL kg⁻¹minute⁻¹ at approximately five-minute intervals, until rebreathing (inspired CO₂ >5 mmHg (0.7 kPa)) was observed, after which flow rates were increased. In six animals, once the minimum FGF at which rebreathing occurred was found, the breathing system was changed to the Bain, and the effects of this FGF delivery examined, before FGF was increased.ResultsRebreathing did not occur at the FGF recommended by the manufacturer for the ADE. The mean ± SD FGF that resulted in rebreathing was 60 ± 20 mL kg⁻¹minute⁻¹. The mean minimum FGF at which rebreathing did not occur with the ADE was 87 ± 39 mL kg⁻¹minute⁻¹. This FGF resulted in significant rebreathing (inspired CO₂ 8.8 ± 2.6 mmHg (1.2 ± 0.3 kPa)) on the Bain system.ConclusionsThe FGF rates recommended for the Humphrey ADE are adequate to prevent rebreathing in spontaneously breathing cats and dogs <15 kg.Clinical relevanceThe Humphrey ADE system used in the A mode is a more efficient alternative to the Bain system, for maintenance of gaseous anaesthesia in spontaneously breathing cats and small dogs.     

Impact of source tissue and ex vivo expansion on the characterization of goat mesenchymal stem cells

Background

There is considerable interest in using goats as models for genetically engineering dairy animals and also for using stem cells as therapeutics for bone and cartilage repair. Mesenchymal stem cells (MSCs) have been isolated and characterized from various species, but are poorly characterized in goats.

Results

Goat MSCs isolated from bone marrow (BM-MSCs) and adipose tissue (ASCs) have the ability to undergo osteogenic, adipogenic and chondrogenic differentiation. Cytochemical staining and gene expression analysis show that ASCs have a greater capacity for adipogenic differentiation compared to BM-MSCs and fibroblasts. Different methods of inducing adipogenesis also affect the extent and profile of adipogenic differentiation in MSCs. Goat fibroblasts were not capable of osteogenesis, hence distinguishing them from the MSCs. Goat MSCs and fibroblasts express CD90, CD105, CD73 but not CD45, and exhibit cytoplasmic localization of OCT4 protein. Goat MSCs can be stably transfected by Nucleofection, but, as evidenced by colony-forming efficiency (CFE), yield significantly different levels of progenitor cells that are robust enough to proliferate into colonies of integrants following G418 selection. BM-MSCs expanded over increasing passages in vitro maintained karyotypic stability up to 20 passages in culture, exhibited an increase in adipogenic differentiation and CFE, but showed altered morphology and amenability to genetic modification by selection.

Conclusions

Our findings provide characterization information on goat MSCs, and show that there can be significant differences between MSCs isolated from different tissues and from within the same tissue. Fibroblasts do not exhibit trilineage differentiation potential at the same capacity as MSCs, making it a more reliable method for distinguishing MSCs from fibroblasts, compared to cell surface marker expression.

Electronic supplementary material

The online version of this article (doi:10.1186/2049-1891-6-1) contains supplementary material, which is available to authorized users.     

Zinc supplementation strategies in feedlot heifers receiving an extended-release implant or an aggressive re-implant program

Two hundred eight Angus-crossbred heifers (291 ± 23 kg) from four sources were used in a randomized complete block design. The objective of the study was to determine the effects of implant strategy and Zn supplementation on performance, carcass characteristics, muscle fiber diameter, and mineral status of heifers. Heifers were assigned to a 2 × 2 factorial study for 168 d, and factors included Zn and implant (IMP). Heifers were supplemented Zn (mg/kg dry matter [DM]; ZnSO₄) at national (30; NRC) or industry (100; IND) recommendations. Implant strategies (Merck Animal Health, Madison, NJ) included extended-release Revalor-XH on day 0 (REV-XH; 20 mg estradiol + 200 mg trenbolone acetate) containing four uncoated pellets and six coated pellets or the uncoated implant Revalor-200 on day 0 and again on day 91 (REV-200; 20 mg estradiol + 200 mg trenbolone acetate). Heifers were blocked by weight within source to pens of five or six heifers per pen (nine pens per treatment). A corn silage-based diet was fed during the growing period (days 0–55) followed by transition to a corn-based finishing diet. Weights were taken consecutively on days −1/0, 55/56, and 167/168. Liver and muscle from the longissimus thoracis were collected from one heifer per pen on days −5, 14, 105, and 164. Data were analyzed via Mixed Procedure of SAS. Average daily gain (ADG) and liver mineral used Period as the repeated effect. Corresponding to periods of high hormone payout from each implant, days 0–28 and 91–120 ADG were greatest for REV-200, whereas REV-XH numerically peaked during days 56–91 (IMP × Period; P = 0.02). Day 91 IND body weight tended to be heavier (P = 0.06) and day 120 body weight was heavier (P = 0.05) than NRC heifers. No effect of Zn or IMP on final body weight was observed (P ≥ 0.21). Muscle fiber cross-sectional diameter on day 164 was greater (P = 0.05) in IND than NRC. Liver Mn concentrations decreased by day 14 regardless of implant, though days 105 and 164 concentrations were lesser for REV-200 than REV-XH (IMP × Period; P = 0.02). No effects of Zn, IMP, or the interaction were observed for carcass-adjusted gain to feed, days 0–168 DM intake, hot carcass weight, or ribeye area (P ≥ 0.11). The nominal differences in performance between implant strategies suggest that extended-release implants may be an effective implant strategy to replace re-implant programs in heifers, whereas the improved performance of heifers fed IND vs. NRC during times of peak hormone payout suggests a role for Zn in periods of rapid growth.     

Sulfate--a candidate for the missing essential factor that is required for the formation of protein haze in white wine

Protein haze formation in white wine is dependent on the presence of both wine protein and other unknown wine components, termed factor(s) X. The ability to reconstitute protein haze upon heating artificial model wine solutions (500 mg/L thaumatin, 12% ethanol, 4 g/L tartaric acid) to which candidate components were added was employed to identify factor(s) X. No protein haze was formed in the absence of additives. The individual or combined addition of caffeic acid, caftaric acid, epicatechin, epigallocatechin-O-gallate, gallic acid, or ferulic acid at typical white wine concentrations did not generate protein haze. However, PVPP fining of commercial wines resulted in a reduction in protein haze, suggesting that phenolic compounds may play a modulating role in haze formation. To elucidate the nature of the unknown factor(s) wine was fractionated and fractions were back-added to model wine and tested for their essentiality. Wine fractions were generated by ultrafiltration, reverse-phase chromatography, and mixed-mode anion-exchange and reverse-phase chromatography. The only purified fraction containing the essential component(s) was free of phenolic compounds, and analysis by mass spectrometry identified sulfate anion as the dominant component. Reconstitution with KHSO4 using either commercially available thaumatin or wine proteins confirmed the role of sulfate in wine protein haze formation. The two main wine proteins, thaumatin-like protein and chitinase, differed in their haze response in model wines containing sulfate. Other common wine anions, acetate, chloride, citrate, phosphate, and tartrate, and wine cations, Fe(2+/3+) and Cu(+/2+), when added at typical white wine concentrations were not found to be essential for protein haze formation.