Cell growth characteristics and differentiation frequency of adherent equine bone marrow-derived mesenchymal stromal cells: adipogenic and osteogenic capacity

OBJECTIVES: To characterize equine bone marrow (BM)-derived mesenchymal stem cell (MSC) growth characteristics and frequency as well as their adipogenic and osteogenic differentiation potential. STUDY DESIGN: In vitro experimental study. ANIMALS: Foals (n=3, age range, 17-51 days) and young horses (n=5, age range, 9 months to 5 years). METHODS: Equine MSCs were harvested and isolated from sternal BM aspirates and grown up to passage 10 to determine cell-doubling (CD) characteristics. Limit dilution assays were performed on primary and passaged MSCs to determine the frequency of colony-forming units with a fibroblastic phenotype (CFU-F), and the frequency of MSC differentiation into adipocytes (CFU-Ad) and osteoblasts (CFU-Ob). RESULTS: Initial MSC isolates had a lag phase with a significantly longer CD time (DT=4.9+/-1.6 days) compared with the average DT (1.4+/-0.22 days) of subsequent MSC passages. Approximately 1 in 4224+/-3265 of the total nucleated BM cells displayed fibroblast colony-forming activity. Primary MSCs differentiated in response to adipogenic and osteogenic inductive conditions and maintained their differentiation potential during subsequent passages. CONCLUSIONS: The frequency, in vitro growth rate, and adipogenic and osteogenic differentiation potential of foals and young adult horses are similar to those documented for BM MSCs of other mammalian species. CLINICAL RELEVANCE: The results have direct relevance to the use of BM as a potential source of adult stem cells for tissue engineering applications in equine veterinary medicine.     

Formulation with CpG ODN enhances antibody responses to an equine influenza virus vaccine

Previous studies have shown that protection against equine influenza virus (EIV) is partially mediated by virus-specific IgGa and IgGb. In this study we tested whether addition of a CpG ODN formulation to a commercial killed virus vaccine would enhance EIV-specific IgGa and IgGb antibody responses, and improve protection against an experimental EIV challenge. Thirty na?ve horses were assigned to one of three groups and vaccinated as follows: 10 were given vaccine (Encevac TC4, Intervet Inc.) alone, 10 were given vaccine plus 0.25 mg CpG ODN 2007 formulated with 30% Emulsigen (CpG/Em), and 10 controls were given saline. All horses were challenged with live virus 12 weeks after the final vaccination. Antibody responses were tested by single radial hemolysis (SRH) and ELISA, and protection was evaluated by determination of temperature, coughing, and clinical scores. Killed virus vaccine combined with CpG/Em induced significantly greater serologic responses than did the vaccine alone. All antibody isotypes tested increased after the addition of CpG/Em, although no shift in relative antibody isotypes concentrations was detected. Vaccination significantly improved protection against challenge but the differences between the two vaccine groups were not statistically significant. This study is the first demonstration that CpG/Em enhances antigen-specific antibody responses in horses and supports its potential to be used as an adjuvant for vaccines against equine infections.     

Long-term efficacy of trilostane administered twice daily in dogs with pituitary-dependent hyperadrenocorticism

Trilostane is considered an efficacious and safe medication for canine pituitary-dependent hyperadrenocorticism (PDH). Its recommended frequency of administration is once daily. In this prospective study, the efficacy, toxicity, and long-term outcome of trilostane administered twice daily per os were evaluated in 44 dogs with PDH. Mean initial dose was 3.1 mg/kg q 12 hours, and mean final dose was 3.2 mg/kg q 12 hours. The final total daily dose was lower than previously reported for once-daily administration. The mean survival time for affected dogs was 930 days.     

Effect of changes in ionized calcium concentration in arterial blood and metabolic acidosis on the arterial partial pressure of oxygen in dogs

OBJECTIVE: To evaluate the effects of metabolic acidosis and changes in ionized calcium (Ca2+) concentration on PaO2 in dogs. ANIMALS: 33 anesthetized dogs receiving assisted ventilation. PROCEDURE: Normal acid-base status was maintained in 8 dogs (group I), and metabolic acidosis was induced in 25 dogs. For 60 minutes, normocalcemia was maintained in group I and 10 other dogs (group II), and 10 dogs were allowed to become hypercalcemic (group III); hypocalcemia was then induced in groups I and II. Groups II and IV (5 dogs) were treated identically except that, at 90 minutes, the latter underwent parathyroidectomy. At intervals, variables including PaO2, Ca2+ concentration, arterial blood pH (pHa), and systolic blood pressure were assessed. RESULTS: In group II, PaO2 increased from baseline value (96 +/- 2 mm Hg) within 10 minutes (pHa, 7.33 +/- 0.001); at 60 minutes (pHa, 7.21 +/- 0.02), PaO2 was 108 +/- 2 mm Hg. For the same pHa decrease, the PaO2 increase was less in group III. In group I, hypocalcemia caused PaO2 to progressively increase (from 95 +/- 2 mm Hg to 104 +/- 3 mm Hg), which correlated (r = -0.66) significantly with a decrease in systolic blood pressure (from 156 +/- 9 mm Hg to 118 +/- 10 mm Hg). Parathyroidectomy did not alter PaO2 values. CONCLUSIONS AND CLINICAL RELEVANCE: Induction of hypocalcemia and metabolic acidosis each increased PaO2 in anesthetized dogs, whereas acidosis-induced hypercalcemia attenuated that increase. In anesthetized dogs, development of metabolic acidosis or hypocalcemia is likely to affect ventilatory control.     

Reversible reduction of oxygen to peroxide facilitated by molecular recognition

Generation of soluble sources of peroxide dianion (O(2)(2-)) is a challenge in dioxygen chemistry. The oxidizing nature of this anion renders its stabilization in organic media difficult. This Report describes the chemically reversible reduction of oxygen (O(2)) to cryptand-encapsulated O(2)(2-). The dianion is stabilized by strong hydrogen bonds to N-H groups from the hexacarboxamide cryptand. Analogous stabilization of peroxide by hydrogen bonding has been invoked recently in crystalline saccharide and protein systems. The present peroxide adducts are stable at room temperature in dimethyl sulfoxide (DMSO) and N,N'-dimethylformamide (DMF). These adducts can be obtained in gram quantities from the cryptand-driven disproportionation reaction of potassium superoxide (KO(2)) at room temperature.     

The impact of feed withdrawal on quality, safety, yield of pocessed chickens

一旦肉鸡达到上市体重并准备送入屠宰厂之时,肉品的加工处理就已经开始了.在这个时候,为了出空肉鸡肠道中的排泄物以防止在切除内脏时所发生的污染而必须采取禁食措施.否则,会影响肉鸡胴体预冷前的质量安全.     

Hunger and health among undocumented Mexican migrants in a US urban area

OBJECTIVES: To measure the occurrence and correlates of hunger and to evaluate the association between hunger and three health indicators among undocumented Mexican immigrants. DESIGN: Non-probability cross-sectional sample. SETTING: Neighbourhoods within New York City. SUBJECTS: Four hundred and thirty-one undocumented Mexican immigrants living in the USA. RESULTS: Hunger was indicated by approximately 28% of respondents. In a multivariate model, working as a day labourer was associated with hunger (odds ratio (OR) 3.33, 95% confidence interval (CI) 1.83-6.06) while receiving public assistance protected against hunger (OR 0.23, 95% CI 0.06-0.88). In multivariate models, respondents who reported experiencing hunger also reported poorer overall health (OR 1.69, 95% CI 0.95-3.02) and more days of poor mental (P = 0.045) and physical health (P < 0.0001). Greater amount of time lived in the USA was also associated with worse overall health (P = 0.054) and more days of poor mental and physical health (P < 0.01). CONCLUSIONS: The present study shows that food insecurity and hunger may be problems among undocumented migrants living in the USA. Uncertain and unpredictable work schedules and limited access to public assistance may contribute to high levels of hunger, which in turn may also negatively affect mental and physical health. Increasing amount of time lived in the USA is also associated with poorer health indicators. Programmes that provide undocumented migrants with emergency access to resources may reduce food insecurity and lead to improved health outcomes among this vulnerable population.     

Genetic parameters for litter traits at different parities in purebred Landrace and Yorkshire pigs

Comparison of the multi‐trait animal model and the traditional repeatability model was carried out using data obtained from 6,424 Landrace and 20,835 Yorkshire sows farrowed from January 2000 to April 2018 in order to estimate genetic parameters for litter traits at different parities. Specifically, records of the total number born (TNB), number born alive (NBA), total number of mortality (MORT), number of stillborn (NSB) and number of mummified pigs (MUM) were used. Although results showed the heterogeneity of heritability for litter traits at different parities, the mean heritability estimates from the multi‐trait model were found to be higher than those of the repeatability model for all traits in both pig breeds. In terms of genetic correlation between parities, a slight difference in genetic control in the first parity was noted for TNB and NBA in Landrace and Yorkshire pigs. The correlation between the first parity and later parities ranged from 0.48 to 0.74 for TNB and NBA in both breeds. Moreover, genetic correlation between parities for MORT and NSB was observed to be high for parities higher than 2 in Yorkshire pigs. For MUM, genetic correlation between the first and other parities was generally low in both breeds, indicating that culling pigs on the basis of MUM at the first parity could probably be unreasonable. Overall, the results of this study suggest that the multi‐trait approach for litter size traits is useful for the accurate estimation of genetic parameters.     

Genetic diversity and phylogenetic relationships between Leishmania infantum from dogs,humans and wildlife in south‐east Spain

Leishmania infantum causes human and canine leishmaniosis. The parasite, transmitted by phlebotomine sand flies, infects species other than dogs and people, including wildlife, although their role as reservoirs of infection remains unknown for most species. Molecular typing of parasites to investigate genetic variability and evolutionary proximity can help understand transmission cycles and designing control strategies. We investigated Leishmania DNA variability in kinetoplast (kDNA) and internal transcribed spacer 2 (ITS2) sequences in asymptomatically infected wildlife (n = 58) and symptomatically and asymptomatically infected humans (n = 38) and dogs (n = 15) from south‐east Spain, using single nucleotide polymorphisms (SNPs) and in silico restriction fragment length polymorphism (RFLP) analyses. All ITS2 sequences (n = 76) displayed a 99%–100% nucleotide identity with a L. infantum reference sequence, except one with a 98% identity to a reference Leishmania panamensis sequence, from an Ecuadorian patient. No heterogeneity was recorded in the 73 L. infantum ITS2 sequences except for one SNP in a human parasite sequence. In contrast, kDNA analysis of 44 L. infantum sequences revealed 11 SNP genotypes (nucleotide variability up to 4.3%) and four RFLP genotypes including B, F and newly described S and T genotypes. Genotype frequency was significantly greater in symptomatic compared to asymptomatic individuals. Both methods similarly grouped parasites as predominantly or exclusively found in humans, in dogs, in wildlife or in all three of them. Accordingly, the phylogenetic analysis of kDNA sequences revealed three main clusters, two as a paraphyletic human parasites clade and a third including dogs, people and wildlife parasites. Results suggest that Leishmania infantum genetics is complex even in small geographical areas and that, probably, several independent transmission cycles take place simultaneously including some connecting animals and humans. Investigating these transmission networks may be useful in understanding the transmission dynamics, infection risk and therefore in planning L. infantum control strategies.