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Genetic characterisation of African swine fever virus in outbreaks in Ha Nam province, Red River Delta Region of Vietnam, and activity of antimicrobial products against virus infection in contaminated feed
2020
Tran, Ha Thi Thanh | Truong, Anh Duc | Ly, Duc Viet | Vũ, Thị Hảo | Hoàng, Văn Tuấn | Nguyễn, Thị Chính | Chu, Thi Nhu | Nguyen, Thi Huyen | Pham, Ngoc Thi | Nguyen, Tinh | Yersin, Andrew G. | Dang, Hoang Vu
African swine fever (ASF) was officially reported in Vietnam in February 2019 and spread across the whole country, affecting all 63 provinces and cities. In this study, ASF virus (ASFV) VN/Pig/HaNam/2019 (VN/Pig/HN/19) strain was isolated in primary porcine alveolar macrophage (PAM) cells from a sample originating from an outbreak farm in Vietnam’s Red River Delta region. The isolate was characterised using the haemadsorption (HAD) test, real-time PCR, and sequencing. The activity of antimicrobial feed products was evaluated via a contaminated ASFV feed assay. Phylogenetic analysis of the viral p72 and EP402R genes placed VN/Pig/HN/19 in genotype II and serogroup 8 and related it closely to Eastern European and Chinese strains. Infectious titres of the virus propagated in primary PAMs were 10⁶ HAD₅₀/ml. Our study reports the activity against ASFV VN/Pig/HN/19 strain of antimicrobial Sal CURB RM E Liquid, F2 Dry and K2 Liquid. Our feed assay findings suggest that the antimicrobial RM E Liquid has a strong effect against ASFV replication. These results suggest that among the Sal CURB products, the antimicrobial RM E Liquid may have the most potential as a mitigant feed additive for ASFV infection. Therefore, further studies on the use of antimicrobial Sal CURB RM E Liquid in vivo are required. Our study demonstrates the threat of ASFV and emphasises the need to control and eradicate it in Vietnam by multiple measures.
Show more [+] Less [-]Evaluation of the effect of surgical and immunological castration of male pigs on boar taint compounds in oral fluid and fat tissue by LC-MS/MS method
2020
Woźniak-Sobczak, Barbara | Cybulski, Piotr | Jabłoński, Artur | Witek, Sebastian | Matraszek-Żuchowska, Iwona
An effective way of preventing undesirable boar taint in pork meat caused by the presence of androstenone, skatole and indole is surgical castration of piglets. This, however, arouses growing social opposition. An alternative method of inhibiting the development of unpleasant odour is immune castration. The aim of the study was to compare the effectiveness of both methods of castration for the elimination of the compounds responsible and to assess the suitability of oral fluid for pre-slaughter predictive testing for boar taint. The research material was pooled oral fluid and fat samples taken from gilts and surgically and immunologically castrated piglets. The samples were tested with a liquid chromatography– tandem mass spectrometry method developed in this research. The compounds giving rise to boar taint were found only sporadically above the accepted limits; only one sample of oral fluid contained skatole at a concentration above 200 μg L⁻¹ and one contained indole more concentrated than 100 μg L⁻¹. Indole above the limit value was also detected in one fat sample. In none of the tested samples was androstenone found. The results indicate the similar effectiveness of both methods of piglet castration on the reduction of compounds generating boar taint. The usefulness of testing oral fluid for the ante-mortem prediction of boar taint has not been fully confirmed and further investigation is needed.
Show more [+] Less [-]New method of analysis of nitrofurans and nitrofuran metabolites in different biological matrices using UHPLC-MS/MS
2018
Śniegocki, Tomasz | Giergiel, Marta | Sell, Bartosz | Posyniak, Andrzej
The major difficulty in analysis of nitrofurans in feed, feed water, and food of animal origin is that nitrofurans have low molecular weights and fast metabolism. The principal goal of this study was to prepare a procedure for the determination of nitrofurans and their metabolites by a single method in different types of feed, feed water, and food of animal origin. Two-gram samples were subjected to hydrolysis and derivatisation processes by addition of hydrochloric acid and 2-nitrobenzaldehyde. After incubation the sample was purified by solid phase extraction technique. Nitrofurans were analysed using ultra-high-pressure liquid chromatography-MS/MS (UHPLC-MS/MS). The results of validation fulfil the requirement of the confirmatory criteria according to the European Commission Decision 2002/657/EC regarding apparent recoveries (88.9%–107.3%), repeatability (2.9%–9.4%) and within-laboratory reproducibility (4.4%–10.7%). The method can be successfully applied to monitor nitrofurans and their metabolites in different matrices.
Show more [+] Less [-]In-house validation method for quantification of amoxicillin in medicated feedingstuffs with the use of HPLC-DAD technique
2020
Patyra, Ewelina | Kwiatek, Krzysztof
A high-performance liquid chromatographic–diode array detector (HPLC-DAD) method for the determination of amoxicillin in medicated feedingstuffs was developed and validated. The method was used to investigate the quality requirements of animal feedingstuffs (declared content of active substance and feed homogeneity). Two-gram samples were extracted by potassium phosphate buffer solution. Extracts were filtered and directly analysed by HPLC-DAD without further clean-up. Amoxicillin was separated by acetonitrile and 0.01M phosphate buffer (pH 5.0) on a Phenomenex Luna C18 column. This method provided average recoveries of 76.1 to 81.6% with coefficients of variation (CV, %) for repeatability and reproducibility in the ranges of 3.7–7.2% and 5.3–7.6%, respectively. The limit of detection was 51.2 mg/kg and limit of quantification was 103.0 mg/kg. The method was successfully validated and proved to be efficient, precise, and useful for quantification of amoxicillin in medicated feedingstuffs.
Show more [+] Less [-]Effects of lung site and fluid volume on results of bronchoalveolar lavage fluid analysis in horses
1992
Sweeney, C.R. | Rossier, Y. | Ziemer, E.L. | Lindborg, S.
Bronchoalveolar lavage (BAL) fluid was analyzed in healthy horses, using different lavage fluid volumes and lung sites. The only significant difference in the cellular composition of BAL fluid between the right and left lungs was the mast cell numbers, which were significantly higher in the left lung. Total cell count ranged from 34 to 330 cells/microliter for the right lung and 43 to 330 cells/microliter for the left lung. Percentage of neutrophils ranged from 1 to 7% in the right lung and 1 to 5% in the left lung. The small-volume (50 ml) lavage had a greater percentage of neutrophils and a lesser percentage of mast cells in the large-volume (350 ml) lavage. Statistical difference in the composition of BAL fluid recovered was not detected between the 3 sequential 100-ml lavages and a single 300-ml lavage, except that macrophages were significantly higher in the 3 sequential 100-ml lavages. Values for BAL fluid analysis in healthy horses have varied considerably and this variation is from a failure to adhere to any standard technique for volume of fluid infused.
Show more [+] Less [-]Gastric emptying of a radionuclide-labeled test meal after surgical correction of gastric dilatation-volvulus in dogs
1989
Sluijs, F.J. van | Brom, W.E. van der
Gastric emptying of a radionuclide-labeled test meal was studied in 10 dogs that had been treated surgically for gastric dilatation-volvulus and in 10 clinically normal (control) dogs. There were no significant differences between the gastric emptying rates and patterns in treated and in control dogs. Thus, there are no indications that gastric emptying is delayed in dogs that have recovered from gastric dilatation-volvulus, and there is no reason for pyloric surgery in dogs with this condition.
Show more [+] Less [-]Survival of Mycobacterium bovis during forage ensiling
2019
Grooms, Dan | Bolin, Steven R. | Plastow, Jessica L. | Lim, Ailam | Hattey, Joseph | Durst, Phillip T. | Rust, Steven R. | Allen, Michael S. | Buskirk, Daniel D. | Smith, Richard W.
OBJECTIVE To determine whether Mycobacterium bovis remains viable in ensiled forages. SAMPLE Alfalfa, mixed mostly grass, and corn silages. PROCEDURES For each of 10 sampling days, six 250-g replicate samples of each feedstuff were created and placed in a film pouch that could be vacuum sealed to simulate the ensiling process. Within each set of replicate samples, 4 were inoculated with 10 mL of mycobacterial liquid culture medium containing viable M bovis and 2 were inoculated with 10 mL of sterile mycobacterial liquid culture medium (controls) on day 0. Pouches were vacuum sealed and stored in the dark at room temperature. On the designated sampling day, 1 control pouch was submitted for forage analysis, and the other pouches were opened, and forage samples were obtained for M bovis culture and analysis with a PCR assay immediately and 24 hours later. RESULTS None of the control samples had positive M bovis culture or PCR assay results. Among M bovis-inoculated samples, the organism was not cultured from alfalfa and corn silage for > 2 days but was cultured from mixed mostly grass silage for 28 days after inoculation and ensiling initiation. Mycobacterium bovis DNA was detected by PCR assay in samples of all 3 feedstuffs throughout the 112-day observation period. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that properly ensiled forages would be an unlikely source for M bovis transmission to cattle. Further research is necessary to determine whether ensiling kills M bovis or forces it to become dormant and, if the latter, elucidate the conditions that cause it to revert to an infectious state.
Show more [+] Less [-]Evaluation of vaporized hydrogen peroxide sterilization on the in vitro efficacy of meropenem-impregnated polymethyl methacrylate beads
2019
Druham, Myra E. | Elfenbein, Johanna R.
OBJECTIVE To evaluate the effects of vaporized hydrogen peroxide (VHP) sterilization on the in vitro antimicrobial efficacy of meropenem-impregnated polymethyl methacrylate (M-PMMA) beads. SAMPLE 6-mm-diameter polymethyl methacrylate beads that were or were not impregnated with meropenem. PROCEDURES Meropenem-free polymethyl methacrylate and M-PMMA beads were sterilized by use of an autoclave or VHP or remained unsterilized. To determine the antimicrobial efficacy of each bead-sterilization combination (treatment), Mueller-Hinton agar plates were inoculated with 1 of 6 common equine pathogens, and 1 bead from each treatment was applied to a sixth of each plate. The zone of bacterial inhibition for each treatment was measured after 24 hours. To estimate the duration of antimicrobial elution into a solid or liquid medium, 1 bead from each treatment was transferred every 24 hours to a new Staphylococcus aureus–inoculated agar plate or a tube with PBS solution, and an aliquot of the eluent from each tube was then applied to a paper disc on an S aureus–inoculated agar plate. All agar plates were incubated for 24 hours, and the zone of bacterial inhibition was measured for each treatment. RESULTS In vitro antimicrobial efficacy of M-PMMA beads was retained following VHP sterilization. The duration of antimicrobial elution in solid and liquid media did not differ significantly between unsterilized and VHP-sterilized M-PMMA beads. CONCLUSIONS AND CLINICAL RELEVANCE Results indicated that M-PMMA beads retained in vitro antimicrobial activity and eluted the drug for up to 2 weeks after VHP sterilization.
Show more [+] Less [-]Effects of a nutrient-enriched water on water intake and indices of hydration in healthy domestic cats fed a dry kibble diet
2018
Zanghi, Brian M. | Gerheart, Lynn | Gardner, Cari L.
OBJECTIVE To evaluate the effects of drinking nutrient-enriched water (NW) on water intake and indices of hydration in healthy domestic cats fed a dry kibble diet ad libitum. ANIMALS 18 domestic shorthair cats. PROCEDURES Group-housed cats were assigned to tap water (TW; n = 9) or NW (9) groups. All cats received TW at baseline (days −7 to −1). No changes were made to the food-water regimen for the TW group. The NW group received NW instead of TW from days 0 through 10, then received TW and NW in separate bowls (days 11 through 56). Food intake was measured through day 10; liquid consumed by drinking was measured throughout the study. Blood and urine samples were collected at predetermined times for analyses; 48-hour urine collection (days 28 through 30 or 31 through 33) was performed to assess output volume and aid endogenous creatinine-based glomerular filtration rate (GFR) determination. Data were analyzed with linear mixed-effects models. RESULTS Baseline TW and calorie intake were similar between groups. The NW treatment was significantly associated with increased liquid consumption during the treatment phase. Mean urine output was significantly higher in the NW group (15.2 mL/kg/d) than in the TW group (10.3 mL/kg/d). Mean GFR (1.75 vs 1.87 mL/min/kg, respectively) did not differ between groups. Effects of treatment and time were each significant for urine specific gravity and osmolality and urine creatinine, phosphate, and urea nitrogen concentrations, with lower values for the NW group. CONCLUSIONS AND CLINICAL RELEVANCE Results suggested that consumption of the NW can increase liquid intake and improve measures of hydration in healthy cats. These effects may offer health benefits to some cats in need of greater water consumption.
Show more [+] Less [-]Effect of hypertonic vs isotonic saline solution on responses to sublethal Escherichia coli endotoxemia in horses
1990
Bertone, J.J. | Gossett, K.A. | Shoemaker, K.E. | Bertone, A.L. | Schneiter, H.L.
Cardiovascular responses to sublethal endotoxin infusion (Escherichia coli, 50 micrograms/ml in lactated Ringer solution at 100 ml/h until pulmonary arterial pressure increased by 10 mm of Hg) were measured 2 times in 5 standing horses. In a 2-period crossover experimental design, horses were either administered hypertonic (2,400 mosm/kg of body weight, IV) or isotonic (300 mosm/kg, IV) NaCl solution after endotoxin challenges. Each solution was administered at a dose of 5 ml/kg (infusion rate, 80 ml/min). Complete data sets (mean arterial, central venous, and pulmonary arterial pressures, pulmonary arterial blood temperature, cardiac output, total peripheral vascular resistance, heart rate, plasma osmolality, plasma concentration of Na, K, Cl, and total protein, blood lactate concentration, and PCV) were collected at 0 (baseline, before endotoxin infusion), 0.25, 1, 1.5, 2, 2.5, 3, 3.5, 4, and 4.5 hours after initiation of the endotoxin infusion. Blood constituents alone were measured at 0.5 hour and cardiovascular variables alone were evaluated at 0.75 hour. By 0.25 hour, endotoxin infusion was completed, a data set was collected, and saline infusion was initiated. By 0.75 hour, saline solutions had been completely administered. Mean (+/- SEM) cardiac output decreased (99.76 +/- 3.66 to 72.7 +/- 2.35 ml/min/kg) and total peripheral resistance (1.0 +/- 0.047 to 1.37 +/- 0.049 mm of Hg/ml/min/kg) and pulmonary arterial pressure (33.4 +/- 0.86 to 58.3 +/- 1.18 mm of Hg) increased for both trials by 0.25 hour after initiation of the endotoxin infusion and prior to fluid administration. For the remainder of the protocol, cardiac output was increased and total peripheral resistance was decreased during the hypertonic, compared with the isotonic, saline trial. Cardiac output was decreased and total peripheral resistance was increased during the isotonic saline trial, compared with baseline values. Both trials were associated with increased blood lactate concentration, but lactate values during the isotonic saline trial were greater and remained increased above baseline values for a longer period (4 hours) than during the hypertonic saline trial (2.5 hours). It was concluded for this model of endotoxemia, that IV administered hypertonic saline solution was associated with more-desirable cardiovascular and metabolic responses than was an equal volume of isotonic saline solution.
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