Bovine leukosis virus, bovine viral diarrhea, and bovine neosporosis seroprevalence in specialized dairy herds in Antioquia-Colombia.

Enzootic bovine leukosis (EBL) is a chronic infectious disease caused by the bovine leukosis virus (BLV), a Deltaretrovirus. Bovine viral diarrhea (BVD) is an infectious disease caused by a pestivirus. Bovine neosporosis is caused by the obligate intracellular parasite Neospora caninum (Nc). These pathogens can have horizontal (postnatal) or vertical (transplacental) transmissions and affect the productive and reproductive performance of infected bovines. This work aimed to detect BLV, BVD, and Nc seroprevalence in specialized dairy cattle from the north, east, and Aburrá Valley regions of the Department of Antioquia, the highest in milk production regions in Antioquia. A total of 599 blood samples, obtained from 53 specialized dairy cattle herds, were evaluated by the ELISA test. The results revealed a seroprevalence of 41.13% for BLV (242/599), 28.48% (163/599) for Nc, and 22.7% (132/599) for BVD. Regarding the regional seroprevalence evaluation, BLV was found in 47.02% of the samples from the east, 36.87% from the north, and 46.02% from the Aburrá Valley. Nc was found in 31.03% of the samples from the east, 24.26% from the north, and 36.63% from Aburrá Valley. BVD was found in 21.62% of the samples from the east, 25.03% from the north region, and 10.39% of the samples from the Aburrá Valley. It is highlighted by these results that the north region, with the highest milk production in Antioquia, had the lowest BLV and Nc seroprevalences but the highest seroprevalence of BVD. BLV has increased in Antioquia in recent years, and as an immunosuppressive infection, opportunities for other pathogens are created by it. A significant statistical difference was found in the average prevalence of the pathogens according to the municipality, cattle breed, and region of origin of the sample. The seroprevalence of these pathogens in specialized dairy herds in Antioquia can be classified as medium-low. However, it is recommended that biosecurity practices should be maximized to avoid the spread of these pathogens due to the variability detected in the region, municipality, breed group, and herd age. The rapid and efficient diagnosis of these three pathogens through reliable methodologies will allow for the control of dissemination in dairy herds.

Effects of varying extracellular amino acid concentrations on bidirectional amino acid transport and intracellular fluxes in mammary epithelial cells.

Understanding the regulation of cellular AA uptake as protein supply changes is critical for predicting milk component yields because intracellular supplies partly regulate protein synthesis. Our objective was to evaluate cellular uptake and kinetic behavior of individual AA when cells are presented with varying extracellular AA supplies. Bovine primary mammary epithelial cells were grown to confluency and transferred to medium with an AA profile and concentration similar to that of plasma from dairy cows for 24 h. Treatments were 4 AA concentrations, 0.36, 2.30, 4.28, and 6.24 mM, which represented 16, 100, 186, and 271% of typical plasma AA concentrations, respectively, in lactating dairy cows. Twenty-four plates of cells (89.4 × 19.2 mm) were assigned to each treatment. Cells were first subjected to treatment medium enriched with 15N-labeled AA for 24 h and then incubated with treatment medium enriched with 13C-labeled AA for 0, 15, 60, 300, 900, 1,800, and 3,600 s. Intracellular free AA, intracellular protein-bound AA, and extracellular medium free AA were analyzed for concentrations and isotopic enrichment using gas chromatography-mass spectrometry. A dynamic, 12-pool model was fitted to the data for 14 AA to derive unidirectional uptake and efflux, protein turnover, transamination, oxidation, and synthesis. The derived concentration for half the maximal uptake (km) indicated no saturation of AA uptake at typical in vivo concentrations for 11 of the 14 AA. Arginine, Pro, and Val appeared to exhibit saturation kinetics. Net uptake of all essential AA except Phe was positive across treatments. Most nonessential AA exhibited negative net uptake values. Efflux of AA was quite high, with several AA exhibiting greater than 100% efflux of the respective influx. Intracellular pool turnover was rapid for most AA (e.g., 2 min for Arg), demonstrating plasticity in matching needs for protein translation to supplies. Intracellular AA concentrations increased linearly in response to treatment for most AA, whereas 9 AA exhibited quadratic responses. Amino acid uptake is responsive to varying extracellular supplies to maintain homeostasis. No saturation of uptake was evident for most AA, indicating that transporter capacity is likely not a limitation for most AA except possibly Arg, Val, and Pro in mammary epithelial cells.

An in vitro method for assessment of amino acid bidirectional transport and intracellular metabolic fluxes in mammary epithelial cells.

Understanding uptake of AA by mammary tissue as supply varies is critical for predicting milk component production. Our objective was to develop an in vitro method to quantify cellular uptake, efflux, and intracellular metabolism of individual AA that could be implemented for evaluating these factors when AA supply and profile are varied. Bovine primary mammary epithelial cells were grown to confluency and exposed to medium with an AA profile and concentration similar to lactating dairy cow plasma for 24 h. Cells were then preloaded in medium enriched with 15N-labeled AA for 24 h followed by removal of the 15N-labeled medium and incubation with medium enriched with 13C-labeled AA for 0, 15, 60, 300, 900, 1,800, and 3,600 s. Extracellular free AA and intracellular free and protein-bound AA were analyzed for concentrations and isotopic enrichment by gas chromatography-mass spectrometry. A dynamic, 12-pool model was constructed representing extracellular and intracellular free and protein-bound pools of an AA, and their respective 15N and 13C isotopes. Markov chain Monte Carlo simulation (n = 5,000) was conducted to evaluate prediction errors by deriving standard errors and posterior distributions for rate constants, fluxes, and pools. Cellular Ala influx and efflux were higher than Leu, reflecting Ala role in driving system L transport and the high capacity of sodium-dependent transport. The Ala and Leu turnover rates were 181 and 95, 580 and 857, and 74 and 157% per hour for extracellular, intracellular, and fast protein-bound pools, respectively. The intracellular and extracellular Ala to Leu ratios were quite different, meaning the blood AA profile is not the AA profile provided for protein translation. The high level of exchange and rapid turnover of pools provide a mechanism for matching the AA supplies to the precision necessary for translation. This also understates the importance of using experimental medium similar to what is observed in vivo given that some AA depend on other AA for influx (exchange driven). The average root mean squared prediction error across the isotope enrichments, pools, and concentrations was 9.7 and 14.1% for Ala and Leu, respectively, and collinearity among parameters was low, indicating adequate fit and identifiability. The described model provides insight on individual AA transport kinetics and a method for future evaluation of AA transport and intracellular metabolism when subjected to varying AA supplies.

Loss of SIRT2 leads to axonal degeneration and locomotor disability associated with redox and energy imbalance.

Sirtuin 2 (SIRT2) is a member of a family of NAD+ -dependent histone deacetylases (HDAC) that play diverse roles in cellular metabolism and especially for aging process. SIRT2 is located in the nucleus, cytoplasm, and mitochondria, is highly expressed in the central nervous system (CNS), and has been reported to regulate a variety of processes including oxidative stress, genome integrity, and myelination. However, little is known about the role of SIRT2 in the nervous system specifically during aging. Here, we show that middle-aged, 13-month-old mice lacking SIRT2 exhibit locomotor dysfunction due to axonal degeneration, which was not present in young SIRT2 mice. In addition, these Sirt2-/- mice exhibit mitochondrial depletion resulting in energy failure, and redox dyshomeostasis. Our results provide a novel link between SIRT2 and physiological aging impacting the axonal compartment of the central nervous system, while supporting a major role for SIRT2 in orchestrating its metabolic regulation. This underscores the value of SIRT2 as a therapeutic target in the most prevalent neurodegenerative diseases that undergo with axonal degeneration associated with redox and energetic dyshomeostasis.

Caracterización citogenética del pez neotropical Brycon henni (Pisces: Characidae)

Cytogenetic characterization of the neotropical fish Brycon henni (Pisces: Characidae). Brycon henni, is a native fish from Western Colombia is important as food and in sport fishing, and could be cultured in waters between 18 and 28ºC. A previous cytogenetic study in branchial cells indicated different chromosomal complements. Total blood samples were seeded in 4 ml Ham-F12 culture medium, supplemented with 0.5 ml Bovine Fetal Serum and 0.7 ml Phyitohemaglutinin M (Gibco®) during 96 hr at 28ºC; an antimitotic factor (100µl Colcemid® 10%) was added. Cells were incubated in 10 ml KCl hypotonic solution (0.56%) for 24 min at 37ºC, fixed in methanol; acetic acid (3: 1) three times, separated by centrifugations, extended drip, and the chromosomes slides were stained with 5% Giemsa (5%). The best metaphases (6 males and 8 females) were photographed. There was a diploid number of 50 chromosomes: Metacentrics (26M), Submetacentrics (16SM) and Subtelocentrics (8ST). The fundamental number is 100 arms. The evolutionary tendency is type B with no supernumerary chromosomes; a first pair of big metacentric chromosomes is characteristic for Brycon, while no chromosomic sexual heteromorfism was detected. Rev. Biol. Trop. 56 (4): 1619-1628. Epub 2008 December 12.

La sabaleta Brycon henni, es un pez nativo del occidente colombiano importante para la seguridad alimentaria de las poblaciones ribereñas y para la pesca deportiva por sus característica combativas. El desarrollarse entre los 18 y 28ºC la convierte en una especie de cultivo promisoria. Un estudio citogenético a partir de células branquiales indicó diferentes complementos cromosómicos. Este trabajo aplicó la técnica de cultivo de linfocitos a muestras de sangre de adultos. Las muestras de sangre total fueron sembradas en 4 ml de medio de cultivo Ham-F12, suplementado con 0.5 ml de Suero Fetal Bovino y 0.7 ml del mitógeno Fitohemaglutinina M (Gibco®) durante 96 hr a 28ºC; 1.5 hr antes de la cosecha agregamos 100µl de Colcemid al 10% como factor antimitótico. Cada cultivo se incubó con 10 ml de solución hipotónica KCl 0.56% por 24 min a 37ºC, para continuar con tres fijaciones sucesivas, centrifugaciones y tinción Giemsa al 5%. Las mejores metafases fueron fotografiadas en microscopio, correspondientes a 6 machos y 8 hembras, indicando un número diploide de 50 cromosomas, clasificados en Metacéntricos (26M), Submetacéntricos (16SM) y Subtelocéntricos (8ST), para un número fundamental (NF) de 100 brazos. La tendencia evolutiva hallada fue de tipo B; no se encontraron cromosomas supernumerarios pero sí un primer par de cromosomas metacéntricos grandes, considerado marcador para el género Brycon, no determinante de heteromorfismo sexual. Estos resultados coinciden con los demás bricónidos investigados, en donde se podría considerar un ancestro común con un número cromosómico básico y cariotipos simétricos.

[Cytogenetic characterization of the neotropical fish Brycon henni (Pisces: Characidae)]. / Caracterización citogenética del pez neotropical Brycon henni (Pisces: Characidae).

Brycon henni, is a native fish from Western Colombia is important as food and in sport fishing, and could be cultured in waters between 18 and 28 degrees C. A previous cytogenetic study in branchial cells indicated different chromosomal complements. Total blood samples were seeded in 4 ml Ham-F12 culture medium, supplemented with 0.5 ml Bovine Fetal Serum and 0.7 ml Phyitohemaglutinin M (Gibco) during 96 hr at 28 degrees C; an antimitotic factor (100 microl Colcemid 10%) was added. Cells were incubated in 10 ml KCl hypotonic solution (0.56%) for 24 min at 37 degrees C, fixed in methanol; acetic acid (3: 1) three times, separated by centrifugations, extended drip, and the chromosomes slides were stained with 5% Giemsa (5%). The best metaphases (6 males and 8 females) were photographed. There was a diploid number of 50 chromosomes: Metacentrics (26M), Submetacentrics (16SM) and Subtelocentrics (8ST). The fundamental number is 100 arms. The evolutionary tendency is type B with no supernumerary chromosomes; a first pair of big metacentric chromosomes is characteristic for Brycon, while no chromosomic sexual heteromorfism was detected.