High genetic diversity and zoonotic potential of Staphylococcus aureus strains recovered from bovine intramammary infections in Colombians dairy herds.

Genotyping of Staphylococcus aureus isolated from mastitis has become a fundamental tool to understand its complex epidemiology and to evaluate spillover events. The aim of this study was to describe the frequency of genotypes of the S. aureus strains isolated from intramammary infections by spa typing technique, and to evaluate the association between genotypes and the ability to form biofilm under in vitro conditions. Sixty-six strains of S. aureus recovered from bovines intramammary infections on 56 dairy herds located in 14 municipalities of the department of Antioquia were characterized. The majority of strains (65/66) were isolated from milk samples collected from dairy cows with subclinical intramammary infections. Nineteen different spa types were found in this study, t521 (19.70%), t267 (15.15%), and t605 (12.12%) being the most frequent. The strains from the t605 spa type showed the highest biofilm production. The high frequency of spa types with zoonotic potential found in this study, identified cattle as an important reservoir of theses clones for people in close proximity, such as milkers and consumers of unpasteurized dairy products.

Genotypic and phenotypic characterization of biofilm production by Staphylococcus aureus strains isolated from bovine intramammary infections in Colombian dairy farms.

The ability of Staphylococcus aureus to form biofilms is an important virulence factor because this has been associated with persistent bovine intramammary infections. Different mechanisms of biofilm formation have been described in S. aureus; however, the process has been found to be mainly driven by the ica and bap genes. The presence of the ica and bap genes, as well as the biofilm formation in vitro were evaluated in 229 S. aureus strains isolated from bovine milk collected from different regions of Department of Antioquia, Colombia. Three different genotypes grouped into three separate clusters were identified from in vitro assays. Genotype 1 (ica positive and bap negative) was the most prevalent (78.17%), followed by genotype 2 (ica and bap positive) (12.66%) and genotype 0 (ica and bap negative) (9.17%). Biofilm formation was observed in 81.26% of the strains from which 100% of genotype 2 isolates showed biofilm formation. The biofilms formed by genotype 2 isolates were also found to have the highest optical density (>2.4). These results showed that most of the S. aureus strains were capable of biofilm formation, suggesting the virulence potential particularly in bap-positive strains.

Characterization and classification of the compositional quality of milk from river buffaloes (Bubalus bubalis) and cows (Bos spp.) in Colombia / Caracterización y clasificación de la calidad composicional de la leche de búfalo (Bubalus bubalis) y de vaca (Bos spp.) en Colombia

ABSTRACT The aim of this study was to classify and characterize the compositional quality of milk from river buffaloes (Bubalus bubalis) and cows (Bos spp.) in Colombia based on the fat, protein, and total solid (TS) contents. Using a hierarchical procedure, data on milk from river buffaloes (n = 7,726) and cows (n = 49,330) were filtered and subjected to cluster analysis in order to generate three groups: Normal (N), High quality (HQ) and Poor Quality (PQ). The categorized database was then randomly separated into two sets (training and validation) and a discriminant analysis was applied. In total, 37.3% of river buffalo milk samples were classified as N (6.80% fat, 4.34% protein, and 16.80% TS), 13% as HQ (9.41% fat, 4.93% protein, and 19.50% TS), and 43.7% as PQ (3.95% fat, 3.92% protein, and 13.7% TS). In contrast, 41.8% of cow milk samples were classified as N (3.64% fat, 3.37% protein, 12.42% TS), 41.2% as PQ (2.71% fat, 3.08% protein, and 10.6% TS), and 16.9% as HQ (5.46% fat, 4.01% protein, and 14.82% TS). The discriminant models for both river buffalo and cow milk were able to classify milk in the N and PQ groups with >90% accuracy, and that in the HQ group with >85% accuracy.

RESUMEN El objetivo de este estudio fue caracterizar y clasificar la calidad composicional de la leche de búfala (Bubalus bubalis) y de vaca (Bos spp.) en Colombia con base en los contenidos de grasa, proteína y sólidos totales. Mediante un procedimiento jerárquico los datos de leche de búfalos de agua (n = 7,726) y vacas (n = 49,330) se filtraron y se sometieron a análisis de conglomerados para generar tres grupos: Normal (N), Alta calidad (HQ) y Calidad deficiente (PQ). La base de datos categorizada se separó aleatoriamente en dos conjuntos (entrenamiento y validación) y se aplicó un análisis discriminante. En total, 37,3% de las muestras de leche de búfalo de agua se clasificaron como N (6,80% de grasa, 4,34% de proteína y 16,80% de TS); 13% como HQ (9,41% de grasa, 4,93% de proteína y 19,50% de TS) y 43,7 % como PQ (3,95% de grasa, 3,92% de proteína y 13,7% de TS). En contraste, el 41,8% de las muestras de leche de vaca se clasificaron como N (3,64% grasa, 3,37% proteína, 12,42% TS); 16,9% como HQ (5,46% de grasa, 4,01% de proteína y 14,82% de TS) y 41,2% como PQ (2,71% grasa, 3,08% proteína y 10,6% TS). Los modelos discriminantes para el búfalo de agua y la leche de vaca fueron capaces de clasificar la leche en los grupos N y PQ con una precisión >90% y en el grupo HQ con >85% de precisión.

Efecto de la pluviosidad y el brillo solar sobre la producción y características del semen en el pez Brycon henni (Pisces: Characidae) / Effect of pluviosity and sun shine on sperm production and seminal characteristics of the fish Brycon henni (Pisces: Characidae)

In Colombia the fish Brycon henni is a protected endemic species. It inhabits water bodies in coffee producing areas (700-1900 ma.s.l.; 4º35’56’’ N -74º04’51’’ W; 18-28°C). Insufficient knowledge of its basic biology and behavior prevent the commercial culture of this promising fish. We studied the production and sperm physiology of captive males. Along a year 20 samples were taken from each of 10 males. The sample was obtained by abdominal cefalo-caudal massage and transported to the laboratory at 4°C. Except for September and October (maximum rainfall),sperm was always obtained in at least 50% of the males. Color, osmolality and pH were similar in all the samples. Volume, concentration, viability, motility and activation time were variable: sunshine had a positive effect on volume (Spearman p<0.05) and on sperm concentration (Spearman p<0.05) while pluviosity had a negative effect on volume and viability. The proportion of ions (Na,K,Mg,Ca)was constant along the year; Na being 10-fold higher than K and 100-fold higher than Mg and Ca; however the absolute concentration of all ions was slightly higher in April and in July (with no apparent relation with the other variables analyzed).

El pez Brycon henni es una especie endémica protegida por la legislación colombiana, que habita cuerpos de agua de zonas cafeteras (700-1900 m. s. n. m),comprendidas entre los 4º35’56’’ N y 74º04’51’’ W, con temperaturas que oscilan entre los 18 y los 28ºC. A pesar de las características promisorias de esta especie, su reproducción en cautiverio a nivel comercial no ha sido posible por falta de conocimientos básicos de su biología y comportamiento. El objetivo del presente trabajo fue caracterizar la producción y la fisiología espermática de machos en cautiverio. A lo largo de un año se tomaron 20 muestras de cada uno de 10 ejemplares. El semen se obtuvo mediante masaje abdominal cráneo-caudal y se transportó a 4ºC para su análisis en el laboratorio. Con excepción de septiembre y octubre que fueron los meses más lluviosos, siempre se obtuvo semen de al menos el 50% de los animales. El color, la osmolalidad y el pH fueron similares en todas las muestras a lo largo del año. El volumen, la concentración, la viabilidad, la movilidad y el tiempo de activación fueron variables: El efecto del brillo solar fue positivo sobre el volumen (Spearman p<0.05) y sobre la concentración espermática (Spearman p<0.05), mientras que la pluviosidad tuvo un efecto negativo sobre el volumen y la viabilidad. La proporción entre los iones (Na, K, Mg, Ca) se mantuvo constante durante todo el año: el sodio fue el mayor, seguido por el potasio, y por el magnesio y el calcio, 10 y 100 veces menos, respectivamente; sin embargo la concentración absoluta de estos iones aumentó durante los meses de abril y julio sin una relación aparente con las otras variables analizadas.

Mitochondrial activity, distribution and segregation in bovine oocytes and in embryos produced in vitro.

Contents Bovine oocytes and embryos produced in vitro were studied to determine the mitochondrial pattern of distribution, segregation and activity using DIOC 6 and Jc-1 fluorescence. The highest fluorescence level observed in mature oocytes was taken as 100% activity and six activity levels were estimated as follows: (1) 0%, (2) 1-15%, (3) 16-30%, (4) 31-50%, (5) 51-75% and (6) 76-100%. Three patterns of mitochondrial distribution were found: (1) diffused throughout the cytoplasm in oocytes and embryos, (2) pericytoplasmic in oocytes and embryos, and (3) perinuclear only in embryos. The segregation of mitochondria in blastomeres showed two distinct patterns: (1) symmetrical with an even mitochondrial population, and (2) asymmetrical with different numbers of mitochondria in each blastomere. In immature oocytes, mitochondrial activity was very low and the distribution was diffuse or negligible, while in mature oocytes the activity was high and the distribution was diffuse or pericytoplasmic. Competent embryos up to the 16-cell stage showed intermediate levels of activity (16-50%) but activity decreased thereafter up to the blastocyst stage. Non-competent embryos showed low levels of activity (1-15%) at all stages. These results suggest that mitochondria might play an important role during early development and that a minimum threshold of activity regulates the potential competence for reaching the blastocyst stage.

[Effect of pluviosity and sun shine on sperm production and seminal characteristics of the fish Brycon henni (Pisces: Characidae)]. / Efecto de la pluviosidad y el brillo solar sobre la producción y características del semen en el pez Brycon henni (Pisces: Characidae).

Effect of pluviosity and sun shine on sperm production and seminal characteristics of the fish Brycon henni (Pisces: Characidae). In Colombia the fish Brycon henni is a protected endemic species. It inhabits water bodies in coffee producing areas (700-1900 ma.s.l.; 4 degrees 35'56" N - 74 degrees 04'51" W; 18-28 degrees C). Insufficient knowledge of its basic biology and behavior prevent the commercial culture of this promising fish. We studied the production and sperm physiology of captive males. Along a year 20 samples were taken from each of 10 males. The sample was obtained by abdominal cefalo-caudal massage and transported to the laboratory at 4 degrees C. Except for September and October (maximum rainfall), sperm was always obtained in at least 50% of the males. Color, osmolality and pH were similar in all the samples. Volume, concentration, viability, motility and activation time were variable: sunshine had a positive effect on volume (Spearman p < 0.05) and on sperm concentration (Spearman p < 0.05) while pluviosity had a negative effect on volume and viability. The proportion of ions (Na, K, Mg, Ca) was constant along the year; Na being 10-fold higher than K and 100-fold higher than Mg and Ca; however the absolute concentration of all ions was slightly higher in April and in July (with no apparent relation with the other variables analyzed).

Follicular dynamics during postpartum anestrus and the first estrous cycle in suckled or non-suckled Brahman (Bos indicus) cows.

Brahman (Bos indicus) cows, were selected at 28+/-10 days after calving and analyzed by real time rectal ultrasonography three times a week, in order to evaluate and compare follicular and corpus luteum development during postpartum (PP) anestrus and the first PP estrous cycle under sylvopastoril conditions. Suckling (S, n=11) or non-suckling (NS, n=5) cows were evaluated in a zone of tropical dry forest (450m of altitude, mean temperature=27 degrees C, annual rainfall=1000mm). Estrous detection was performed twice daily by direct observation. Progesterone was quantified using RIA. From 28+/-10 days postcalving to resumption of estrous cycles, there were no differences (P>0.05) between NS and S cows for diameter of the dominant or first subordinate follicle, follicular growth rate, or interdominance interval. Silent ovulation, corpus luteum formation and subsequent progesterone concentrations ranging from 0.3 to 9. 7ng/ml, were found in both groups. The first calving to ovulation and calving to standing estrus intervals were shorter (P<0.01) in NS (34.8+/-5.81 and 41.2+/-9.03 days) than in S (65+/-4.82 and 81+/-6. 21 days) cows. Follicular development and progesterone concentrations during the first PP estrous cycle did not differ (P>0. 05) between NS and S cows. These results suggest that Brahman cows could have an early PP resumption of follicular recruitment if fed under sylvopastoril system conditions. However, non-suckled cows did have an earlier standing estrus and ovulation than did suckled cows.

Perfil espermático de un grupo de pacientes y de voluntarios en Medellín-Colombia / Sperm profile of a group of patients and voluntaries en Medellin-Colombia

Se describen los parámetros espermáticos de 333 individuos atendidos en nuestro Programa de Reproducción: 251 pacientes en estudio de pareja infértil y 82 voluntarios. Se presentan los valores de la mediana para los diferentes parámetros y el porcentaje de individuos con valores por debajo de los establecidos por la OMS. El mayor número de individuos con alteraciones en el espermograma, se encontró en el grupo de pacientes. La motilidad fue el parámetro más afectado en ambos grupos

Ovarian follicular dynamics in suckled zebu (Bos indicus) cows monitored by real time ultrasonography.

The pattern of follicular growth was studied in 17 suckled zebu cows with average body condition and under extensive management in a tropical environment (23 degrees C, 78% humidity; 2200 mm annual rainfall; 1000 m altitude). The study covered the period from parturition to weaning at 12 months postpartum (PP). Data were collected by transrectal ultrasonography (7.5 MHz) at 48 h intervals, and progesterone (P4) measurements were performed by RIA. The sequential development of ovarian follicles greater than 4 mm was followed until regression or ovulation. Ovarian activity as characterized by growth and regression of follicles of 4 to 6 mm, with sporadic dominance, and a long interdominance interval was observed in every cow and from as early as 26 +/- 2 days PP. This follicular pattern was highly variable during the first 6 months: cows presented 2 to 20 follicular waves (FW) in which a dominant follicle (DF) grew to 8 +/- 1 mm with daily growth rates of 1.1 +/- 0.5 mm/day. The duration of dominance varied from 2 to 8 days and the interdominance time interval was 0 (overlapped waves) to 60 days. Neither behavioural oestrus nor ovulation was observed during this period. From 6 to 12 months PP, cows presented 7 to 20 FW, some with ovulation and/or corpora lutea (CL) formation. The ovulation was preceded by oestrus in some cases (43%). The mean (+/- sem) diameter of DF was 9 +/- 2.7 mm, their mean growth rate 1.4 +/- 0.2 mm/day, their duration of dominance was 2 to 8 days and the interdominance interval was 0 to 14 days. Progesterone concentrations (P4) from 1.0 to 13 ng/ml were found when a CL was present. Once cyclicity re-commenced at 217 to 278 days PP, the cows presented either normal (21 +/- 3 days), short (10 +/- 2 days), or long (50 +/- 4 days) cycles. The resumption of cyclicity was characterized by an increased frequency of emerging follicular waves. Under the conditions of this study, the suckled Bos indicus cows re-commenced ovarian follicular activity as early as described in B. taurus breeds, but the establishment of cyclicity was substantially later. These data add further to the panorama of postpartum reproductive physiology in tropical cattle.

Oocyte quality in small antral follicles in the presence or absence of a large dominant follicle in cattle.

The aim of these experiments was to determine whether the presence of a dominant follicle affects the developmental competence of oocytes from small antral follicles of cattle. In Expt 1, oocytes or follicular fluid samples were collected from follicles 2-7 mm in diameter before (day 3 of the oestrous cycle; n = 4) or after (days 6 and 7 of the oestrous cycle; n = 6) emergence of the first wave dominant follicle (verified by rectal ultrasonography). Five to ten follicles were aspirated for the determination of individual follicular fluid concentrations of oestradiol, progesterone, testosterone and dimeric inhibin; oocytes from the remaining follicles from each cow were pooled and developmental capacity assessed by in vitro fertilization and maturation. In Expt 2, ovaries containing a young corpus luteum and with or without a large oestrogen-active follicle were collected from the abattoir. Follicular aspirates from small follicles in each pair of ovaries were pooled, and oocyte quality and steroid concentrations were determined. In Expt 1, small follicles obtained before emergence of the dominant follicle contained significantly more oestradiol than they did after emergence (19.5 +/- 1.5 versus 0.7 +/- 1.1 ng ml-1, respectively; P < 0.05), but there were no significant differences in concentrations of progesterone, testosterone or dimeric inhibin. The percentage of blastocysts obtained from oocytes collected before (12.1 +/- 9.0) or after emergence (11.8 +/- 7.0) of the dominant follicle did not differ significantly (P > 0.05). In Expt 2, follicular steroid concentrations did not differ in small follicles taken in the presence versus the absence of a large oestrogen-active follicle, and there were no differences in the developmental capacity of the oocytes. There were significant negative correlations between follicular oestradiol concentration and the percentage of blastocysts formed from two-cell (r = -0.90; P < 0.01) and eight-cell embryos (r = -0.65; P < 0.05). These data suggest that in cattle the developmental competence of oocytes from small antral follicles is not adversely affected by the presence of a dominant follicle.

Recovery rate and quality of embryos collected from suckled cows and beef heifers after superovulation with PMSG.

Recovery rate and embryo quality were investigated in beef heifers and suckled cows following superovulation induced by 2000 IU pregnant mare serum (PMSG) combined with different methods of estrus cycle synchronization (Norgestomet, Prid, Dinolytic, Norgestomet combined with Dinolytic). Genital tracts were flushed upon slaughter with Dulbecco's medium 6.5 to 7.5 days after insemination. Of the heifers, 42 out of 43 responded to treatment. The mean embryo recovery rate, based on the number of corpora lutea, was only 14.8%. Of the 83 embryos recovered, 54.2% had developed to the expected stage and only 40% appeared normal. Of the adult cows, 55 out 58 responded with an embryo recovery rate of 39.5%. Of the 149 embryos recovered, 48.9% had developed to the expected stage and 67.1% of these appeared normal. In both heifers and adult cows, the different methods of estrus synchronization produced no significant differences in recovery rate or embryo quality.