Genome-wide association study of periweaning failure-to-thrive syndrome (PFTS) in pigs.

Porcine periweaning-failure-to-thrive syndrome (PFTS) is a condition that affects newly weaned piglets. It is characterised by a progressive debilitation leading to death, in the absence of infectious, nutritional, management or environmental factors. In this study, we present the first report of PFTS in South America and the results of a genome-wide association study to identify the genetic markers associated with the appearance of this condition in a crossbred swine population. Four chromosomal regions were associated with PFTS predisposition, one located on SSCX, one on SSC8, and the two other regions on SSC14. Regions on SSC8 and SSC14 harbour important functional candidate genes involved in human depression and might have an important role in PFTS. Our findings contribute to the increasing knowledge about this syndrome, which has been investigated since 2007, and to the identification of the aetiology of this disease.

Quantitative trait loci associated with chemical composition of the chicken carcass.

Major objectives of the poultry industry are to increase meat production and to reduce carcass fatness, mainly abdominal fat. Information on growth performance and carcass composition are important for the selection of leaner meat chickens. To enhance our understanding of the genetic architecture underlying the chemical composition of chicken carcasses, an F(2) population developed from a broiler × layer cross was used to map quantitative trait loci (QTL) affecting protein, fat, water and ash contents in chicken carcasses. Two genetic models were applied in the QTL analysis: the line-cross and the half-sib models, both using the regression interval mapping method. Six significant and five suggestive QTL were mapped in the line-cross analysis, and four significant and six suggestive QTL were mapped in the half-sib analysis. A total of eleven QTL were mapped for fat (ether extract), five for protein, four for ash and one for water contents in the carcass using both analyses. No study to date has reported QTL for carcass chemical composition in chickens. Some QTL mapped here for carcass fat content match, as expected, QTL regions previously associated with abdominal fat in the same or in different populations, and novel QTL for protein, ash and water contents in the carcass are presented here. The results described here also reinforce the need for fine mapping and to perform multi-trait analyses to better understand the genetic architecture of these traits.

Porcine circovirus type 2 (PCV2) vaccination is effective in reducing disease and PCV2 shedding in semen of boars concurrently infected with PCV2 and Mycoplasma hyopneumoniae.

The objectives were to determine whether the amount of porcine circovirus type 2 (PCV2) shed in semen increased in boars experimentally coinfected with Mycoplasma hyopneumoniae (MHYO), and whether PCV2 vaccination of boars prior to PCV2 exposure reduced PCV2 viremia and virus shedding in semen. Twelve specific-pathogen-free PCV2- and MHYO-naïve boars were randomly and equally assigned to one of four groups. Six boars were vaccinated against PCV2 (VAC) on Day 0; three PCV2 vaccinated and three non-vaccinated boars were inoculated with MHYO on Day 21, and all boars were challenged with PCV2 on Day 35. The four treatment groups included PCV2-Infected (I), VAC-PCV2-I, MHYO-PCV2-Coinfected (CoI), and VAC-MHYO-PCV2-CoI. Semen, blood swabs, feces, and serum samples were collected weekly until Day 70. All vaccinated boars had seroconverted to PCV2 by Day 35. Between Days 28 and 35, MHYO boars developed moderate respiratory disease, characterized by coughing, respiratory distress, mucopurulent nasal discharge and loss of body condition. One MHYO-PCV2-CoI boar died on Day 50. Boars in the PCV2-I and MHYO-PCV2-CoI groups had significantly higher PCV2 DNA loads in blood swabs than the remaining boars. Moreover, PCV2 vaccination significantly reduced the incidence and amount of PCV2 shedding in semen and feces. In summary, although concurrent MHYO infection did not influence PCV2 shedding patterns, coinfection of boars with PCV2 and MHYO resulted in severe clinical disease and viral shedding was significantly decreased by PCV2 vaccination.

QTL for percentage of carcass and carcass parts in a broiler x layer cross.

An F2 experimental population, developed from a broiler layer cross, was used in a genome scan of QTL for percentage of carcass, carcass parts, shank and head. Up to 649 F2 chickens from four paternal half-sib families were genotyped with 128 genetic markers covering 22 linkage groups. Total map length was 2630 cM, covering approximately 63% of the genome. QTL interval mapping using regression methods was applied to line-cross and half-sib models. Under the line-cross model, 12 genome-wide significant QTL and 17 suggestive linkages for percentages of carcass parts, shank and head were mapped to 13 linkage groups (GGA1, 2, 3, 4, 5, 7, 8, 9, 11, 12, 14, 18 and 27). Under the paternal half-sib model, six genome-wide significant QTL and 18 suggestive linkages for percentages of carcass parts, shank and head were detected on nine chicken linkage groups (GGA1, 2, 3, 4, 5, 12, 14, 15 and 27), seven of which seemed to corroborate positions revealed by the previous model. Overall, three novel QTL of importance to the broiler industry were mapped (one significant for shank% on GGA3 and two suggestive for carcass and breast percentages on GGA14 and drums and thighs percentage on GGA15). One novel QTL for wings% was mapped to GGA3, six novel QTL (GGA1, 3, 7, 8, 9 and 27) and suggestive linkages (GGA2, 4, and 5) were mapped for head%, and suggestive linkages were identified for back% on GGA2, 11 and 12. In addition, many of the QTL mapped in this study confirmed QTL previously reported in other populations.

Mapping QTLs on chicken chromosome 1 for performance and carcass traits in a broiler x layer cross.

With the objective of mapping quantitative trait loci (QTLs) for performance and carcass traits, an F2 chicken population was developed by crossing broiler and layer lines. A total of 2063 F2 chicks in 21 full-sib families were reared as broilers and slaughtered at 42 days of age. Seventeen performance and carcass traits were measured. Parental F(0) and F1 individuals were genotyped with 80 microsatellites from chicken chromosome 1 to select informative markers. Thirty-three informative markers were used for selective genotyping of F2 individuals with extreme phenotypes for body weight at 42 days of age (BW42). Based on the regions identified by selective genotyping, seven full-sib families (649 F2 chicks) were genotyped with 26 markers. Quantitative trait loci affecting body weight, feed intake, carcass weight, drums and thighs weight and abdominal fat weight were mapped to regions already identified in other populations. Quantitative trait loci for weights of gizzard, liver, lungs, heart and feet, as well as length of intestine, not previously described in the literature were mapped on chromosome 1. This F2 population can be used to identify novel QTLs and constitutes a new resource for studies of genes related to growth and carcass traits in poultry.

Reproductive characteristics of grass-fed, luteinizing hormone-releasing hormone-immunocastrated Bos indicus bulls.

Two field trials were conducted in Brazil to evaluate LHRH immunocastration of Bos indicus bulls (d 0 = 2 yr of age). In Study I, 72 bulls were assigned randomly to one of three treatment groups: LHRH0-immunized, castrated, and intact. Immunized animals (n = 25) received a primary and two booster injections of ovalbumin-LHRH-7 and thioredoxin-LHRH-7 fusion proteins on d 0, 141, and 287. Twenty-three bulls were surgically castrated on d 141, and 24 served as intact controls. All animals were slaughtered on d 385, at approximately 3 yr of age. In Study II, 216 bulls were assigned randomly to the same three treatments as in Study I; however, because of a drought in the area, bulls were kept on pasture an additional year, and a fourth treatment was added, in which one-half the LHRH-immunized bulls received an additional booster on d 639 (fourth immunization). All animals in Study II were slaughtered on d 741 (4 yr of age). Luteinizing hormone-releasing hormone antibodies increased following each immunization for immunized bulls, but they were not detectable in castrate or intact animals in either study. Consequently, scrotal circumference was suppressed in immunized bulls compared with intact controls in both studies. By d 287, serum concentrations of testosterone in LHRH-immunized bulls were decreased compared with intact controls (P < 0.01). In both studies, testes and epididymal weights at slaughter were greater (P < 0.01) for intact (500 +/- 17 and 60 +/- 2 g, respectively) than for immunized bulls (173 +/- 22 and 26 +/- 2 g, respectively) and fourth immunization bulls (78 +/- 23 and 20 +/- 2 g, respectively; Study II). At the end of each study, BW was greater (P < 0.01) for intact bulls than for castrated and LHRH-immunized animals. In these two studies, the efficacy of the LHRH fusion proteins to induce an effect similar to that of surgical castration was considered 92 and 93%, respectively. These data support the concept that immunocastration of bulls at 2 yr of age was successful and that it has practical application as a tool for producing grass-fattened bulls in Brazil.

Regulation of frequency of luteinizing hormone pulses by magnitude of acute change in circulating concentration of progesterone of female cattle.

The hypothesis was the greater the magnitude of acute increase in circulating concentration of progesterone of female cattle, the greater the acute inhibitory effect on frequency of pulsatile LH release. From Day 0 to 4 of the treatment period, females without functional corpora lutea were treated with varying doses of progesterone to result in varying concentrations of progesterone within the typical physiological range in blood. From Day 4 to 7, cattle were treated with a single dose of progesterone to achieve a similar circulating concentration of progesterone among all females in the study. Therefore, from Day 0 to 4 relative to Day 4 to 7 of the treatment period, females had a: (1) large (3.1 ng/ml), (2) moderate (2.5 ng/ml), or (3) small (0.5 ng/ml) increase in concentration of progesterone in blood. Frequency of LH pulses was greater (P <0.10) in females with the greatest magnitude of change in concentration of progesterone during the first 24 h following the change in concentration as compared with females with the moderate or small of change in concentration of progesterone suggesting our working hypothesis should be rejected. The greater the magnitude of acute change in concentration of progesterone, however, the longer time required for re-initiation of release of LH pulses of the amplitude of pulses that preceded the change in concentration of progesterone.

Growth and carcass characteristics of pasture fed LHRH immunocastrated, castrated and intact Bos indicus bulls.

The effectiveness of a luteinizing hormone-releasing hormone (LHRH) fusion protein vaccine or surgical castration, at two years of age, on growth and carcass characteristics of Bos indicus bulls was evaluated. Seventy Nelore-cross bulls were divided into three groups: (1) immunized, (2) castrated and (3) intact control. At slaughter (three years of age), intact bulls had higher body weights, ADG, carcass weights, and muscle percentage compared to immunized and surgically castrated animals. Both castrated and immunized animals had greater marbling and percent carcass fat than the intact bulls. Average tenderness scores were inferior for intact bulls compared to immunized and castrated animals, but these differences were not significant (P>0.05). Juiciness, flavor, thawing, nor cooking losses differed significantly among the three groups. Immunocastration was effective in producing carcass traits similar to that of surgical castration. Therefore, immunization with LHRH fusion proteins appears to have practical utility in the management and castration of grazing bulls.

Relationships of gonadotropins, testosterone, and cortisol in response to GnRH and GnRH antagonist in boars selected for high and low follicle-stimulating hormone levels.

Considerable variation exists in the serum levels of gonadotropins in boars; this results in differential testicular function. Boars (Chinese Meishan, European White composite, and crosses of the two breeds) selected for high and low circulating FSH concentrations were used to define possible differences in pituitary sensitivity to GnRH and GnRH antagonist and gonadal and adrenal responses. After a 2-h pretreatment sampling period, boars were injected with GnRH or GnRH antagonist and repetitively sampled via jugular cannula for changes in serum concentrations of FSH, LH, testosterone, and cortisol. In response to varying doses of GnRH or GnRH antagonist, FSH, LH, or testosterone changes were not different in high- or low-FSH boars. Declines in LH after GnRH stimulation were consistently faster in boars selected for high FSH. Chinese Meishan boars had considerably higher cortisol concentrations than White composite boars (132.2 +/- 28.5 vs 67.4 +/- 26.8 ng/mL, respectively; P < .01). When select high- and low-gonadotropin Meishan:White composite crossbreds were sampled, cortisol levels were elevated but comparable between the two groups (126.5 +/- 13.7 vs 131.4 +/- 13.4 ng/mL, respectively). After GnRH antagonist lowered LH concentrations, administration of hCG resulted in increased testosterone and cortisol concentrations. Although testosterone concentrations remained high for 30 h, cortisol concentrations returned to normal levels within 10 h after hCG injection. The mechanism by which boars selected for high gonadotropins achieve increased levels of LH and FSH may not be due to differences in pituitary sensitivity to GnRH but to differences in clearance from the circulation.

GnRH antagonist inhibition of gonadotropin and steroid secretion in boars in vivo and steroid production in vitro.

The hormone GnRH has a stimulatory effect on gonadotropin synthesis and secretion. The objective of the first study was to evaluate concentrations of FSH and LH in plasma of boars after successive treatment with SB75, a GnRH antagonist. Thirteen boars greater than 1 yr of age (eight White Composite [WC] and five Meishan [MS]) were injected once daily with SB75 (10 microg/kg of body weight) for 4 d. Plasma concentrations of LH and testosterone (T) decreased after 1 h from the first dose of SB75. After 12 h of treatment, LH gradually returned to pretreatment concentrations, but T remained suppressed (< 2 ng/mL) until after the last injection of SB75. There was a modest, but significant, reduction in FSH during treatment with SB75. The prolonged inhibitory effect of SB75 on suppression of plasma T concentrations, in the presence of pretreatment concentrations of LH, implied direct effects of SB75 at the testis. In the second experiment, testicular tissue from adult boars was incubated in the presence of three doses of human chorionic gonadotropin (hCG; 0, .5, and 5 IU) with SB75 (250 ng/mL) or with Deslorelin, a GnRH agonist (500 ng/mL). Samples of media were collected every hour for 3 h, and concentrations of T and estrone (E1) were determined by RIA. Concentrations of T and E1 increased with time in response to treatment with hCG. Co-treatment with SB75 decreased media concentrations of T (P < .01) and E1 (P < .03) compared to controls (77.9 vs 85.7 +/- 2.0 and 4.7 vs 5.3 +/- .2 ng/g). In contrast, treatment with Deslorelin had no effect on the amount of T (P > .50) or E1 (P > .26) released with all dosages of hCG. These results indicate that a GnRH antagonist has a direct effect on the testis, decreasing amounts of T and E1 released from the Leydig cells; however, treatment with a GnRH agonist had no direct effect on release of these gonadal steroids. Thus, it remains unresolved whether the site of action of GnRH antagonist on testicular steroidogenesis is through a testicular GnRH receptor or through some other mechanism.

Estrus synchronization of beef cattle with a combination of melengestrol acetate and an injection of progesterone and 17beta-estradiol.

Our hypothesis was that estrus synchronization in beef cattle using melengestrol acetate (MGA) and an injection of progesterone (P4) and 17beta-estradiol (E2) to regress dominant ovarian follicles would improve pregnancy rate (number conceived/number in group) to AI compared with feeding only MGA or injecting PGF2alpha. During 2 yr, peripubertal heifers (n = 52) and cows (n = 327) received either 1) MGA for 18 d (d 0 = 1st d of MGA) plus an injection of P4 and E2 in sesame oil (vehicle) on d 11 to regress persistent ovarian follicles (MGA+P4), 2) MGA for 18 d plus vehicle on d 11 (MGA), or 3) two injections of PGF2alpha 10 d apart (d 7 and 17, PG). Concentration of P4 was assessed in blood samples obtained on d 0, 7, and 17 to indicate estrual status (anestrual or estrual) during treatment to induce estrus synchrony. Observations for detection of estrus occurred every 6 h for 180 h following treatment cessation. Females showing estrus were inseminated 6 to 12 h after estrus detection. Conception to AI was determined by ultrasonography 35 to 40 d later. Conception rate was greater (P < .05) in females in the PG than in those in the MGA group but did not differ from conception rate of females in the MGA+P4 group. Among anestrual females, estrus synchrony rates were greatest (P < .10) among females treated with MGA+P4. Among females that were estrual before treatment cessation, estrus synchrony rates were greater (P < .10) among females treated with MGA+P4 or PG than among those given MGA. Pregnancy rates were greater (P < .05) among females that were anestrual before treatment cessation and treated with MGA or MGA+P4 than among those treated with PG. Estrus synchronization using MGA+P4 and E2 differentially improves estrus synchronization and pregnancy rates among anestrual and estrual beef cattle while maintaining conception rates similar to those of PGF2alpha-treated females.

Pituitary relationship of gonadotropins and messenger ribonucleic acid for gonadotropin subunits in white composite and Meishan boars.

Two experiments were conducted to compare amounts of mRNAs for gonadotropin subunits in two breeds of pigs that are genetically distinct with respect to litter size: the Chinese Meishan and the white composite (a composite of four European white breeds: Yorkshire, Landrace, large white, and Chester white). In experiment 1, pituitaries were collected from mature Meishan (n = 5) and white composite (n = 6) boars. In experiment 2, boars were assigned to three groups according to peripheral concentrations of FSH: white composite (approximately 100 ng/ml; n = 8), Meishan with low FSH (< 500 ng/ml; n = 7), and Meishan with high FSH (> 750 ng/ml; n = 7). Within each of the three groups, five males were castrated. Two months after castration, pituitaries were collected from castrates and from eight contemporary boars for comparison purposes. Concentrations of FSH and LH were measured in half of each pituitary by RIA. RNA was isolated from the other half of each pituitary, applied to Northern and slot blots, and hybridized with cDNA probes for bovine alpha, LH beta, or FSH beta subunits. In experiment 1, pituitary concentrations of FSH (micrograms FSH/mg pituitary) were greater (p < 0.05) in Meishan (40 +/- 7 micrograms/mg; mean +/- SE) than in white composite boars (19 +/- 4). Pituitary weight and content of FSH were greater (p < 0.01) in Meishan boars (335.1 +/- 18 mg and 13.5 +/- 18 mg/pituitary, respectively) than in white composite boars (270.9 +/- 12.0 and 5.0 +/- 1.2, respectively). Northern analysis indicated that sizes of gonadotropin subunit mRNAs were similar in pigs and cattle. Amounts of mRNA for alpha subunit and LH beta subunit were not different (p = 0.14 and 0.08, respectively) between breeds. Meishans, however, had greater (p < 0.01) amounts of mRNA for FSH beta subunit than white composites. In experiment 2, amounts of mRNA for alpha, LH beta, and FSH beta subunits were not different (p > 0.05) between castrated and intact animals. Meishan males with high FSH had greater (p < 0.05) amounts of mRNA for all three gonadotropin subunits than did white composite males. From these results we suggest that greater levels of gonadotropins in the circulation of Meishan boars result, in part, from increased amounts of mRNA for gonadotropin subunits as compared with those in the white composite breed of boars.

The proximal 350 bp of 5'-flanking sequence of the human α-subunit glycoprotein hormone gene functions in the pituitary gland, but not the placenta, in transgenic mice.

To understand better the minimal DNA sequence requirements for regulated expression of the human α-subunit glycoprotein hormone gene (Hα), two lines of transgenic mice were constructed that contained a fusion gene (Hα-350CAT) consisting of only 350 bp of 5'-flanking sequence of Hα linked to the bacterial gene encoding chloramphenicol acetyltransferase (CAT). CAT activity was detectable in pituitary, but not in brain, heart, kidney, liver, lung, pancreas, or spleen in transgenic mice. Gonadectomy increased (p<0.05) CAT activity in the pituitaries of males (6.5±1.4% conversion/µg protein; mean ± SEM) and females (14.5±4.2) compared to intact males (1.2±0.3) and females (6.7±1.0). In addition, administration of a GnRH antagonist (antide; 60 µg/injection; one injection every other day) for 10 d to gonadectomized animals decreased (p<0.05) CAT activity in males (3.5±0.8) and females (2.9±0.5) compared to gonadectomized animals that received saline. Antide also reduced (p<0.05) serum concentrations of luteinizing hormone in gonadectomized males and females compared to gonadectomized animals that received saline. Surprisingly, CAT activity in the placenta of Hα-350CAT transgenic mice was not detectable (<3 SD above the mean of CAT activity in placenta from nontransgenic mice;n=77). Thus, expression of the human α-subunit promoter in the placenta of transgenic mice appears to require DNA sequences upstream of the proximal 350 bp of 5'-flanking sequence, whereas the proximal 350 bp of the human α-subunit gene contains sufficient DNA sequence to target pituitary-specific expression and confer responsiveness to gonadal hormones and GnRH.

Efficacy of a deletion mutant bovine herpesvirus-1 (BHV-1) vaccine that allows serologic differentiation of vaccinated from naturally infected animals.

Fifteen bovine herpesvirus-1 (BHV-1)-negative calves were vaccinated intramuscularly with 10(7.4) plaque-forming units of a double-deletion BHV-1 mutant (IBRV(NG)dltkdlgIII), and 6 remained as nonvaccinated controls. Thirty days after vaccination, the animals were challenged by nasal instillation of 10(8.2) CCID50 of a virulent BHV-1 strain (Cooper). The vaccinated calves were protected against wildtype virus challenge as demonstrated by clinical evaluation. Most of the vaccinates developed only a mild rhinitis (lasting an average of 6.5 days) with almost no systemic symptoms, whereas the controls developed a serious illness characterized by rhinitis (mean = 11.5 days), conjunctivitis, hyperthermia, apathy, loss of appetite, and dyspnea. The vaccinates also shed significantly less virus and for a shorter period of time (mean = 5.5 days) than the controls (mean = 9 days). Thirty days after vaccination, the vaccinates were negative in an anti-gIII specific blocking enzyme-linked immunosorbent assay (ELISA), despite the fact that most of them had developed neutralizing antibodies (serum neutralization titers ranging from 1:2 to 1:16). Seroconversion to gIII was detected as early as 7 days postinfection (dpi). Fourteen days after the challenge, all the animals exposed to wildtype BHV-1 had developed anti-gIII antibodies and were positive in this differential serologic test. Six controls plus 8 vaccinates kept in isolation were still positive to gIII when tested at 75 dpi. The use of the IBRV(NG)dltkdlgIII strain in conjunction with an anti-gIII specific blocking ELISA kit represents a powerful tool for BHV-1 control/eradication programs.