Validation of RT‐PCR Assays for Molecular Characterization of Porcine Teschoviruses and Enteroviruses

Porcine enteroviruses (PEVs) and teschoviruses (PTVs) are described as causative agents of neurological disorders, fertility disorders and dermal lesions of swine. Difficulties in the serological detection of these viruses may lead to a significant underestimation of infections with clinical symptoms. With the recent availability of genome sequence data for all the serotypes, molecular diagnosis is a possibility. The present study describes a new approach to molecular ‘serotyping’ of PTVs and PEV‐B viruses, involving the amplification and sequencing of a genomic fragment of the VP1 coding region. A molecular characterization of Italian entero‐teschovirus isolates was performed using a set of previously published and newly designed polymerase chain reaction primers. A total of 33 porcine isolates and 10 reference strains were analysed. Porcine enterovirus‐B samples were first diagnosed as positive for enterovirus by amplification of the 5′‐non‐translated region. Samples were then typed by amplification and sequencing of a portion of the VP1 coding region. Porcine enterovirus‐A and PTVs were detected by a published assay in the 5′‐NC region that allows them to be differentiated according to the size of amplification product, using the same set of primers. For serotype characterization of PTV, we evaluated four different regions: the N terminus of the capsid protein VP2, the region encoding for RNA‐dependent RNA polymerase, and the capsid VP1 and VP4 regions. The newly designed primers in the VP1 region was proved to be broad in range and suitable for serotype assessment and therefore constitute a useful diagnostic tool for molecular diagnosis of porcine teschovirus/enterovirus strains and for the study of molecular epidemiology and evolution of these viruses.     

Higher feeding diets effects on age and liveweight gain at puberty in crossbred Nelore × Hereford heifers

This study was conducted to determine the age and liveweight at puberty of 120 crossbred beef heifers submitted to four diets to achieve predetermined weight gains (kg/day): 0.5 (G500; n?=?32), 0.75 (G750; n?=?32), 1.0 (G1000; n?=?29), and 1.25 (G1250; n?=?27). Animals were classified depending on their level of crossing between Nelore (N) and Hereford (H): 25%N-75%H, 37.5%N-63.5%H, 43.7%N-56.7%H, 50%N-50%H, and 75%N-25%H. Reproductive evaluation was performed at the beginning and at the end of the experimental period by ultrasonography and hormone analysis. The average age and liveweight at puberty were 388.0?±?1.9 days and 331.4?±?1.3 kg, respectively. Animals from the 25%N-75%H group reached puberty earlier than heifers from other genetic groups supplemented with G1250 diet (P?<?0.05). Heifers with higher degree of Nelore (75%N-25%H) fed with G1000 diet showed estrus 42 days prior to mating, but only 57 % reached puberty at mating (P?<?0.05). Heifers with follicles of higher diameter reached puberty (P?<?0.05) earlier. Higher average daily weight gain showed a positive effect on follicular diameter and IGF-I level at puberty (P?<?0.01). Concentrations of GH were lower in heifers fed G1250 compared to G1000 diet (P?<?0.05). There was a significant interaction between nutritional level and insulin levels at puberty (P?<?0.01). We demonstrated the relationship between IGF-I and average daily gain on the onset of puberty in heifers. In conclusion, heifers submitted to the higher feeding level showed a higher follicular diameter and were younger at puberty.     

Adjusted method of penis fixation during boar semi-automatic semen collection aiming to reduce bacterial contamination

Semen collection has an essential role in the initial bacterial load in boar ejaculates and extended semen. The study aimed to explore the efficacy of an adjusted penis fixation in a semi-automatic collection system on reducing bacterial contamination of ejaculates in two-boar studs with different scenarios. Historically, stud A had low levels of bacterial load in raw semen, while stud B had a high level of contamination. A total of 56 mature boars had their semen collected using two methods of penis fixation: (a) Traditional: The penis was fixed directly with the artificial cervix and transferred to the adjustable clamp; (b) Adjusted: The fixation was performed with one gloved-hand, and after exteriorization, the penis was gripped using the artificial cervix with the other gloved-hand and transferred to the adjustable clamp. The bacterial load (p = .0045) and the occurrence of ejaculates >231 CFU/ml (p = .0101) were reduced in the Adjusted compared to the Traditional method. Bacterial load was reduced when using the Adjusted method in stud B (p = .0011), which showed a greater occurrence of critical factors for bacterial contamination (p ≤ .0034). The Adjusted method reduced the occurrence of ejaculates >231 CFU/ml when the preputial ostium was dirty (p = .016) and the duration of semen collection was >7 min (p = .022) compared to the Traditional method. In conclusion, the Adjusted penis fixation was efficient in reducing bacterial load of ejaculates, mainly in boar stud B, which had high contamination challenges.     

The effect of life history events on carcass merit and price of cull dairy cows

Cull dairy cows contribute almost 10% of national beef production in the United States. However, different factors throughout the life of dairy cows affect their weight and overall body condition as well as carcass traits, and consequently affect their market price. Therefore, the objectives of this study were: (1) to assess relationships between price ratio and carcass merit of cull dairy cows sold through several sites of an auction market and (2) to investigate the effect of animal life history events and live weight on sale barn price (BP) and price ratio (as a measure of relative price), as an indicator of carcass merit. Data from 4 dairy operations included 3,602 cull dairy cow records during the period of 2015 to 2019. Life history events data were collected from each dairy operation through Dairy Comp software; live weight and price were obtained periodically from the auction market, and the carcass data were provided by a local packing plant. Cow price in dollars per unit of live weight ($/cwt) and price ratio were the 2 outcome variables used in the analyses. Price ratio was created aiming to remove seasonality effects from BP (BP divided by the national average price for its respective month and year of sale). The association between price ratio and carcass merit traits was investigated using canonical correlation analysis, and the effect of life history events on both BP and price ratio was inferred using a multiple linear regression technique. More than 70% of the cows were culled in the first 3 lactations, with an average live weight of 701.5 kg, carcass weight of 325 kg, and dressing percentage of 46.3%. On average, cull cows were sold at $57.0/cwt during the period considered. The canonical correlation between price ratio and carcass merit traits was 0.76, indicating that price ratio reflected carcass merit of cull cows. Later lactations led to lower BP compared with cows culled during the first 2 lactations. Injury, and leg and feet problems negatively affected BP. Productive variables demonstrated that the greater milk production might lead to lower cow prices. A large variation between farms was also noted. In conclusion, price ratio was a good indicator of carcass merit of cull cows, and life history events significantly affected sale BP and carcass merit of cull cows sold through auction markets.     

Genetic variability following selection for scrapie resistance in six autochthonous sheep breeds in the province of Bolzano (northern Italy)

Scrapie is an ovine transmissible spongiform encephalopathy, and its susceptibility is associated with polymorphisms in the prion protein gene (PRNP). Genetic selection is currently the most effective mean for eradication of the susceptible VRQ allele in favour of resistant ARR allele. Maintenance of genetic diversity should be one of the major objectives in breeding programmes, especially in endangered breeds, and genetic information are an excellent alternative to pedigree data where these information are missing. The aim of our study was to determine changes of genetic variability in six native sheep breeds from autonomous province of Bolzano, northern Italy, following simulation of scrapie selection scenarios. A total of 684 rams were investigated for PRNP polymorphisms and for 10 microsatellite loci to estimate genetic variability. Across all loci, a total of 163 alleles were detected with a mean of 10.4 alleles per locus. Average observed (Ho) and unbiased expected (uHe) heterozygosity overall loci were 0.74 and 0.78, respectively, showing a statistically significant deviation from Hardy–Weinberg equilibrium (HWE) in all breeds. This heterozygosity deficit was confirmed by a positive fixation index (Fis), determining a moderate inbreeding in each breed. Simulating a soft selection, where only rams having at least a VRQ allele should be excluded from reproduction, Ho, uHe and Fis values remained almost unchanged, indicating that genetic variability should not be affected by the removal of these individuals. With a mild selection scenario, considering only rams with at least one ARR allele, we observed a decrease in the mean alleles per breed (8.9) and the maintenance of heterozygosity deficiency, except for two breeds, where it was any longer significant. These results showed that selection strategies allowing use of heterozygous as well homozygous ARR rams might be the right compromise to improve resistance to scrapie and to do not dramatically affect genetic variability of these breeds.     

Effects of protein-energetic supplementation frequency on growth performance and nutritional characteristics of grazing beef cattle

Two experiments were conducted to evaluate reduced supplementation frequencies for grazing beef cattle in rainy season. In experiment 1, evaluating the nutritional parameters, four rumen-cannulated Nellore bulls (BW = 410 kg) were used. In experiment 2, evaluating animal performance, 48 Nellore bulls (BW = 358 kg) were used. The treatments were as follows: mineral supplement (MS) alone and MS plus protein-energy supplement provided 3×, 5× and 7×/week. Supplementation frequency did not affect (P > 0.05) intake and digestibility. Average daily gain was greater (P < 0.001) to supplementation compared with MS. The supplementation 5×/week resulted in greater weight gain per hectare (9.24) and higher economic returns during the study period (1.64%) compared to other supplementations. Supplementation 5×/week increased animal performance and positively influenced economic returns.     

Bacterial pseudomycetoma in dwarf hamster, Phodopus sungorus

A case of a dwarf hamster with two progressively growing nodules on the right fore limb is described. These were excised following ineffective medical treatment and were submitted for histopathological examination, which revealed bacterial pseudomycetoma in both nodules. To the authors' knowledge this is the first reported case of bacterial pseudomycetoma in a dwarf hamster.     

Adenosine deaminase activity in serum, erythrocytes and lymphocytes of rats infected with Leptospira icterohaemorrhagiae

Leptospirosis is a systemic disease of humans and domestic animals, mainly dogs, cattle and swine. The course of human leptospirosis varies from mild to severe fatal forms and the most severe form of human leptospirosis is principally caused by Leptospira interrogans serovar icterohaemorrhagiae (L. icterohaemorrhagiae). The enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. The aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with L. icterohaemorrhagiae, as compared with non-infected rats. Twenty-four adult rats, divided into two uniform groups (A and B) were used for the enzymatic assays. The animals in Group B were inoculated intraperitoneally with 2×10(8) leptospires/rat, and the rodents in Group A (control) were not-inoculated. Blood collection was performed on days 5 and 15 post-infection (PI) and the blood used to assess the ADA activity. The infection by L.icterohaemorrhagiae altered erythrocyte count, hemoglobin concentration and hematocrit, causing a decrease in all these parameters on day 15 PI. Lymphocytes decreased significantly on day 15 PI, and ADA activity in serum was inhibited in infected rats on days 5 and 15 PI and its activity in erythrocytes were increased on day 5 PI. On day 5 PI, we found an increase in ADA activity in erythrocytes of infected rats. No correlation was observed between hematocrit and erythrocyte ADA activity on days 5 and 15 PI. The ADA activity was inhibited in rats infected on day 15 PI. A positive correlation (r(2)=60) was also observed between the number of lymphocytes and ADA activity in lymphocytes on day 15 PI (P<0.05). In conclusion, our results showed that the ADA activity is altered in serum, lymphocytes and erythrocytes in experimental infection by L.icterohaemorrhagiae in rats, concomitantly with hematological parameters.     

Multiple-breed genetic inference using heavy-tailed structural models for heterogeneous residual variances

Multiple-breed genetic models recently have been demonstrated to account for the heterogenous genetic variances that exist between different beef cattle breed groups. We extend these models to allow for residual heteroskedasticity (heterogeneous residual variances), specified as a function of fixed effects (e.g., sex, breed proportion, breed group heterozygosity) and random effects such as contemporary groups (CG). We additionally specify the residual distributions to be either Gaussian or based on heavier-tailed alternatives such as the Student's t or Slash densities. For each of these three residual densities using either homoskedastic (homogeneous variance) or heteroskedastic error specifications, we analyzed 22,717 postweaning gain records from a Nelore-Hereford population based on a Markov chain Monte Carlo animal model implementation. The heteroskedastic Student's t error model (with estimated df = 7.33 +/- 0.48) was clearly the best-fitting model based on a pseudo-Bayes factor criterion. Breed group heterozygosity and, to a lesser extent, calf sex seemed to be marginally important sources of residual heteroskedasticity. Specifically, the residual variance in F1 animals was estimated to be 0.70 +/- 0.16 times that for purebreds, whereas the male residual variance was estimated to be 1.13 +/- 0.09 times that for females. The CG effects were important random sources of residual heteroskedasticity (i.e., the coefficient of variation of CG-specific residual variances was estimated to be 0.72 +/- 0.06). Purebred Nelores were estimated to have a larger genetic variance (124.84 +/- 21.75 kg2) compared with Herefords (40.89 +/- 6.70 kg2) under the heteroskedastic Student's t error model; however, the converse was observed from results based on a homoskedastic Student's t error model (46.24 +/- 10.90 kg2 and 60.11 +/- 8.54 kg2, respectively). These results indicate that allowing for robustness to outliers and accounting for heteroskedasticity of residual variances has potentially important implications for variance component and genetic parameter estimates from data on multiple-breed populations.