Molecular characterization of canine coronavirus strains circulating in Brazil

To characterize canine coronavirus (CCoV) circulating in diarrheic puppies in Brazil, 250 fecal samples collected between 2006 and 2012 were tested. By using RT-PCR to partially amplify the M gene, CCoV RNA was detected in 30 samples. Sequence analysis of the M protein grouped eight strains with CCoV-I and another 19 with CCoV-II prototypes. To genotype/subtype the CCoV strains and assess the occurrence of single or multiple CCoV infections, RT-PCR of the S gene was performed, and 25/30 CCoV-positive strains amplified with one or two primer pairs. For 17/25 samples, single infections were detected as follows: six CCoV-I, nine CCoV-IIa and two CCoV-IIb. Eight samples were positive for more than one genotype/subtype as follows: seven CCoV-I/IIa and one CCoV-I/IIb. Sequence analysis revealed that the CCoV-I and IIa strains shared high genetic similarity to each other and to the prototypes. The Brazilian strains of CCoV-IIb displayed an aminoacid insertion that was also described in CCoV-IIb-UCD-1 and TGEV strains. Among the 25 CCoV-positive puppies, five had a fatal outcome, all but one of which were cases of mixed infection. The current study is the first reported molecular characterization of CCoV-I, IIa and IIb strains in Brazil.     

Epidemiological Analysis of Leptospira spp. Infection in Equids from the Brejo Paraibano Microregion of Brazil

Leptospirosis is a zoonotic disease that occurs in humans and several animal species, including cattle, pigs, and horses, and it has a worldwide distribution. In equids, leptospirosis manifests through recurrent uveitis and abortion and other reproductive disorders. The objective of the present study was to investigate the epidemiological situation of Leptospira spp. infection in equids from the Brejo Paraibano microregion in the northeast of Brazil, that is, to assess the prevalence of and identify the major risk factors for infection. For this purpose, blood samples were taken from 257 equids (204 horses, 46 mules, and 7 donkeys), and a microscopic agglutination test (MAT) was used as serological analysis, with 24 serovars of Leptospira spp. as antigens. Antibodies were observed in 16.2% of horses, 13.0% of mules, and 28.6% of donkeys. Animals between the ages of 2.5 and 11 years had a risk of infection 0.4 times lower than those younger than 2.5 years old (odds ratio, 0.4; 95% confidence interval, 0.18-0.90). Further studies are needed to identify potential wild reservoirs and the main routes of infections.     

Digestion and metabolism of low and high residual feed intake Nellore bulls

Understanding the reasons why animals of similar performances have different feed requirements is important to increase profits for cattle producers and to decrease the environmental footprint of beef cattle production. This study was carried out aiming to identify the associations between residual feed intake (RFI) and animal performance, nutrient digestibility, and blood metabolites related to energy balance of young Nellore bulls during the finishing period. Animals previously classified as low (n?=?13) and high RFI (n?=?12), with average initial body weight of 398 kg and age of 503 days were used. Cattle were fed a high energy diet and were slaughtered when rib fat thickness measured by ultrasound between the 12th and 13th ribs reached the minimum of 4 mm. A completely randomized design was adopted, being data analyzed with a mixed model that included the random effect of slaughter group, the fixed effect of RFI class, and linear effect of the covariate feedlot time. No differences were found (p?>?0.10) between RFI classes for performance, dry matter, and nutrients intake. However, dry (p?=?0.0911) and organic matter (p?=?0.0876) digestibility tended to be lower, and digestibility of neutral detergent fiber corrected for ash and protein (p?=?0.0017), and total digestible nutrients (p?=?0.0657) were lower for high RFI animals, indicating lesser capacity of food utilization. Difference between low and high RFI animals was also found for blood cortisol at the end of the trial (p?=?0.0044), having low RFI animals lower cortisol concentrations. Differences in the ability to digest food can affect the efficiency of transforming feed into meat by Nellore cattle.     

Influence of different supplements and sugarcane (Saccharum officinarum L.) cultivars on intake, digestible variables and methane production of dairy heifers under tropical conditions

The sulphur hexafluoride (SF(6)) gas tracer method was used to measure methane (CH(4)) production of crossbred (3/4 Holstein x Zebu) dairy heifers fed two types of sugarcane (Saccharum officinarum L.; cultivar IAC-862480 (CC1) or cultivar IAC-873184 (CC2)) and supplemented with urea or concentrate. The study was performed at Embrapa Southeast Cattle, S?o Carlos, SP, Brazil, using a completely randomised design. Differences between treatments were significant for digestibility of dry matter, organic matter and energy. When animals were supplemented with urea differences between sugarcane cultivars did occur for NDF consumption, but not for daily methane production. This suggest that variation in chemical composition of sugarcane did not affect bovine ruminal CH(4) emissions. Concentrate inclusion in animal diet increased digestible organic matter intake, improving the nutrient intake by animals, but did not reduce CH(4) production expressed as a percentage of gross energy intake.     

Organic additives used in beef cattle feedlot: Effects on metabolic parameters and animal performance

Organic additives are recently being used in animal diets owing to their ability to control metabolic issues and result in better animal performance. Specifically, the organic additive Fator P^® presents an additional advantage that is to cause a lesser greenhouse gas emission. This study evaluated whether Fator P^® intake changes ruminal parameters or animal performance of beef cattle. Evaluations were carried out in a feedlot experiment divided into growing (46 days; two diets [control mix—CM and standard mix—SM] and finishing (lasted 83 days; four diets: CM, SM, Fator P^® + virginiamycin, and Fator P^® alone [FP]) trials. Animal performance study involved 48 animals allocated to 12 collective pens in completely randomized experimental design. Ruminal parameters were evaluated in separate metabolism study developed carried out using individual pen with four steers. During growing trial, FP diet resulted in higher (p < 0.05) dry matter intake (DMI) and ruminating time. In the finishing trial, diets containing Fator P^® resulted in higher DMI than obtained with CM. Most of the ruminal parameters did not differ (p > 0.05) among dietary treatments. Therefore, Fator P^® represents a viable and safe strategy for supplementation to beef cattle finished using high‐concentrate diet in feedlot systems.     

Trans-10, cis-12 conjugated linoleic acid and the PPAR-γ agonist rosiglitazone attenuate lipopolysaccharide-induced TNF-α production by bovine immune cells

Lipopolysaccharide (LPS) modulates innate immunity through alteration of cytokine production by immune cells. The objective of this study was to examine the effect of exogenous conjugated linoleic acid (CLA) and PPAR-γ agonist, rosiglitazone, on LPS-induced tumor necrosis factor α (TNF-α) production by cultured whole blood from prepubertal Holstein heifers (mean age, 5.5 mo). Compared with unstimulated cells, addition of LPS (10 μg/mL) to the culture medium increased (P < 0.03) peripheral blood mononuclear cell proliferation ≤2.5-fold. Coincubation with interferon γ (5 ng/mL) further stimulated (P < 0.01) the lymphoproliferative response to LPS. Lipopolysaccharide increased (P < 0.01) TNF-α concentration in cultured whole blood in a dose- and time-dependent manner. The greatest TNF-α stimulation occurred after 12 h of exposure to 1 μg/mL LPS. Coincubation with trans-10, cis-12 CLA isomer (100 μM) or rosiglitazone (10 μM), a PPAR-γ agonist, decreased (P < 0.01) LPS-induced TNF-α production by 13% and 29%, respectively. Linoleic acid and cis-9, trans-11 CLA isomer had no detectable effects on LPS-induced TNF-α production in cultured bovine blood. The PPAR-γ agonist-induced TNF-α attenuation was reversed when blood was treated with both rosiglitazone and GW9662, a selective PPAR-γ antagonist. Addition of rosiglitazone to the culture medium tended to reduce nuclear factor-κ Bp65 concentration in nuclear and cytosolic extracts isolated from cultured peripheral blood mononuclear cells. Results show that LPS is a potent inducer of TNF-α production in bovine blood cells and that trans-10, cis-12 CLA and PPAR-γ agonists may attenuate the pro-inflammatory response induced by LPS in growing dairy heifers. Additional studies are needed to fully characterize the involvement of nuclear factor-κ B in LPS signaling in bovine blood cells.     

Identification of the tet(B) resistance gene in Streptococcus suis

The tetracycline resistance gene, tet(B), has been described previously in Gram negative bacteria. In this study tet(B) was detected in plasmid extracts from 17/111 (15%) Streptococcus suis isolates from diseased pigs, representing the first report of this resistance gene in Gram positive bacteria.     

Effect of Chemical or Electrical Activation of Bovine Oocytes on Blastocyst Development and Quality

Activation of in vitro‐matured (IVM) oocytes is essential for successful embryo production following nuclear transfer (NT) or intracytoplasmic sperm injection (ICSI). This study was designed to compare the rates of blastocyst production and embryo quality (as measured by numbers of viable cells) following parthenogenetic activation with electrical pulse or the use of two different calcium ionophores, A23187 (CA) or ionomycin (IO), with or without the addition of bovine serum albumin (BSA). IVM oocytes with a first polar body were randomly allocated to five treatment groups: CA (5 μm CA, 5 min; n = 88), CA + BSA (5 μm CA, 5 min; BSA, 5 min; n = 90), IO (5 μm IO, 5 min; n = 91), IO + BSA (5 μm IO, 5 min; BSA, 5 min; n = 86) and EL (two pulses of 1.5 kV/cm, 20 μs; n = 120). Blastocyst rates were higher (p < 0.05) for CA (54.4%), IO (51.4%) and EL (54.5%) than for IO + BSA (18.3%). Treatment CA + BSA (39.8%) did not differ from the others. There was no difference (p > 0.05) among treatments in total number of cells. However, the percentage of viable cells was reduced in CA (49.9%), CA + BSA (45.8%), IO (64.9%), IO + BSA (50.9%) compared with EL (82.7%). In summary, the addition of BSA to the IO treatment had an adverse effect on blastocyst production rates. Although there was no difference between electrical stimulation and chemical activation on blastocyst production rates, electrical activation resulted in blastocysts with a higher percentage of viable cells.     

Bovine Oocyte Quality in Relation to Ultrastructural Characteristics of Zona Pellucida,Polyspermic Penetration and Developmental Competence

In the present study, the effect of bovine oocyte quality related to ultrastructural characteristics of zona pellucida (ZP), polyspermic penetration and embryo developmental competence was evaluated. Cumulus–oocyte complexes were punctioned from 453 ovaries, classified as 1, 2, 3 and 4 according to their morphological aspect, matured for 24 h and then divided into two groups. In group A, oocytes were fixed in 2.5% glutaraldehyde and 0.1 m sodium cacodylate and examined under a scanning electron microscope. Photomicrographs were taken and ZP’s pores were evaluated in squares of 6.4‐μm width. In group B, oocytes were fertilized in vitro. After 48 h, non‐cleaved oocytes were fixed for polyspermy evaluation. On days 7, 9 and 10, embryos were classified as developed (blastocysts and hatched blastocysts). Results showed that quality 1 oocytes revealed a ZP pore diameter of 0.50 ± 0.07 μm, which was smaller than the observed on oocytes of quality 2 (0.83 ± 0.10 μm), quality 3 (1.02 ± 0.22 μm) and quality 4 (1.38 ± 0.59 μm) (p ≤ 0.05). For In Vitro Fertilization (IVF), results showed that embryos originating from oocytes classed as 3 and 4 had lower cleavage rate (68.4% and 43.8%) than those belonging to class 1 and 2 (79.5% and 69.3%) (p ≤ 0.05). None oocyte classified as 3 and 4 developed to hatch blastocysts, while for oocytes belonging to quality 1 and 2, these values were, respectively, 15.2% and 12.5%. Concerning polyspermy, oocytes class 1 and 2 had lower polyspermic penetration than those belonging to class 3 and 4 (respectively 4.1%, 4.5%, 11.1% and 9.8%, for class 1, 2, 3 and 4). In conclusion, the present study demonstrated that oocytes with low qualities result in lower developmental competence and with high percentage of polyspermy after IVF, which can be the result of the ZP structure such as the number and the pore’s diameter.     

Evaluation of miltefosine for the treatment of dogs naturally infected with L. infantum (=L. chagasi) in Brazil

Dogs naturally infected with Leishmania Infantum (=L. chagasi) were treated with miltefosine using different therapeutic regimens. The animals were evaluated for clinical evolution, biochemical parameters, parasite load (by real-time PCR), cytokine levels and humoral response. After treatment and during the following 24 months, there was progressive clinical improvement and complete recovery in 50% (7/14) of the treated animals. There was a decrease in the smear positivity of the bone marrow after treatment, and there was also a gradual and constant decrease in positive cultures at the end of the follow-up period. However, the PCR detection of parasite DNA remained positive. In general, all animals presented a significant increase in parasite load 6 months after treatment. The IFN-γ levels in all the groups tended to increase during follow-up period, regardless of the miltefosine dose administered. The IL-4 and IL-10 levels of the animals tended to decrease during follow-up, except after 300 days when only IL-10 increased. The serum antibodies identified antigens that ranged from 116 kDa to less than 29 kDa in the Western blot assay. Furthermore, 300 days after treatment, qualitative and quantitative differences in the antigen profiles were observed. Antigens of 97 and 46 kDa were the most intensely recognized. Higher levels of antigen-specific Leishmania IgG were detected before and 300 days after treatment in all groups. Taking together, the improvement in the clinical symptoms was not followed by parasitological clearance, suggesting that treatment with miltefosine is not recommended, especially in endemic areas like Brazil, where children are the major victims and dogs are involved in the maintenance of the parasite cycle.