Simultaneous detection and genotyping of porcine reproductive and respiratory syndrome virus (PRRSV) by real-time RT-PCR and amplicon melting curve analysis using SYBR Green

The feasibility of using a SYBR Green-based real-time RT-PCR assay (SYBR Green ReTi RT-PCR) followed by melting curve analysis (MCA) for detecting and genotyping porcine reproductive and respiratory syndrome virus (PRRSV) was assessed. The SYBR Green ReTi RT-PCR and a previously reported two-step, non-nested RT-PCR assays were simultaneously tested on selected European (EU) and North American (US) PRRSV strains and isolates collected from diverse clinical, temporal, and geographical origins. The validation experiments showed that the optimised SYBR Green ReTi RT-PCR can sensitively and specifically detect PRRSV, consistently detecting as little as 0.03TCID(50)/sample of each virus genotype, with no type-bias and no amplification signal for other swine pathogens. After MCA, two well-differentiated melting temperature (T(m)) profiles for each virus genotype were obtained, as sequencing confirmed it. High repeatability was obtained for the T(m) values, with intra-run coefficients of variation (CoVs) of 0.25 and 0.32 and inter-run CoVs of 0.42 and 0.52 for EU and US genotypes, respectively. The sensitivity of the SYBR Green ReTi RT-PCR (100%) was higher than that of the RT-PCR (95.7%) when testing field isolates. This greater sensitivity of the SYBR Green ReTi RT-PCR was further confirmed by the detection of a higher proportion of PRRSV-positive diagnostic specimens (29.7%) than by the RT-PCR (28.5%). The SYBR Green ReTi RT-PCR test detected infection as early as 2 dpi in the sera of experimentally infected pigs regardless of virus genotype, and discriminated negative (non-inoculated), EU- and US-infected pigs. In conclusion, the reported SYBR Green ReTi RT-PCR assay coupled with MCA can detect and type PRRSV and may be useful as an alternative diagnostic assay in diverse PRRSV epidemiological circumstances.     

Genetic associations between accumulated productivity, and reproductive and growth traits in Nelore cattle

The total meat yield in a beef cattle production cycle is economically very important and depends on the number of calves born per year or birth season, being directly related to reproductive potential. Accumulated Productivity (ACP) is an index that expresses a cow's capacity to give birth regularly at a young age and to wean animals of greater body weight. Using data from cattle participating in the “Program for Genetic Improvement of the Nelore Breed” (PMGRN — Nelore Brasil), bi-trait analyses were performed using the Restricted Maximum Likelihood method based on an ACP animal model and the following traits: age at first calving (AFC), female body weight adjusted for 365 (BW365) and 450 (BW450) days of age, and male scrotal circumference adjusted for 365 (SC365), 450 (SC450), 550 (SC550) and 730 (SC730) days of age. Median estimated ACP heritability was 0.19 and the genetic correlations with AFC, BW365, BW450, SC365, SC450, SC550 and SC730 were 0.33, 0.70, 0.65, 0.08, 0.07, 0.12 and 0.16, respectively. ACP increased and AFC decreased over time, revealing that the selection criteria genetically improved these traits. Selection based on ACP appears to favor the heaviest females at 365 and 450 days of age who showed better reproductive performance as regards AFC. Scrotal circumference was not genetically associated with ACP.     

Effects of six carbohydrate sources on diet digestibility and postprandial glucose and insulin responses in cats

The effects of diets with different starch sources on the total tract apparent digestibility and glucose and insulin responses in cats were investigated. Six experimental diets consisting of 35% starch were extruded, each containing one of the following ingredients: cassava flour, brewers rice, corn, sorghum, peas, or lentils. The experiment was carried out on 36 cats with 6 replications per diet in a completely randomized block design. The brewers rice diet offered greater DM, OM, and GE digestibility than the sorghum, corn, lentil, and pea diets (P < 0.05). For starch digestibility, the brewers rice diet had greater values (98.6%) than the sorghum (93.9%), lentil (95.2%), and pea (96.3%) diets (P < 0.05); however, starch digestibility was >93% for all the diets, proving that despite the low carbohydrate content of carnivorous diets, cats can efficiently digest this nutrient when it is properly processed into kibble. Mean and maximum glucose concentration and area under the glucose curve were greater for the corn-based diet than the cassava flour, sorghum, lentil, and pea diets (P < 0.05). The corn-based diets led to greater values for the mean glucose incremental concentration (10.2 mg/dL), maximum glucose incremental concentration (24.8 mg/dL), and area under the incremental glucose curve (185.5 mg.dL(-1).h(-1)) than the lentil diet (2.9 mg/dL, 3.1 mg/dL, and -40.4 mg.dL(-1).h(-1), respectively; P < 0.05). When compared with baseline values, only the corn diet stimulated an increase in the glucose response, occurring at 4 and 10 h postmeal (P < 0.05). The corn-based diet resulted in greater values for maximum incremental insulin concentration and area under the incremental insulin curve than the lentil-based diet (P < 0.05). However, plasma insulin concentrations rose in relation to the basal values for cats fed corn, sorghum, pea, and brewers rice diets (P < 0.05). Variations in diet digestibility and postprandial response can be explained by differences in the chemical composition of the starch source, including fiber content and granule structure, and also differences in the chemical compositions of the diets. The data suggest that starch has less of an effect on the cat postprandial glucose and insulin responses than on those of dogs and humans. This can be explained by the metabolic peculiarities of felines, which may slow and prolong starch digestion and absorption, leading to the delayed, less pronounced effects on their blood responses.     

Genital lesions and distribution of amastigotes in bitches naturally infected with Leishmania chagasi

Recent reports indicate that Leishmania chagasi has tropism to the male canine genital system, which is associated with shedding of the organism in the semen, supporting the hypothesis of venereal transmission. The aim of this study was to describe the lesions and assess parasite load in the genital system of bitches with canine visceral leishmaniasis (CanL). Symptomatic (n=5) and asymptomatic (n=5) bitches seropositive for CanL were randomly selected at the Center for Zoonosis Control (Belo Horizonte, State of Minas Gerais, Brazil). Five serologically negative, healthy, adult bitches also from the CZC were used as controls. Samples from genital organs (vulva, vagina, cervix, uterine body, uterine horns, uterine tubes, and ovaries), liver, and spleen were histologically evaluated and processed for immunodetection of Leishmania sp., and PCR. The most significant histological change was a mild to moderate vulvar dermatitis, characterized by a histio-plasma-lymphocytic infiltrate. This change was detected in all asymptomatic, four symptomatic, and three uninfected control bitches. In one symptomatic and one asymptomatic bitch intracytoplasmic amastigotes were observed within macrophages in the inflammatory infiltrate. Samples from all the segments of the genital tract were positive in at least one infected animal, in the absence of detectable amastigotes in the tissue. These findings support the notion that L. chagasi does not have genital tropism in the bitch, which is in contrast to our previous findings in naturally infected male intact dogs.     

Cross-sectional serological study of canine Leishmania infection in Fortaleza, Ceará state, Brazil

Visceral leishmaniasis (VL) is one of the most important reemerging parasitic disease in the world. The domestic dog is the main reservoir in urban environments. The aim of this work was to extend the knowledge on canine Leishmania infection in the city of Fortaleza in northeastern Brazil, identifying the risk factors inherent in dog susceptibility to the infection. Two populations were analyzed, domestic dogs from clinics and the Veterinary Hospital Unit of Ceará State University and stray dogs captured by the Center for Zoonosis Control in Fortaleza. Blood samples were collected and centrifuged and the sera were stored at -20 degrees C. ELISA, with soluble crude Leishmania chagasi antigens (LTCC - WDCM731) was used for diagnosis. A total of 1,381 samples were tested, 750 from domestic and 631 from stray dogs. The seroprevalence of canine VL was 21.4% (135/631) in stray dogs and 26.2% (197/750) in domestic dogs. The seroprevalence of Leishmania infection in the six administrative regions of the city (Secretarias Executivas Regionais, or SER) among stray dogs was highest in SER V, representing 31.4% of the cases, with large dogs more infected (27.7%). Among domestic dogs Leishmania infection was most prevalent in SER V (38.5%) and VI (37.6%). The dogs' age (1-6 years), large size, environment with dense vegetation and presence of clinical signs compatible with Leishmania infection were associated with the illness in domestic dogs. The frequency of the infection varied seasonally. The seroprevalence was greatest in July and December. These results confirm Fortaleza is an endemic area for canine VL and suggest some variables associated with increasing infection risk in dog populations.     

Acaricidal activity of limonene, limonene oxide and beta-amino alcohol derivatives on Rhipicephalus (Boophilus) microplus

Limonene, limonene oxide and eight beta-amino alcohol derivatives obtained by synthesis were investigated for the effect on egg hatchability and mortality rates of newly hatched larvae of the cattle tick Rhipicephalus (Boophilus) microplus. At the doses between 10mug/ml and 2.5mug/ml all the compounds were highly lethal to the larvae and some of them showed activity at lower concentrations. The effect on the eggs hatchability was observed in all the treatments.     

Surgical Management of Cecal Impaction/Dysfunction by Ileocolostomy in a Horse

A four-year-old Appaloosa brood mare was referred for evaluation of a colic episode of 24 hours' duration. Physical examination revealed a distended cecum filled predominantly with fluid. A tentative diagnosis of cecal dysfunction was determined, and surgical intervention was performed by means of typhlotomy and ileocolostomy techniques through a complete bypass of the cecum. Postanaesthetic myopathy and incisional infection developed as postoperative complications. The mare was discharged 13 days after surgery. Follow-up information obtained from the owner and referring veterinarian 4 months after discharge revealed full recovery of the mare.     

Supplementation of pig diets in the growth and termination phases with different calcium sources

The aim of this study was to evaluate the effect of supplementation of pig diets in the growth and termination phases with different calcium sources. In experiment I, 36 whole males were distributed in randomized blocks in six groups, with six replications. A basal diet was formulated to meet the animals’ nutritional requirements except for calcium (0.09%), and the sources evaluated (calcitic limestone, monodicalcium phosphate, calcinated bone flour, and oyster flour) replaced the basal diet to provide 0.59% of total calcium. To determine the endogenous calcium, a diet containing low calcium (0.019%) was given simultaneously to another group of animals. Feces and urine were collected for determination the coefficients of apparent and true digestibility. In experiment II, 160 piglets were distributed in randomized blocks in four treatments, with five replications and four animals per experimental unit. Carcass and performance parameters, calcium concentration in bone and serum, and bone parameters were evaluated. The data were submitted to analysis of variance and factorial. The calcium source did not influence the digestibility coefficients determined by total collection (P > 0.05). The digestibility of Ca from oyster flour estimated by collection with an indicator was higher than that from the other sources (P < 0.05). Calcium sources did not interfere in the evaluated parameters (P > 0.05). The sources studied in this work can be used to supplement growing pigs’ diets.     

Replacement of Tifton hay by spineless cactus in Girolando post-weaned heifers´ diets

This study aims to evaluate the effect of the Tifton 85 hay (Cynodon dactylon L. Pers) replacement by Orelha de Elefante Mexicana spineless cactus (Opuntia stricta [Haw.] Haw) on the nutrient intake and digestibility, growth performance, microbial protein synthesis, and efficiency of dietary nitrogen compound utilization in the diets of Girolando heifers. Twenty-four (3.5 months and 100 kg ± 3.5 kg) Girolando heifers (5/8 Holstein x Gyr) were used and arranged in a completely randomized design. Dry matter, organic matter, crude protein, and neutral detergent fiber decreased linearly as a function of the levels of Tifton hay (TH) replacement by Orelha de Elefante Mexicana spineless cactus (OEM). The non-fiber carbohydrate intake and coefficient of dry matter digestibility increased, while the neutral detergent fiber (NDF) digestibility decreased linearly with the OEM inclusion. After evaluating the growth performance by observing weight gain and morphometric measurements, there were no significant alterations. The urinary volume, nitrogen balance as well as the efficiency of nitrogen retention had increased, while the urinary and plasma urea concentrations had decreased linearly with the OEM inclusion. The results obtained in this study show that Orelha de Elefante Mexicana spineless cactus should replace Tifton hay in post-weaned heifer diets.     

Different roughage:concentrate ratios with and without liquid residue of cassava for lambs

This aim of this study was to evaluate the intake and apparent digestibility of nutrients, ingestive behavior, blood parameters, and performance of lambs fed two roughage:concentrate (R:C) ratios, with or without liquid residue of cassava (LRC). Forty lambs (19.5?±?1.45 kg body weight) were distributed in a completely randomized 2?×?2 factorial design. The intake of dry matter (1311 g/day) and total digestible nutrients (755 g/day), and the intake (172 g/day) and digestibility (552 g/kg) of crude protein were higher for lambs fed R:C of 40:60. The R:C ratio of 80:20 promoted a higher intake (585 g/day) and digestibility (461 g/kg) of neutral detergent fiber. There was an intake of LRC of 1.37 L/day for lambs fed R:C of 80:20 and 0.42 L/day for lambs fed R:C of 40:60. The lambs fed R:C of 80:20 spent more time in feeding (326 m/day) and rumination (530 m/day). The metabolic profile and performance of lambs were not affected by LRC inclusion. The body weight gain (220 g/day) was higher in lambs fed R:C of 40:60. A roughage:concentrate ratio of 40:60 is recommended for lambs because it resulted in the best performance, regardless of supplementation with liquid residue of cassava.