Cloning and purification of the Streptococcus suis serotype 2 glyceraldehyde-3-phosphate dehydrogenase and its involvement as an adhesin

Streptococcus suis serotype 2 is a swine pathogen responsible for diverse diseases and may be present in the tonsils of pigs which show no sign of illness. Because adhesion to host cells may be important in the carrier state, this study was undertaken to characterize a 39 kDa surface protein identified as a glyceraldehyde-3-phosphate dehydrogenase (GAPDH), possibly implicated in the adhesion of the bacteria. The gene encoding for the GAPDH of S. suis was cloned and sequenced. The DNA sequence contained an open reading frame encoding for a 336 amino acid polypeptide exhibiting 95% sequence identity with the GAPDH from Streptococcus pyogenes and from other streptococci. Using the Qiaexpress expression plasmids, the gapdh gene was inducibly overexpressed in E. coli to produce GAPDH with a hexahistidyl N-terminus to permit its purification. The (His)6GAPDH protein was found to possess functional GAPDH enzymatic activity after the purification. An adherence assay with S. suis and porcine tracheal rings pre-incubated with (His)6GAPDH and non-incubated rings was showed a significant reduction in the adhesion of S. suis in the (His)6GAPDH pre-incubated rings compared to the non-incubated rings. The GAPDH protein of S. suis seems to be involved in the first steps of the bacterial adhesion to host cells.     

The ischiorectal fossa: an alternative route for the administration of prostaglandin in cattle

Three experiments were conducted to investigate the ischiorectal fossa (IRF) as a route for the administration of prostaglandin F2 alpha (dinoprost) in cattle. In Experiment 1, 21 nonlactating Holstein cows were given 100 micrograms of gonadotropin releasing hormone (GnRH), intramuscularly (i.m.), and, 7 d later, 25 mg of dinoprost into the IRF. Sixteen cows had serum progesterone concentrations > or = 1.0 ng/mL at the time of dinoprost treatment, and all of these had rapid and complete luteolysis; the other 5 cows were not considered to have a functional corpus luteum (CL) at the time of treatment. There were minimal adverse behavioral reactions to the IRF injections and no visible or palpable tissue reactions at the injection site. In Experiment 2, 74 Holstein heifers were given 25 mg of dinoprost by IRF injection. Luteolysis occurred in 84.3% of the heifers with a functional CL (as determined by the serum progesterone concentration at the time of treatment). Of the heifers bred by either natural service or artificial insemination, 61.8% became pregnant. In Experiment 3, 48 beef heifers received dinoprost 7 d after ovulation, as follows: 25 mg, i.m. (n = 9); 25 mg, IRF (n = 10); 10 mg, IRF (n = 10); 10 mg, subcutaneously (s.c.) (n = 10); or 10 mg, intravulvosubmucosally (IVSM) (n = 9). Fewer heifers (P < 0.05) were found to be in estrus or ovulating in the 10 mg IVSM group (0% and 11%, respectively) than in the 25 mg i.m. group (100% and 100%), the 25 mg IRF group (90% and 100%, respectively), or the 10 mg IRF group (80% and 80%). The rates of estrus (50%) and ovulation (50%) were intermediate in the 10 mg s.c. group. In summary, 25 mg of dinoprost injected into the IRF caused minimal behavioral or tissue response and induced luteolysis and fertile estrus. In addition, 10 mg of dinoprost injected into the IRF was as efficacious as 25 mg given either i.m. or into the IRF in inducing estrus and ovulation.     

Clonal relationships among avian Escherichia coli isolates determined by enterobacterial repetitive intergenic consensus (ERIC)-PCR

Forty-nine avian Escherichia coli isolates isolated from different outbreak cases of septicemia (24 isolates), swollen head syndrome (14 isolates) and omphalitis (11 isolates), and 30 commensal isolates isolated from poultry with no signs of illness were characterized by enterobacterial repetitive intergenic consensus (ERIC)-PCR technique and their serotypes were determined. The ERIC-PCR profile allowed the typing of the 79 isolates into 68 ERIC-types and grouped the isolates into four main clusters (A-D), with the omphalitis isolates being grouped with the commensals and separated from the septicaemia and swollen head syndrome. These results indicate that ERIC-PCR is a technique that could replace other molecular characterization techniques such as random amplification of polymorphic DNA (RAPD)-PCR and restriction fragment length polymorphism (RFLP), reinforce previous observations that omphalitis isolates are just opportunistic agents, and are consistent with many reports that specific genotypes are responsible for causing specific diseases. Most of the isolates were either nontypable or rough, supporting the need for alternative methods for typing these isolates.     

Clinical and hematological signs associated with dogs naturally infected by Hepatozoon sp. and with other hematozoa: a retrospective study in Uberlândia, Minas Gerais, Brazil

An evaluation was made of the clinical and hematological aspects of 115 dogs infected naturally by Hepatozoon sp. and treated at the Hospital School of the Faculty of Veterinary Medicine in Uberlandia, MG, Brazil. Of the 115 dogs for whom peripheral blood films were evaluated, 89 (77.39%) presented parasitemia by Hepatozoon sp. solely, while 26 (22.61%) had combination of Hepatozoon sp., Babesia sp. and Ehrlichia sp. Young male dogs less than a year old, of undefined breed (UB), were the most commonly affected. Thirty-nine (33.92%) of the dogs were asymptomatic while 76 (66.08%) presented varied clinical symptoms, the most frequent being anorexia, pulmonary alterations, hyperthermia, pale mucosae, apathy and/or prostration, and diarrhea. The majority of hematological alterations were normochromic-normocytic anemia, leukocytosis, neutrophilia, and nuclear deviation of neutrophils to the left (NDNL). The findings of this study confirm that Hepatozoon sp. causes clinical and hematological alterations of varied intensity, which, albeit not specific to canine hepatozoonosis, reinforce the notion that the discovery of the agent in dogs, even with low parasitemia, should be taken into consideration.     

Energy balance in grazing Jersey cows in early lactation supplemented with peanut and sunflower oils

In this study, we evaluated the effects of supplementation with peanut and sunflower oils on intake and digestibility, milk yield and composition, energy balance (EB), changes in weight and body condition score (BW and BCS), and blood metabolites of Jersey cows on pasture in early lactation. Twenty-four cows were distributed in a randomized block design where they received the following treatments: concentrate without oil (CON), concentrate plus 59.6 g/kg DM peanut oil (PEA), concentrate plus 59.6 g/kg DM sunflower oil (SUN), and concentrate plus 59.6 g/kg DM of a 1:1 mixture of peanut oil and sunflower oil (MIX). The data were analyzed at 30 and 60 days in milk (DIM). Oil supplementation did not affect total dry matter intake or forage intake. The treatment SUN reduced daily milk yield, 4% fat-corrected milk yield, and milk fat, while the other treatments generated similar results. The treatment SUN reduced the milk net energy and the use efficiency of the NE_L for milk production and BW and BCS changes, and improved EB up to 60 DIM. On tropical pastures, supplementation with unsaturated oils for cows in early lactation does not result in better milk performance. The supplementation with sunflower oil improves the energy balance in early lactation.     

Genetic diversity of Mycoplasma hyopneumoniae isolates of abattoir pigs

Mycoplasma hyopneumoniae, the causative agent of porcine enzootic pneumonia, is present in swine herds worldwide. However, there is little information on strains infecting herds in Canada. A total of 160 swine lungs with lesions suggestive of enzootic pneumonia originating from 48 different farms were recovered from two slaughterhouses and submitted for gross pathology. The pneumonic lesion scores ranged from 2% to 84%.Eighty nine percent of the lungs (143/160) were positive for M. hyopneumoniae by real-time PCR whereas 10% (16/160) and 8.8% (14/160) were positive by PCR for M. hyorhinis and M. flocculare, respectively. By culture, only 6% of the samples were positive for M. hyopneumoniae (10/160). Among the selected M. hyopneumoniae-positive lungs (n = 25), 9 lungs were co-infected with M. hyorhinis, 9 lungs with PCV2, 2 lungs with PRRSV, 12 lungs with S. suis and 10 lungs with P. multocida. MLVA and PCR-RFLP clustering of M. hyopneumoniae revealed that analyzed strains were distributed among three and five clusters respectively, regardless of severity of lesions, indicating that no cluster is associated with virulence. However, strains missing a specific MLVA locus showed significantly less severe lesions and lower numbers of bacteria. MLVA and PCR-RFLP analyses also showed a high diversity among field isolates of M. hyopneumoniae with a greater homogeneity within the same herd. Almost half of the field isolates presented less than 55% homology with selected vaccine and reference strains.     

Distribution of glyphosate-resistant and glyphosate-susceptible hairy fleabane (Conyza bonariensis) in central California and their phenological development

Glyphosate-resistant (GR) hairy fleabane was first documented in California in 2007. A preliminary study comparing plants from this population to a glyphosate-susceptible (GS) population determined that they developed more rapidly than the GS plants. However, it was not known if this was a general characteristic of all GR populations. Therefore, seeds of hairy fleabane were collected from 122 locations in central California. Plants from each location were grown and tested for resistance using a shikimate enzyme assay. Seeds from 90 locations germinated of which 27 % were determined as GR, 21 % as GS, and 52 % as intermediate/mixed. Plants from five GR and five GS populations, as determined by the assay, were randomly selected for comparisons and were grown in 2010/2011. Time taken by each plant to reach key phenological stages was recorded and converted into growing degree day (GDD) using a base temperature of 4.2 °C. The biomass of each plant was recorded at seed set. The five GR and GS plants had similar phenological development rates and biomass. Therefore, the differences previously identified between the two populations were not present in the additionally tested populations. The phenological development was hence described by a common GDD model with the plants requiring 374, 711, 970, 1289, and 1551 GDD, to reach the rosette, bolting, first bud, first flower, and first seed stages, respectively. This study shows widespread distribution of GR hairy fleabane in central California, provides useful information on biology of this weed for management, and suggests possible label changes to herbicide manufacturers.     

Long-term mild hypoxia does not reduce thermal tolerance or performance of the freshwater prawn Macrobrachium tenellum

The present study was designed to assess the performance of the freshwater prawn Macrobrachium tenellum in optimal and sub-optimal dissolved oxygen conditions, considering increasing environmental pressures. Thermal tolerance and thermal metabolic scope (TMS) with related integrated biomarker response (IBR) were measured in prawns exposed to normoxia (80% air saturation), mild (40% air saturation) and severe hypoxia (25% air saturation) at three exposure time points (10, 20 and 30 days). Effects of hypoxia on thermal tolerance were not detectable over time; they were perhaps masked by hyperventilation, or by an increase or diversion of haemolymph processes. After 30 days, TMS was 11% higher in mild hypoxia compared with normoxia, while it was 64% lower in severe hypoxia, indicating the loss of aerobic metabolism capacity during the latter. Mild-hypoxia prawns maintained a high IBR over time, supported by antioxidant enzyme activities (mainly superoxide dismutase), which helped avoid the serious oxidative damage (proteins and lipids) seen in severe hypoxia animals, as well as lower acetylcholinesterase activity that indicated failure of communication between the nervous and locomotor systems. Our results documented a high tolerance by M. tenellum to mild-hypoxia events, which should be further tested under seasonal and extreme habitat/tank temperatures.     

Concentrate: forage ratio in the diet of dairy cows does not alter milk physical attributes

This study aimed to evaluate the effect of concentrate-to-forage ratio (C:F) on the performance, blood profile, and milk physicochemical characteristics of mid-lactation cows fed a corn silage-based diet. Twenty four Holstein cows, with BW 575?±?70 kg, body condition score (BCS) 3.1?±?0.2, milk yield 18.4?±?3.0 kg, and days in milk (DIM) 121?±?21 were randomly allocated into three treatments with C:F ratios of 35:65, 45:55, and 55:45 on a dry matter (DM) basis. Data was submitted to analyses of variance and regression. Increasing C:F from 35 to 55 % linearly enhanced milk production (22 to 23.6 kg day^?1) and serum urea nitrogen (16.8 to 19.6 mg/dL), while it linearly reduced lactose and fat in milk (4.8 to 4.6 %; 3.9 to 3.6 %, respectively). Body weight, BCS, milk acidity, ethanol stability, coagulation time, and milk and blood mineral contents did not differ among treatments. During the last period of measurements, increased C:F reduced urinary pH and milk urea nitrogen. Changes of concentrate-to-forage proportion from 35 to 55 % increased milk yield, altered chemical composition without changing BW, BCS, acidity, stability, and mineral content of milk and blood attributes.     

Replacement of wheat bran with spineless cactus (Opuntia ficus indica Mill cv Gigante) and urea in the diets of Holstein x Gyr heifers

The aim of this study was to evaluate the replacement effect of wheat bran with spineless cactus and urea in heifers. Twenty-four heifers with an average initial weight of 185?±?13 kg were used in this experiment. Four levels of spineless cactus corrected with urea and ammonium sulfate (9:1) were studied: 0, 33, 66, and 100 % replacement with wheat bran. Samples of feed, orts, and feces were analyzed to estimate the intake and digestibility of dry matter (DM) and nutrients. Indigestible neutral detergent fiber was used as an internal marker. The experiment was conducted in a completely randomized design. Dry matter, neutral detergent fiber, and total digestible nutrient intake demonstrated a quadratic effect (P?P?P?相似文献