Genetic variability of avian Escherichia coli strains evaluated by enterobacterial repetitive intergenic consensus and repetitive extragenic palindromic polymerase chain reaction

In this study, we tested the capability of enterobacterial repetitive intergenic consensus (ERIC) and repetitive extragenic palindromic (REP) polymerase chain reaction (PCR) to detect genetic diversity among Escherichia coli strains isolated from chickens bearing clinical signs of colibacillosis and compared the genotypes so obtained with the O:H serotypes and virulence of those strains. The DNAs from 50 avian E. coli strains and from E. coli ATCC 25922 were used to amplify ERIC and REP sequences. DNA from avian strains produced from 8 to 17 bands by ERIC-PCR and from 6 to 20 bands by REP-PCR; E. coli ATCC produced 11 bands by both methods. ERIC and REP-PCR showed good discriminating power, and the dendograms based on the different patterns revealed extensive genetic diversity among the avian strains. Those strains were allocated into four major clonal clusters, each one with 60% of similarity by ERIC and REP-PCR, and those clusters corresponded to strains with different degrees of pathogenicity. However, 56% of the pathogenic strains (28/50) belonged to two out of three major clonal clusters, and 86% of the nonpathogenic strains tended to group in one cluster and one subgroup. The 32 serotypes detected were distributed in all clusters, and within a serogroup, different DNA fingerprints were observed; however, strains with same serotypes tended to form clusters with similarity coefficients greater than 80%. These results suggest that no specific serotype and genotype is responsible for colibacillosis and that REP and ERIC-PCR are reproducible techniques that can improve the studies needed to clarify the pathways to the pathogenesis of colibacillosis.     

Efficacy of a long-acting formulation of ivermectin against Psoroptes ovis (Hering, 1838) on cattle

A study was conducted in cattle experimentally infested with Psoroptes ovis to compare the prophylactic control against P. ovis provided by a long-acting injectable formulation of ivermectin to that of a commercially available injectable formulation of doramectin. Thirty Holstein steers were used. Animals were allocated by restricted randomization based on Day 0 body weight, forming six replicates of five animals each. Within each replicate, one animal was randomly allocated to one of the following treatment groups, with ivermectin and doramectin administered subcutaneously where indicated: (1) untreated controls; (2) ivermectin long-acting injectable (LAI) 630 mcg/kg, 56 days before challenge; (3) ivermectin LAI 630 mcg/kg, 42 days before challenge; (4) ivermectin LAI 630 mcg/kg, 35 days before challenge; or (5) doramectin 200 mcg/kg, 35 days before challenge. Animals were housed in individual pens 1 week prior to treatment. All animals were experimentally infested with P. ovis mites in the area between the shoulders, on the same day. Live mites were counted in scrapings from mange lesions at 2 sites on each animal 14, 21 and 28 days after challenge. Live mites were found in 33, 67 and 83% of the untreated controls on each respective evaluation. No P. ovis mites were found in steers treated with ivermectin LAI. Those animals showed lower (P < 0.05) mite counts than untreated controls on evaluations conducted 21 and 28 days after challenge. These results indicate that the ivermectin long-acting injectable formulation prevents induced infestations by P. ovis for at least 56 days after treatment. Doramectin injectable formulation, used at 200 mcg/kg, did not have a prophylactic effect 35 days after treatment.     

Congenital malformations of the tricuspid valve in domestic carnivores: a retrospective study of 50 cases

A retrospective study was done on 14 cats and 36 dogs diagnosed with right atrioventricular valve malformations (RAVM), either tricuspid dysplasia (n = 38) or Ebstein's anomaly (n = 12). Comparison with a large reference population including 85 250 animals allowed an epidemiologic analysis of these RAVM, demonstrating that Labrador, Boxer and Chartreux cats had a 35, 7 and 11-fold higher risk respectively to be affected by one of these heart diseases than other breeds, without sexual predisposition. In 93% of cases, cardiac auscultation revealed a heart murmur, whose grade was not significantly correlated with the echographic stages of the valvular disease. The echocardiographic examination confirmed the frequent association between RAVM and other congenital anomalies, particularly mitral dysplasia, in 84% and 20% of tricuspid dysplasia and Ebstein anomaly respectively. Last, the prognosis of RAVM was worsened by atrial fibrillation and right-sided heart failure.     

Recombinant single-chain canine interleukin 12 induces interferon gamma mRNA expression in peripheral blood mononuclear cells of dogs with visceral leishmaniasis

Canine visceral leishmaniasis poses important concerns for public health and veterinary medicine in many areas of the world. Resistance to it seems to be associated with cellular specific immune responses of the so-called Th1 type. Interleukin-12 (IL-12) is one of the most potent inducers of Th1 type of immune responses to co-administered antigens. Herein, the cloning of canine IL-12, as a single-chain fusion protein (sccaIL-12), and its expression in biologically active form in COS-7 cells is reported. Supernatants from these cells stimulated the expression of comparable amounts of interferon gamma mRNA in peripheral blood mononuclear cells from dogs with natural visceral leishmaniasis. In addition, after stimulation with sccaIL-12, there was no difference between interferon gamma mRNA expressions in peripheral blood mononuclear cells of dogs with visceral leishmaniasis and from normal healthy control animals.     

B-mode and M-mode echocardiography of endurance horses raised in São Paulo State, Brazil

This study aimed to evaluate cardiac dimensions and indexes of cardiac function in endurance horses using bidimensional- (B-) mode and guided unidimensional- (M-) mode echocardiography. Thirty-five Arabian and crossbred Arabian male and female horses, aged 5 to 18 years old, with an average weight of 415.51 ± 36.76 kg and at least 1 year of endurance training and competition experience, were used. The following values were obtained for B-mode echocardiography in the right parasternal window (see “Materials and Methods” for expanded terms): RVIDd 2.54 ± 0.56 cm; RVIDs 2.08 ± 0.68 cm; IVSd 2.60 ± 0.31 cm; IVSs 3.90 ± 0.44 cm; LVIDd 10.07 ± 0.85 cm; LVIDs 6.65 ± 0.92 cm; LVFWd 2.50 ± 0.30 cm; LVFWs 3.74 ± 0.35 cm; Aod 7.86 ± 0.72 cm; LAs 10.84 ± 0.93 cm; Vd 578.40 ± 108 cm³; Vs 232.68 ± 70.61 cm³; FS% 34.99% ± 8.02%; EF% 60.13% ± 8.29%; E-point 2.25 ± 0.49 cm; and LAs:Aod 1.38 ± 0.16 cm. Using guided M-mode, the following values were obtained: RVIDd 2.04 ± 0.55 cm; RVIDs 1.61 ± 0.71 cm; IVSd 2.68 ± 0.29 cm; IVSs 4.17 ± 0.42 cm; LVIDd 9.72 ± 0.72 cm; LVIDs 5.94 ± 0.96 cm; LVFWd 2.69 ± 0.32 cm; LVFWs 4.23 ± 0.69 cm; Aod 7.21 ± 0.75 cm; Vd 534.24 ± 87.53 cm³; Vs 181.75 ± 67.69 cm³; FS% 39.06% ± 7.66%; EF% 66.38% ± 9.41%; and E-point 1.96 ± 0.58 cm.

Introduction

In equine sports medicine, it is important to evaluate the cardiovascular condition of the athlete. More recently, echocardiography has been used for this purpose.Echocardiography is an invaluable diagnostic aid, since it is non-invasive and easy to perform,[1, 2 and 3] while offering a dynamic and functional evaluation of the heart. [2, 3, 4 and 5].Initially, studies on the M-mode were made, including measurements of normal cardiac dimensions in horses[6 and 7] and foals and for the diagnosis of several cardiac diseases. [1] With the introduction of the B-mode technique in the mid-1980s and its validation, a more detailed anatomic evaluation was possible, [2, 8 and 9] thus allowing the observation of cardiac abnormalities. [10]Though echocardiographic examination could be used to predict the athletic condition of individuals,[11] this is possible only if normal echocardiographic reference values are established. As different breeds have different biotypes and develop different aptitudes, it is necessary to provide enough information for each type of work or breed. Research includes data for the Thoroughbred racehorse [3, 12 and 13] and Standardbred trotter, [14 and 15] which perform essentially anaerobic work.The endurance race is a modality that is growing significantly in Brazil, both in number of participants and in quality and training of the animals. The endurance horse demands a physical conditioning that differs from that required for racehorses, especially owing to the development of aerobic work. In addition, the animals are mainly Arabian or Arabian crossbred, which have a particular biotype. Because of these facts, and the lack of data about the echocardiographic behavior of endurance horses, this study aimed to determine the dimensions and indexes of cardiac function for horses with endurance training using B-mode and guided M-mode echocardiography.

Materials and methods

Thirty-five clinically normal endurance Arabian and Arabian crossbred male and female horses between 5 and 18 years old, with an average weight of 415.51 ± 36.76 kg, were included in the study. All animals had at least 1 year of training and experience in competitions and were kept under similar hygienic, sanitary, and nutritional management. Training consisted of trail rides, alternating walking, trotting, and galloping, from 20 to 40 km per day, 3 to 4 times per week.The echocardiographic examination was performed with a Sonosite model 180 Plus v. 1.9 echocardiograph, using a 2-4–MHz electronic microconvex transducer with a maximal depth of 24 cm.The animals were prepared as previously reported,[4, 16 and 17] the hair being coated with large amounts of acoustic coupling gel. During the examination, the animals were at rest, calm, and physically restrained only by the halter, in a quiet and dark room.The following echocardiographic measurements, in B-mode as well as guided M-mode, were performed: the left atrium internal diameter in systole (LAs), only in B-mode, in the long axis view, dorsal to the annulus[15]; the right ventricle internal diameter, in diastole (RVIDd) and in systole (RVIDs); the interventricular septum thickness, in diastole (IVSd) and in systole (IVSs); the left ventricle internal diameter, in diastole (LVIDd) and in systole (LVIDs); and the left ventricle free wall thickness, in diastole (LVFWd) and in systole (LVFWs), from the left ventricle short axis view at the chordal level; and the aortic root internal diameter (Aod), from the aortic valve short axis view. The B-mode measurements were taken according to Patteson et al, [12] and the M-mode measurements were based on Long et al. [18]Images were obtained from the right hemithorax. The position used for the transducer to obtain the standardized images for B-mode and guided M-mode were determined and used as described in Reef.[4 and 17]The echocardiographic examination began using the B-mode echocardiography, with the establishment of the tomographic planes, evaluating morphology of the cardiac structures and their movement in a global manner.The first image obtained was the right parasternal long axis view, with the observation of the ventricle inlets, atrioventricular valves, and both atria. With the transducer positioned to optimize visualization of the left atrium, usually with a minimal dorsal angulation, the LAs measurement was made, immediately before the opening of the mitral valve (Fig 1).     

Diagnostic potential of natriuretic peptides in the occult phase of golden retriever muscular dystrophy cardiomyopathy

The objective of the study was to determine whether the plasma concentrations of atrial and brain natriuretic peptides (ANP and BNP, respectively) could be reliable markers of cardiac alterations during occult cardiomyopathy in Golden Retriever Muscular Dystrophy (GRMD). Fifty Golden Retrievers without any clinical or radiographic sign of heart disease were included in this study (21 GRMD dogs and 29 controls). Controls and GRMD dogs were divided into 2 subgroups according to age (< and > or =12 months old, respectively). All dogs underwent echocardiography and determination of BNP and ANP plasma concentrations by radioimmunoassay. No ventricular dilatation or dysfunction was observed in either control or GRMD dogs. ANP plasma concentration did not differ significantly between controls and GRMD dogs (mean +/- SD = 72 +/- 49 versus 58 +/- 23 pg/mL, respectively, P = .21). This finding was confirmed in both subgroups of dogs (ie, those < and > or =12 months old). In contrast, BNP plasma concentrations were significantly higher in GRMD dogs than in controls (mean +/- SD = 117 +/- 92 versus 46 +/- 22 pg/mL, respectively, P < .05). In dogs > or =12 months old, sensitivity and specificity of BNP for identifying GRMD with a cutoff of 65 pg/mL were 78 and 86%, respectively. For the same cutoff value, sensitivity dropped to 42%, whereas specificity reached 100% in dogs <12 months old. In conclusion, BNP may be a useful biochemical marker of asymptomatic cardiomyopathy. However, this peptide does not allow very early detection because its optimal discriminatory power was observed in adult dogs (ie, > or =12 months of age).     

Ultrastructural Observations and DNA Degradation Analysis of Pepper Leaves Undergoing a Hypersensitive Reaction to Xanthomonas campestris pv. Vesicatoria

Ultrastructural details of the hypersensitive reaction induced by infiltration with avirulent race 2 Xanthomonas campestris pv. vesicatoria in pepper Early Calwonder-10R leaves (incompatible interaction) are reported. Affected cells displayed plasmalemma undulations and disruption, lysis of the chloroplast membrane, degeneration of other organelles, general cytoplasm disorganisation and, often, protoplast shrinkage. The nuclei contained large masses of electron-dense material, apparently formed by chromatin aggregation. In many cases a single chromatin-like layer was deposited on the inner side of the nuclear envelope leaving a finely granular matrix in the centre of the nucleus; the nucleolus usually disappeared. The nuclear envelope was sometimes ruptured and the internal matrix leaked into the cytoplasm. The content of many affected cells eventually coagulated and became very electron-dense. The walls often collapsed. All these alterations were especially visible in spongy mesophyll cells at sites where bacteria occurred in the intercellular spaces. Although some of the nuclear and cytoplasmic alterations recall certain aspects of apoptotic cell death, molecular determinations did not reveal any DNA degradation in hypersensitively reacting tissues. The first cell alterations in leaves infected with the virulent bacterial race 1 (compatible interaction) were observed only 27h after inoculation, when the cytoplasm of some cells showed limited internal disorganisation and plasmolysis at sites where bacterial colonies developed.     

A kinetic model of phosphorus metabolism in growing goats

The effect of increasing phosphorus (P) intake on P utilization was investigated in balance experiments using 12 Saanen goats, 4 to 5 mo of age and weighing 20 to 30 kg. The goats were given similar diets with various concentrations of P, and 32P was injected to trace the movement of P in the body. A P metabolism model with four pools was developed to compute P exchanges in the system. The results showed that P absorption, bone resorption, and excretion of urinary P and endogenous and fecal P all play a part in the homeostatic control of P. Endogenous fecal output was positively correlated to P intake (P < .01). Bone resorption of P was not influenced by intake of P, and P recycling from tissues to the blood pool was lesser for low P intake. Endogenous P loss occurred even in animals fed an inadequate P diet, resulting in a negative P balance. The extrapolated minimum endogenous loss in feces was .067 g of P/d. The minimum P intake for maintenance in Saanen goats was calculated to be .61 g of P/d or .055 g of P/(kg(.75) x d) at 25 kg BW. Model outputs indicate greater P flow from the blood pool to the gut and vice versa as P intake increased. Intake of P did not significantly affect P flow from bone and soft tissue to blood. The kinetic model and regressions could be used to estimate P requirement and the fate of P in goats and could also be extrapolated to both sheep and cattle.