Comparison of enzyme-linked immunosorbent assay and RT-PCR for the detection of porcine epidemic diarrhoea virus

Porcine epidemic diarrhoea (PED) is a contagious enteric disease of pigs caused by a coronavirus. A double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) based on the use of monoclonal antibodies was developed for the detection of porcine epidemic diarrhoea virus (PEDV). The DAS-ELISA was compared with RT-PCR in the examination of 506 specimens collected during 2006-2007 from pigs originating from different farms located in the Po valley. Both faecal samples obtained directly from the rectum of live animals showing clinical signs and intestinal samples collected from the caecum of deceased pigs were included in the study. The correlation between the two methods was higher when testing faecal samples (K = 0.97, 95% CI: 0.94-1.00) than testing intestinal samples (K = 0.62, 95% CI: 0.35-0.89). The use of ELISA technology provided an efficient and effective mean of evaluating the presence of coronavirus PED antigen in field samples and indicates that this procedure is a very useful tool in epidemiological studies.     

Antimicrobial susceptibility of clinical strains of Streptococcus suis isolated from pigs in Spain

The antimicrobial susceptibility of 151 clinical Streptococcus suis strains isolated from diseased pigs in Spain was determined by a microdilution method. Isolates were mostly susceptible to beta-lactam antimicrobials, aminoglycosides, enrofloxacin, novobiocin and spectinomycin. More than 87% of the S. suis isolates were resistant to tetracyclines, sulphonamides, macrolides and clindamycin. Strains of serotype 9 were significantly more resistant than strains of serotype 2 (P<0.05) to tylosin (94% versus 77%) and clindamycin (94% versus 64%). Eighty-seven percent of the S. suis isolates were resistant to at least four antimicrobials and nine isolates (6%) were resistant to at least six antimicrobials. The most frequently identified multidrug pattern involved resistance against tetracyclines, sulphonamides, macrolides and lincosamides, with 69% of the isolates exhibiting this resistotype. Fifteen out of the 22 strains of serotype 2 (68.2%), and 84 out of the 98 of the strains of serotype 9 (85.7%) exhibited this resistotype, indicating its widespread distribution among the strains of the two most frequently isolated serotypes.     

Local and systemic immune responses to Echinococcus granulosus in experimentally infected dogs

Local and systemic immune responses were studied in six dogs experimentally infected with the dog/sheep tapeworm Echinococcus granulosus. All dogs developed similar IgG antibody response to parasite antigens. In contrast, IgE and IgA responses differed widely. No relationship between IgA responses and parasite burden at the end of the infection were observed. Further, clear differences in the anti-parasite IgA response in serum as compared with specific IgA forming cells in mesenteric lymph nodes were observed within the same dog. An inverse association of anti-parasite IgE and parasite load seemed to be present, with the strongest IgE response in the one dog that had no worms in the intestine at the end of the experiment. No differences were observed in the numbers of intestinal mast cells and goblet cells among all infected dogs. However, the dog with no detectable parasite load had a marked reduction of detected mast cells in the submuscular and muscular layer of the mucosa. Our data give new insight into the immune response of dogs during E. granulosus infection and provide information that may be useful for the rational design of vaccines for the control of hydatid disease.     

Histochemical skeletal muscle fibre types in the sheep

In this study, the differentiation of adult and postnatal muscle fibres in sheep longissimus thoracis muscle has been characterized. By using a variety of histochemical methods, we have investigated the m-ATPase and metabolic activities of skeletal muscle fibres in adult sheep and lambs aged between 1 day and 3 months. Types I, IIA, IIB and IIC fibres were identified. The results showed that the interpretation of the fibre type composition depends on the methods used. The findings also revealed that the fibre types IIA and IIB can be separated histochemically in sheep by using the correct m-ATPase technique, even at early stages of postnatal development, and that the origin of the four different fibres of the adult can be traced back to early postnatal stages.     

Detection of Citrus Leaf Blotch Virus Using Digoxigenin-Labeled cDNA Probes and RT–PCR

Citrus leaf blotch virus (CLBV) was detected by dot-blot hybridization (DBH), and tissue print hybridization (TPH) and by one-step RT–PCR in citrus plants growing both in the greenhouse and in the field. DBH with digoxigenin-labeled cDNA probes allowed CLBV detection in dsRNA-rich and total RNA preparations equivalent to 5 and 0.1mg of infected tissue, respectively. DBH gave intense signals with RNA extracts from young bark, tender shoots and young leaves, whereas the best hybridization signals with TPH were obtained using tender shoots and young leaf petioles. One-step RT–PCR was 10-fold more sensitive than DBH and amplification was obtained with all infected tissues. CLBV was readily detected in young leaves of infected Eureka lemon, Marsh grapefruit, Nules clementine, Navelina orange and Nagami kumquat in the greenhouse, using either hybridization or RT–PCR, but not in leaves of Pineapple sweet orange. Detection in field trees was less consistent and was only achieved by RT–PCR and DBH. CLBV was detected by DBH and RT–PCR in different citrus varieties from several geographic areas showing bud union crease on trifoliate rootstocks, but not in neighbor trees with the same symptoms or in other varieties showing bud union crease on those rootstocks. Failure to detect CLBV in trees with bud union crease could be due to low virus titer or uneven distribution within the plant. Alternatively, a different agent could be involved in causing bud union crease.     

Nitric oxide synthase expression in the endothelium of pulmonary arterioles in normal and pulmonary hypertensive chickens subjected to chronic hypobaric hypoxia

To determine whether or not exposure to chronic hypoxia and subsequent development of pulmonary hypertension syndrome (PHS) induce alterations in endothelial nitric oxide (NO) production in broiler's pulmonary vascular bed of broilers, we studied the expression of nitric oxide synthase enzyme in pulmonary endothelial cells by a nicotinamide adenine dinucleotide phosphate (NADPH)-diaphorase histochemical staining reaction. For this purpose, 60 broilers of three different ages (17, 30, and 42 days) were used. The animals were distributed in two groups: a) 30 healthy (nonhypertensive) broilers and b) 30 chicks with PHS. All broilers in group b had fewer NADPH-diaphorase-positive endothelial cells in arterioles than did the nonhypertensive broilers. These differences were highly significant (P < 0.01). These results demonstrate for, the first time in broilers, that hypoxia-induced pulmonary hypertension is associated with a decrease of endothelial-derived NO expression in pulmonary vessels.     

Brucellosis in Central America

Brucellosis is a disease of domestic animals and humans in Central America (CA). Bovine and swine brucellosis caused by Brucella abortus and Brucella suis, respectively, have been identified in all CA countries, while ovine and caprine brucellosis caused by Brucella melitensis has been detected in Guatemala. The prevalence of bovine brucellosis is estimated between 4 and 8%, with higher prevalence in dairy herds, with losses calculated at US$ 25 million per year. National Control Programs based in calf vaccination and removal of the reactors have had little impact in the control of brucellosis in CA. In a region where experimentation with new vaccines is not affordable, unrestricted adult vaccination by the conjunctival route with S19 is recommended. This strategy is expected to reduce the prevalence and density of the bacteria to numbers where “clean” vaccination would be possible. Thereafter, serological identification and elimination of the reactors could be initiated under more favorable conditions of herd infection.     

Genetics of Six Components of Autofertility in Vicia faba

The inheritance of six components of autofertility was studied in faba beans in two consecutive years (1989 and 1990) by analysing parents, Fl and segregating generations of 14 crosses between preselected pure lines. Genetic analysis was performed by a joint scaling test proposed by Cavalli (1952). Additive gene effects were more important than dominance effects in flowers/node, pods/flower and ovules/ovary. In contrast, characters directly related with the seed such as pods/node, seeds/pod and, to a lesser extent, seeds/ovule showed high positive values of dominance. The three latter traits in general exhibited strong heterosis, probably due to genetic interactions. In order to produce selection gain, any breeding programme has to consider that dominance and heterosis vary according to the character involved. The present study reveals different degrees of dominance expression depending on cross specificity as well as on environmental conditions. The advantages of studying segregating generations instead of diallel crosses and the need for a simultaneous study of many crosses are emphasized in this paper.     

Use of neuromuscular blockade with rocuronium bromide for intubation in cats

ObjectiveTo determine whether neuromuscular blockade with rocuronium bromide (RB) would improve endotracheal intubation (EI) conditions in comparison with topical lidocaine hydrochloride (LH).Study designRandomized prospective study.AnimalsForty seven healthy cats of unspecified breed, aged 17 ± 11 months and weighing 2.8 ± 0.8 kg, undergoing elective procedures.MethodsAnesthesia was induced with xylazine (XZ) (1.1 mg kg^?1 IM) and tiletamine‐zolazepam (XTZ) (7 mg kg^?1 IM) and EI was attempted. Cats which could not be intubated at the first attempt (n = 34), were randomly medicated with either 0.1 mL LH 10% spray on the laryngeal mucosa (n = 17) or 0.6 mg kg^?1 intravenous RB (n = 17). Sixty seconds later, a second attempt at EI was performed. The effect of both drugs was assessed using a previously published scale (Sandor Agoston). EI conditions associated with laryngoscopy, vocal cord position and movement, cough, patient movement, time and attempts needed in order to perform EI were recorded. Heart rate and end‐expired CO₂ concentration were monitored.ResultsGroups were comparable in age, weight, gender and hematological parameters. Clinically acceptable EI conditions were not significantly different between RB and LH assisted groups (p = 0.31). However, there was a significant difference in cough, vocal cord movement and position between the RB and the LH groups. The group intubated at the first attempt and receiving neither RB nor LH coughed persistently (11/13). The cats receiving RB had to be ventilated for 10–28 minutes.Conclusions and clinical relevanceThe present study shows that, when used in cats anesthetized with XTZ, RB paralyzes the internal laryngeal muscles keeping the vocal cords in an intermediate position (paramedial) 60 seconds after being administered. RB is an effective alternative to LH to overcome the airway protective reflexes when performing EI but requires ventilatory support until the paralysis wears off.