Evaluation of the humoral and cellular immune response to different antigens of Corynebacterium pseudotuberculosis in Canindé goats and their potential protection against caseous lymphadenitis

Corynebacterium pseudotuberculosis is the etiologic agent of caseous lymphadenitis, a disease that affects goats and sheep, and can cause severe economic losses. In this study, four different antigenic extracts were obtained from the attenuated strain T1, which was isolated in the state of Bahia (Brazil). Forty-four Canindé breed goats were divided in five groups, each receiving a different antigen solution and saline buffer as a control. The humoral response was monitored through the identification of specific IgG by indirect ELISA and Western Blotting, and the production of IFN-gamma was followed in order to observe the activation of cellular response. After twelve weeks of antigen inoculation, the animals were challenged with 2 x 10(5)CFU of a wild strain, also isolated in Bahia, and necropsy was performed on all animals twelve weeks afterwards. It was observed that the attenuated bacteria gave a protection of 33.3%, in addition to the weak humoral response elicited. Animals inoculated with secreted antigen associated with Freund's incomplete adjuvant and oligodeoxynucleotide containing unmethylated CpG dinucleotides (CpG ODN) showed a strong humoral response, but this inoculation could not prevent the spread of challenge bacteria in the majority of animals. These results demonstrate the immunogenic potential of the attenuated T1 strain in the development of a vaccine against caseous lymphadenitis in goats.     

Growth and feed utilization of gilthead sea bream (<Emphasis Type="Italic">Sparus aurata</Emphasis>, L.) fed to satiation and restrictively at increasing dietary energy levels

Three isoproteic (47% protein) diets were formulated to contain graded levels of crude fat (diet D16:16%, diet D24: 24% and diet D32: 32%). Each diet was fed to satiation in three and to 80% satiation in two replicate groups of gilthead sea bream (Sparus aurata), having an initial body weight of 72–74 g. The trial lasted 81 days. Groups fed to satiation showed higher final body weight (FBW; 238.8–252.3 g vs. 218.0–229.3 g) and daily growth index (DGI; 2.49–2.65%/day vs. 2.27–2.34%/day) than those fed to 80% satiation. Feed intake was significantly different both for feeding level and for diet composition. Fish fed to satiation had higher feed conversion rate (FCR) compared to the 80% satiation groups (1.33–1.44 vs. 1.13–1.17; P ≤ 0.001). Within satiation groups, FCR was significantly lower in fish fed D16 compared to fish fed D32 (1.33 vs. 1.44, P ≤ 0.05), whereas no statistical differences were found within the 80% satiation groups. The increase in dietary lipid level did not improve growth performance, feed efficiency and protein utilization but decreased gross lipid efficiency. Conversely, a reduction in ration from satiation to 80% satiation decreased DGI, thus improving FCR. Feed costs were influenced by dietary energy level and feeding ratio, the lowest energy diet at 80% satiation being the most profitable combination among the variables.     

Changes in xyloglucan endotransglucosylase/hydrolase (XTHs) expression and XET activity during apple fruit infection by Penicillium expansum Link. A

Although the plant cell wall is an extremely effective physical barrier against attack by pathogens, most phytopathogenic microorganisms produce enzymes that are capable of degrading cell wall polymers such as pectin and hemicelluloses. In this work we investigate the possible implication of the cell wall enzyme XET in the host-pathogen interaction as well as the XTH gene regulation during apple fruit infection by Penicillium expansum Link. A. We characterized the time course of fungal infection in apple fruit, and the XET activity and the expression of eleven MdXTHs. The results showed an important decrease in XET specific activity in the fruits after 24, 48 and 72 h of fruit infection (42.9 %, 73.1 % and 95.1 % respectively as compared with the controls). The expression analysis of the MdXTHs showed an important decrease in expression, as the infection progressed, in MdXTH2 and MdXTH10, as well as in MdXTH3, MdXTH4 and MdXTH5, but mainly after 72 h of fruit infection; and in MdXTH1, and MdXTH9 after 24 and 48 h of infection. These results suggested that the decrease in XET activity during infection could be mainly attributed to the decrease in expression of MdXTH2 and MdXTH10, since those were the MdXTHs that showed the highest expression levels during ripening, particularly MdXTH10. These results support the suggestion that inhibition of MdXTHs expression, and consequently the decrease in XET specific activity, could represent a pathogenicity factor, since the potential reconstructing role of the enzyme also decreased and the infection could progress.     

Use of haloperidol and azaperone for stress control in roe deer (Capreolus capreolus) captured by means of drive-nets

The physical capture of wild ungulates is performed for different purposes when anaesthesia in field conditions is not possible or advisable. The use of tranquilizers may contribute to improved welfare of captured animals. We studied the effect of haloperidol and azaperone on the stress response of roe deer (Capreolus capreolus) through the study of physiological, haematological and serum biochemical parameters. Thirty one roe deer were drive-net captured and randomly injected with haloperidol (0.30 ± 0.04 mg/kg IM; n = 13), azaperone (0.43 ± 0.07 mg/kg IM; n = 11) or saline (0.5 mL IM; n = 7), and restrained for 3 h. The interindividual variability of heart rate was lower in the treated deer, suggesting a calming effect, and erythrocyte and biochemical parameters indicated vasodilation, splenic sequestration, hemodilution, improvement of renal perfusion and a protective effect on muscle. These results support the suitability of using either azaperone or haloperidol in capture operations of roe deer, in order to reduce stress and prevent its adverse effects.     

Continuous positive airway pressure administered via face mask in tranquilized dogs

Objective – To evaluate the tolerance of a continuous positive airway pressure (CPAP) mask in tranquilized dogs and compare PaO₂ in arterial blood in dogs receiving oxygen with a regular face mask or CPAP mask set to maintain a pressure of 2.5 or 5 cm H₂O. Design – Prospective, randomized clinical study. Setting – University teaching hospital. Animals – Sixteen client‐owned dogs without evidence of cardiopulmonary disease were studied. Interventions – Eight animals were randomly assigned to each of 2 treatment groups: group A received 2.5 cm H₂O CPAP and group B received 5 cm H₂O CPAP after first receiving oxygen (5 L/min) by a regular face mask. Animals were tranquilized with acepromazine 0.05 mg/kg, IV and morphine 0.2 mg/kg, IM. An arterial catheter was then placed to facilitate blood sampling for pHa, PaO₂, and PaCO₂ determinations before and after treatments. Direct mean arterial pressure, heart rate, respiratory rate, and temperature were also recorded after each treatment. Measurements and Main Results – CPAP administration was well tolerated by all animals. The mean arterial pressure, heart rate, respiratory rate, temperature, PaCO₂, and pHa, did not differ at any time point between groups. Differences were seen in oxygenation; in group A, PaO₂ significantly increased from a mean of 288.3±47.5 mm Hg with a standard mask to a mean of 390.3±65.5 mm Hg with the CPAP mask and in group B, PaO₂ increased similarly from 325.0±70.5 to 425.2±63.4 mm Hg (P<0.05); no differences were detected between the 2 CPAP treatments. Conclusions – In healthy tranquilized dogs noninvasive CPAP is well tolerated and increases PaO₂ above values obtained when using a regular face mask.     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

Neutrophilic myeloperoxidase index and mean light absorbance in neonatal septic and nonseptic foals

Background: Two neutrophilic indices reported by the ADVIA 120 Hematology Analyzer, neutrophilic myeloperoxidase index (MPXI), and mean light absorbance (neutrophil X mean [NXM]) have been proposed as indicators of systemic inflammatory disease in horses and of neutrophil activation in coronary ischemic syndromes in people. Objective: The aim of this study was to evaluate NXM and MPXI in healthy, sick nonseptic, and sick septic foals to determine whether conditions likely associated with neutrophil activation result in decreases in these variables. Methods: In this retrospective study, CBC data from 61 neonatal foals presented to the Equine Teaching Hospital of Barcelona were evaluated for correlations between MPXI, NXM, percentage of large unstained cells, neutrophil count, and percentage of band neutrophils. Results obtained in septic (n=32), sick nonseptic (n=22), and healthy foals (n=7) were compared. In addition, results recorded in septic/neutropenic (n=12), septic/non‐neutropenic (n=20), nonseptic/neutropenic (n=8), nonseptic/non‐neutropenic (n=14), and healthy foals (n=7) were also compared. Results: A weak negative correlation was found between MPXI and neutrophil count and between NXM and percentage of band neutrophils. Septic/neutropenic foals had significantly higher MPXI values (median 17.9, minimum–maximum 4.7–42.5) than did septic/non‐neutropenic (1.5, ?24.4 to 22.3), nonseptic/neutropenic (6.6, 0.6–17.9), and nonseptic/non‐neutropenic foals (8.8, ?10.1 to 16.8) but did not differ significantly from controls (12.8, ?8.5 to 20.4). Conclusions: Significant differences in NXM or MPXI were not found when disease groups were compared with controls; however, septic/neutropenic foals had significantly higher median MPXI than other groups of sick foals. Further prospective studies are needed to clarify if this finding is related to decreased neutrophil function or activation in septic/neutropenic foals.     

EARLY DIAGNOSIS OF BORON DEFICIENCY IN CHESTNUT

This study aims to identify an alternative plant tissue to be used in the early diagnosis of boron (B) deficiency in chestnut (Castanea sativa Mill.). A B-deficient orchard was selected, and 16 trees were submitted to two levels of B fertilization. When flowers were in bloom, the following tissues were sampled: leaves, androgynous catkins and flowers. There was a significant increase in B content in plant tissues due to B application. In July, the highest B content was observed in flowers in B0, but leaves had the greatest content in B1. Boron content in the tissues collected in July was positively correlated with B contents in leaves sampled in September. Foliar B concentrations, irrespective of the sampling period, were correlated with chestnut productivity, while the other tissues did not. These results suggest that the leaves, sampled in bloom, were the most efficient tissue for the early diagnosis of B deficiency.