In vitro inhibition of Citrobacter freundii, a red-leg syndrome associated pathogen in raniculture, by indigenous Lactococcus lactis CRL 1584

Red-leg syndrome (RLS) is one of the main infectious diseases that cause economic losses in Lithobates catesbeianus hatcheries, Citrobacter freundii being an etiological agent. Treatment or prevention with therapeutics or chemicals results in modifications of the indigenous microbiota, development of antibiotic resistance, presence of their residues in food and enhancement of production costs. Thus, probiotics could be used as an alternative therapy. Lactic acid bacteria are part of the indigenous microbiota of healthy frogs and can prevent pathogen colonization by different mechanisms, including the production of antagonistic substances. In this work, the evaluation and characterization of the inhibition of C. freundii CFb by Lactococcus lactis subsp. lactis CRL 1584, a potentially probiotic candidate, were carried out. This strain produced lactic acid, H(2)O(2) and bacteriocin in static and shaken conditions and inhibited pathogen growth in associative cultures, with an earlier inhibition under agitated conditions. The elimination of each of the antimicrobial metabolites partially abolished the inhibition of the pathogen, suggesting that the inhibitory effect could be attributed to a combined action of the three antagonistic molecules. Electron microphotographs revealed the damage caused by L. lactis CRL 1584 supernatants to C. freundii cells. The addition of pure lactic acid, H(2)O(2) and bacteriocin to the culture media showed that each metabolite caused different morphological modifications in C. freundii, in agreement with the effect on viable cell counts. The results support the possibility that L. lactis CRL 1584 might be considered as a probiotic to be used in the prevention of RLS in raniculture.     

Assessment of stakeholder perceptions of the damage to silo bags by vertebrate species in Argentina

We conducted an on-line questionnaire to assess stakeholder perceptions about damage to silo bags by vertebrate species in Argentina. We received 190 responses reporting damage to silo bags by vertebrates from the Pampas, Espinal, and Chaco regions. More than two-thirds of respondents indicated armadillos were the main harmful species, followed by rodents and dogs. Frequency of damage to silo bags was higher for armadillos (79%) than for the other species together (64%), and was higher in the Pampas (79%) than in the other regions (45–66%), suggesting a latitudinal gradient of damage. Economic losses were reported as “medium” or “high” by 37% of respondents. According to these stakeholders, damage to silo bags by vertebrates is a regionally extended phenomenon in most of the Argentine agricultural region and it is recognized as economically relevant by these stakeholders. Management of this conflict should focus on environmental aspects.     

Evaluation of transformation growth factor beta1, interleukin-10, and interferon-gamma in male symptomatic and asymptomatic dogs naturally infected by Leishmania (Leishmania) chagasi

The aims of this study were to evaluate the immunomodulatory role of TGF-β₁, IL-10, and INF-γ in spleen and liver extracts and supernatant cultures of white spleen cells from male symptomatic and asymptomatic dogs, naturally infected by Leishmania (Leishmania) chagasi. Thirty dogs from Araçatuba, São Paulo, Brazil, an endemic leishmaniosis area, were selected by positive ELISA serological reaction for Leishmania sp. and divided into two groups: asymptomatic (n = 15) and symptomatic (n = 15) consisting of animals with at least three characteristic signs (fever, dermatitis, lymphoadenopathy, onychogryphosis, weight loss, cachexia, locomotion problems, conjunctivitis, epistaxis, hepatosplenomegaly, edema, and apathy). After euthanasia, spleen and liver fragments were collected for ex vivo quantification of TGF-β₁, IL-10, and INF-γ. Naturally active in vitro produced TGF-β₁ was also evaluated in spleen cell culture supernatant. Spleen and liver extract of asymptomatic dogs had higher mean TGF-β₁ levels than symptomatic dogs. High concentrations of IL-10 were found in spleen, and mainly in liver extract of both groups. Higher INF-γ concentrations were found in spleen extracts of symptomatic dogs, and in liver extracts of asymptomatic dogs. Extract of this cytokine was lower in spleen extract. Although INF-γ is being produced in canine infection, mean levels of TGF-β₁ and IL-10 from spleen and liver extracts were quantitatively much higher; suggesting that immune response in both asymptomatic and symptomatic dogs was predominantly type Th2.     

Delayed acute capture myopathy in three roe deer

Delayed acute capture myopathy is the term used to describe the clinical syndrome observed in three roe deer captured by drive-nets and transported to an enclosure for scientific purposes. The animals died 48 h, 60 h and 8 days after being captured. The simultaneous deaths coincided with a previous episode of deliberate human disturbance. The histopathological findings were indicative of acute myopathy and myoglobinaemic nephrosis. These could be related to an ataxic myoglobinuric syndrome brought on by capture and transport operations. The lack of clinical signs and negative prognosis indicators in the period between capture and just before death. the absence of gross muscular lesions in the animal that died after 8 days post-capture, the simultaneous deaths of animals captured at different times and the evidence of deliberate human disturbance in the enclosure are suggestive of death triggered by a second stress episode.     

In vitro susceptibility of field isolates of Francisella tularensis subsp. holarctica recovered in Spain to several antimicrobial agents

Forty-two recent (1997-1999) Spanish isolates of Francisella tularensis subsp.holarctica were tested in a broth microdilution method for their susceptibilities to 29 antimicrobial agents, including penicillins, cephalosporins, cephamicins, monobactams, penems, aminoglycosides, tetracyclines, macrolides, quinolones, chloramphenicol and fosfomycin. The isolates were resistant to beta-lactam antibiotics and susceptible to chloramphenicol, ciprofloxacin, levofloxacin and norfloxacin.     

Comparison of two automated spectrophotometric methods for ceruloplasmin measurement in pigs

Ceruloplasmin (Cp) determination could provide an objective measure of the health status of an animal and could be used as marker of animal health and welfare (Skinner, 2001) (Skinner, J. G., 2001. Special report. International standardization of acute phase proteins. Veterinary Clinical Pathology 30, 2-7.) but only manual methods have been reported to determine Cp concentrations in pigs (McCosker, 1961; Toussaint et al., 1995; Eckersall et al., 1996) (McCosker, P. J., 1961. Paraphenylenediamine oxidase activity and copper-levels in mammalian plasmas. Nature. 190, 887-889; Toussaint, M. J. M., Van Ederen, A. M., Gruys, E., 1995. Implication of clinical pathology in assessment of animal health and in animal production and meat inspection. Comparative Haematology International 5, 149-157; Eckersall, P. D., Saini, P. K., McComb, C., 1996. The acute phase response of acid soluble glycoprotein, alpha(1)-acid glycoprotein, ceruloplasmin, haptoglobin and C-reactive protein, in the pig. Veterinary Immunology and Immunopathology 51, 377-385). In the present study two automated methods based on the use of two different substrates for the determination of serum ceruloplasmin in pigs were developed, evaluated and compared. Both methods showed a good precision and detection limits, with no signs of inaccuracy and could be applied to biochemical autoanalyzers usually found in clinical laboratories using only minimal amounts of serum. Additionally the behaviour of Cp in experimental and clinical situations was studied showing an increase of around two fold after turpentine administration and significantly higher values in cases of pigs with inflammatory conditions when compared with healthy pigs.     

Evaluation of an antigen capture ELISA for the early diagnosis of Hypoderma lineatum in cattle under field conditions

An antigen capture or sandwich ELISA (sELISA) was evaluated for the diagnosis of Hypoderma lineatum in cattle under field conditions in northwestern Spain. The kinetics of circulating hypodermin C (HyC) and specific antibodies during the course of natural infestation were determined in a group of 10 Frisian calves. In addition, oesophagi and blood samples were taken from 105 cows at a slaughterhouse in order to compare three methods for the diagnosis of H. lineatum: sandwich ELISA for the detection of the antigen HyC (sELISA), indirect ELISA for the detection of antibodies anti-HyC (iELISA) and the detection of first instars (L1) in the oesophagus. In naturally infested cattle, HyC was present in circulation at low levels during the early and late phases of the infestation. However, in the middle phase, coinciding with the presence of L1 in the oesophagus, two peaks of increased HyC concentration were observed. Specific antibodies increased progressively until the first appearance of larvae in warbles on the back. There was no correlation between antigen or antibody levels and the number of grubs in the back. Prevalence of first instars in the oesophagi of slaughtered cows was 21.9% (23/105). The percentage of cattle that were positive for circulating antigen was slightly higher (24.8%), suggesting the recent destruction of migrating larvae in some animals. However, there was no correlation between the number of L1 and HyC levels. With the iELISA, 79% of the animals were positive to Hypoderma, which means that a high percentage of those animals have been exposed to the parasite but they had no apparent current infestation. The sELISA is a good tool to follow larval development within the host; however, the episodic elevation of HyC levels limits the usefulness of this test for the early diagnosis of Hypoderma under field conditions.