Molecular characterization of Listeria monocytogenes and the significance for food hygiene

Research on the pathogen Listeria monocytogenes is a key issue both for the clinical and the food microbiologist owing to the unique pathway of infection and the exposure of humans via contaminated foods. Although, in Austria, the incidence of listeriosis is about 870-fold lower than the incidence for Salmonella infection, the food law manages both foodborne pathogens with a comparable stringency. The current risk management is based on the assumption that environmental L. monocytogenes isolates, from which the pool of "foodborne" isolates is recruited, are of similar pathogenicity compared to clinical and outbreak isolates. This verdict became doubted in the recent years. Characterization of L. monocytogenes by virulence gene sequencing, virulence studies in vivo and in vitro and by molecular typing was considerably stimulating the discussion on virulence variability in L. monocytogenes. This article provides insights in the value of epidemiological follow-up studies by presenting a typing study on 15 cases of listeriosis observed in a district hospital in Turkey. Furthermore results from typing L. monocytogenes either by virulence gene sequencing, mismatch amplification mutation assay or by pulsed field gel electrophoresis are discussed. The close interaction of molecular microbiology with food microbiology both in applied and basic science is currently creating a new discipline of molecular food microbiology. We are convinced that veterinary medicine will contribute to this exiting development in a fruitful way.     

An initial survey of the cattle grub Dermatobia hominis (L. Jr.) in Nicaragua

After the civil war and the Hurricane-Mitch disaster, cattlemen in Nicaragua were forced to transport their cattle from lowland areas to higher, dryer areas of the country. These areas are natural ecological niches for the cattle grub Dermatobia hominis (L. Jr.) (Diptera: Cuterebridae). To determine the importance of this infestation, the Agricultural and Livestock-Forestry Ministry selected a central area of Nicaragua to run a pioneer survey program to acquire information about hosts involved, number of cases, treatments applied and general knowledge of 42 farmers about the life cycle of the parasite. The subjects were either farm owners or farm managers. Ninety-five percentage of the farms indicated cases of D. hominis infestation in their animals, with cattle being the most affected host (100% of the affected farms). There was poor understanding of the D. hominis life cycle, vectors and control methods. A misuse of insecticides for the treatment of larval infestation by D. hominis was indicated.     

Time-dependent changes in plasma concentrations of 3-methylindole and blood concentrations of 3-methyleneindolenine-adduct in feedlot cattle

OBJECTIVE: To describe time-dependent changes in plasma concentrations of 3-methylindole (3MI) and blood concentrations of 3-methyleneindolenine (3MEIN)-adduct in feedlot cattle. ANIMALS: 64 yearling steers. PROCEDURES: Steers were assigned to 2 groups (32 steers/group). During the first 8 weeks, blood samples were collected from group 1 before the morning ration was fed, whereas samples from group 2 were collected 2 to 3 hours after the ration was fed. Blood samples were collected from all steers approximately 4 times/wk for 3 weeks and 3 times/wk for the subsequent 5 weeks. Samples were collected at the same time for all steers for an additional 10 weeks. Plasma samples were analyzed for 3MI concentrations. Blood samples collected from cattle in group 2 during the first 8 weeks were analyzed for 3MEIN-adduct concentrations. RESULTS: Mean blood concentration of 3MEIN-adduct increased to a maximum value on day 33 (0.80 U/microg of protein) and then decreased to a minimum on day 54 0.40 U/microg of protein). Plasma 3MI concentrations initially decreased and remained low until after day 54. Group-1 cattle had lower plasma 3MI concentrations, compared with concentrations for group-2 cattle. Blood 3MEIN-adduct concentrations and plasma 3MI concentrations were not associated with deleterious effects on weight gains. CONCLUSIONS AND CLINICAL RELEVANCE: Blood 3MEIN-adduct concentrations peaked during the period of greatest risk for development of bovine respiratory disease complex. Conversely, plasma 3MI concentrations decreased during the same period. Animal-to-animal variation in metabolic capacity to convert 3MI to 3MEIN may be of more importance than differences in plasma 3MI concentration.     

Short-term effect of therapeutic shoeing on severity of lameness in horses with chronic laminitis

OBJECTIVE: To evaluate the short-term effects of 4 therapeutic shoeing systems on lameness and voluntary limb-load distribution in horses with chronic laminitis. ANIMALS: 10 horses with chronic laminitis. PROCEDURES: A clinical trial was conducted that used a concurrent control, crossover design to evaluate the relative effectiveness of a standard flat shoe, fullered egg-bar shoe, heart-bar shoe, and modified equine digital support system to alleviate chronic lameness in horses. Therapeutic success was assessed during a 7-day period by use of subjective (Obel grade and clinical score) and objective (force-plate data) evaluations. RESULTS: Comparison of pretreatment and intertreatment control data indicated that disease status of the horses did not change during the course of the study. None of the therapeutic shoeing treatments used resulted in a significant change in severity of lameness. CONCLUSIONS AND CLINICAL RELEVANCE: Results were interpreted to imply that substantial clinical improvement should not be expected during the first 7 days after therapeutic shoeing for the specific shoes tested in this study. On the basis of our results, we hypothesize that when used as the lone indicator of therapeutic success, severity of lameness may not be a valid indicator.     

Use of low doses of ketamine administered by constant rate infusion as an adjunct for postoperative analgesia in dogs

OBJECTIVE: To compare indicators of postoperative pain and behavior in dogs with and without a low-dose ketamine infusion added to usual perioperative management. DESIGN: Prospective, randomized, blinded clinical study. ANIMALS: 27 dogs undergoing forelimb amputation. PROCEDURE: Dogs were anesthetized with glycopyrrolate, morphine, propofol, and isoflurane. Thirteen dogs were treated with ketamine IV, as follows: 0.5 mg/kg (0.23 mg/lb) as a bolus before surgery, 10 microg/kg/min (4.5 microg/lb/min) during surgery, and 2 microg/kg/min (0.9 microg/lb/min) for 18 hours after surgery. Fourteen dogs received the same volume of saline (0.9% NaCl) solution. All dogs received an infusion of fentanyl (1 to 5 microg/kg/h [0.45 to 2.27 pg/lb/h]) for the first 18 hours after surgery. Dogs were evaluated for signs of pain before surgery, at the time of extubation, and 1, 2, 3, 4, 12, and 18 hours after extubation. Owners evaluated their dogs' appetite, activity, and wound soreness on postoperative days 2, 3, and 4. RESULTS: Dogs that received ketamine infusions had significantly lower pain scores 12 and 18 hours after surgery and were significantly more active on postoperative day 3 than dogs that received saline solution infusions. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that perioperative administration of low doses of ketamine to dogs may augment analgesia and comfort in the postoperative surgical period.     

Sensitivity of test strategies used in the Voluntary Johne's Disease Herd Status Program for detection of Mycobacterium paratuberculosis infection in dairy cattle herds

OBJECTIVE: To evaluate sensitivities at the herd level of test strategies used in the Voluntary Johne's Disease Herd Status Program (VJDHSP) and alternative test strategies for detecting dairy cattle herds infected with Mycobacterium paratuberculosis. DESIGN: Nonrandom cross-sectional study. SAMPLE POPULATION: 64 dairy herds from Pennsylvania, Minnesota, Colorado, Ohio, and Wisconsin. Fifty-six herds had at least 1 cow shedding M. paratuberculosis in feces; the other 8 herds were free from paratuberculosis. PROCEDURE: For all adult cows in each herd, serum samples were tested for antibodies to M. paratuberculosis with an ELISA, and fecal samples were submitted for bacterial culture for M. paratuberculosis. Sensitivities at the herd level (probability of detecting infected herd) of various testing strategies were then evaluated. RESULTS: Sensitivity at the herd level of the testing strategy used in level 1 of the VJDHSP (use of the ELISA to test samples from 30 cows followed by confirmatory bacterial culture of feces from cows with positive ELISA result) ranged from 33 to 84% for infected herds, depending on percentage of cows in the herd with positive bacterial culture results. If follow-up bacterial culture was not used to confirm positive ELISA results, sensitivity ranged from 70 to 93%, but probability of identifying uninfected herds as infected was 89%. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that the testing strategy used in the VJDHSP will fail to identify as infected most dairy herds with a low prevalence of paratuberculosis. A higher percentage of infected herds was detected if follow-up bacterial culture was not used, but this test strategy was associated with a high probability of misclassifying uninfected herds.     

Occurrence of various immunoglobulin isotopes in horses with equine recurrent uveitis (ERU)

We investigated 30 healthy eyes and 41 eyes with ERU from 57 horses. The total immunoglobulin titers and titers of IgGa, IgGb, IgM were measured in aqueous humour, vitreous and serum using different ELISA techniques. Every sample investigated contained detectable amounts of immunoglobulins. Compared to control eyes significantly increased titers were found in the aqueous humour and vitreous of the ERU eyes for all immunoglobulin isotypes studied (p < or = 0.01). While IgM was detected in only 2 out of thirty aqueous humour and in none of the thirty vitreous samples of healthy eyes, 79.6% of samples of ERU eyes revealed considerable IgM titers. Changes of the IgGa/IgGb ratio in the eye as compared to that in the autologous serum was more frequent in affected than in healthy eyes. In contrast to the intraocular immunoglobulins there were no significant differences in immunoglobulin serum titers in healthy horses and those affected with ERU (p > 0.05). In conclusion, the results argue for a physiological appearance of immunoglobulins in the healthy eye. The increased titers of immunoglobulins in eyes stricken with ERU might be signs either of a local ocular production of antibodies and/or an increased permeability of intraocular barriers.     

Effect of the previously gravid uterine horn and postpartum interval on follicular diameter and conception rate in beef cows treated with estradiol benzoate and progesterone

An experiment was performed to evaluate the effect of the side of ovulation with respect to the previously gravid uterine horn on fertility of cows inseminated at one of two periods postpartum. All cows were treated with an intravaginal progesterone insert for 7 d and received estradiol benzoate (2 mg, i.m.) at the time of device insertion, prostaglandin F2alpha (25 mg, i.m.) at the time of device removal, and estradiol benzoate (1 mg, i.m.) 30 h after device removal. All cows were inseminated 28 to 30 h after the second treatment with estradiol benzoate, regardless of observed estrus. Cows treated in Period 1 received inserts at 16 to 20 d postpartum and were inseminated at 25 to 29 d postpartum. Cows treated in Period 2 received inserts at 26 to 30 d postpartum and were inseminated at 35 to 39 d postpartum. Diameter of the largest follicle at insert removal was greater in cows treated in Period 2 (10.1 +/- 0.3; mm +/- SEM) than in cows treated in Period 1 (9.1 +/- 0.3; P < .05). Diameter did not differ with the side of ovulation in respect to the previously gravid uterine horn. Diameter was greater in cows 5 to 9 (10.3 +/- 0.3) than in cows 3 to 4 (9.0 +/- 0.3) or 10 to 13 (9.4 +/- 0.6) yr of age (P < .01). The proportion of cows that ovulated from the ovary contralateral to the previously gravid uterine horn was greater (P < .05) than of those that ovulated from the ipsilateral ovary, and the incidence of ovulation was reduced in cows 3 to 4 yr of age (P < .01). Conception rate tended to be greater for ovulation from the ipsilateral compared with the contralateral ovary, relative to the previously gravid uterine horn (P < .10) and for ovulation from the right than the left ovary (P < .06). Conception rate was less if cows ovulated a follicle that was < 9 mm than a follicle > or = 9 mm in diameter at insert removal (P < .01) and was greater in cows inseminated in June than in April or May (P < .05). In conclusion, in cows in which estrus was synchronized at 25 to 39 d postpartum, ovulation from either the ovary ipsilateral to the previously gravid uterine horn, or the right ovary, tended to increase fertility.     

Expression and characterisation of equine interleukin 2 and interleukin 4

In the present study, we describe the expression of equine IL2 and IL4. The cDNA of equine IL2 or IL4 was cloned in a mammalian expression vector, containing c-terminal myc- and six histidines His(6)-epitopes for recognition and purification of equine cytokines. The vector constructs were used for transfection of chinese hamster ovary (CHO) cells. Purified equine cytokines were characterised by western blotting. Equine IL2 was secreted with a molecular weight of approximately 17.1kDa, whereas IL4 was expressed in three different sizes of 17.1, 19.6 and 22.1kDa, probably due to different glycosylation modifications. The biological activities of both cytokines were tested by proliferation assays using leukoagglutinin (LAG) prestimulated equine PBMC. Both, equine IL2 and IL4 induced dose-dependent lymphocyte proliferation. In contrast to IL4, IL2 supported the proliferation of B cells.