Immune dysregulation in atopic dermatitis

Atopic dermatitis (AD) is a chronic inflammatory skin disease in humans and dogs with comparable clinical features. Comparative studies of immunological events in the pathogenesis of AD may contribute to understanding of the disease in dogs and to development and evaluation of immunomodulatory strategies of relevance to both species.Both allergen-specific as well as non-specific mechanisms contribute to the disease development. AD skin lesions are proposed to be initiated by activation of allergen-specific Th2-type cells, potentially influenced by local cutaneous factors. In the chronic stage of skin lesions reactivity may change into a Th1-type, e.g. driven by eosinophil derived IL-12. Analyses of these processes in course of time were performed in both spontaneous as well as in experimentally induced lesions (i.e. atopy patch test (APT) lesions). In the present paper, the immunological events as reported for human and canine AD are summarized and compared.     

Development of a PCR test for the identification of Staphylococcus intermedius based on the 16S rDNA sequence

The development of a PCR assay based on the 16S ribosomal RNA gene (rDNA) sequence was carried out for the identification of Staphylococcus intermedius. Sixty-six strains of S. intermedius, 70 of Staphylococcus aureus and 2 of Staphylococcus hyicus were examined for the assay. The 16S rDNA, of which the PCR target fragment makes up 901 bp corresponding to the sequence data of the gene, was detected in all strains of S. intermedius, but it was not detected in any strains of either S. aureus or S. hyicus. These results suggest that the PCR allows a simple and precise identification of S. intermedius.     

Evaluation of 65% permethrin spot-on for prevention of canine visceral leishmaniasis: effect on disease prevalence and the vectors (Diptera: Psychodidae) in a hyperendemic area

A study was designed to examine the effect of 65% permethrin spot-on on the prevalence of canine visceral leishmaniasis and the abundance of sand flies in two neighborhoods in Corumbá, Mato Grosso do Sul, Brazil known to have a high prevalence of visceral leishmaniasis. An enrollment survey was conducted to determine the prevalence of visceral leishmaniasis. Area residents were provided with information about the project; the study area was defined, and all dogs (160 in Cristo Redentor and 230 in Popular Velha) identified in the study area were enrolled. Three 65% permethrin spot-on treatments (June 15-25, July 13-15, and August 10-12) were administered to 150, 110, and 99 dogs, respectively, in Popular Velha, according to label recommendations. Dogs in Cristo Redentor were untreated controls. Visceral leishmaniasis was diagnosed periodically by indirect immunofluorescence assay. A reduction in canine visceral leishmaniasis prevalence was observed at the Popular Velha site. The infection rate for treated dogs 1 month following the final treatment was approximately 50% reduced from that observed before treatment(19.3% vs 9.6%). Conversely, the infection rate at the control site was more than 80% higher at the September sampling than that observed pretreatment (4.1% vs 7.4%). Similar numbers of sand flies were captured and identified from both sites throughout the study. The results suggest that regular use of 65% permethrin during months of high risk for canine visceral leishmaniasis can be a useful strategy for reducing the prevalence of this disease in hyperendemic areas. It should be stressed, however, that the success of this strategy depends not only on the efficacy of the product itself but also on the adoption of other control measures and on economic variables, considering the low purchasing power of the populations living in higher-risk neighborhoods.     

Bilateral neuroepithelial choristomas of the optic disc in a cynomolgus monkey (Macaca fascicularis): a case report

A 6-year-old intact male cynomolgus monkey of Chinese origin was received at the Sierra Biomedical Facility. While physical examination revealed good body condition with no abnormalities, routine ophthalmic examination revealed bilateral proliferative optic neuropathy involving the dorsal aspect of the optic disc. No changes were noted in the appearance of the lesions over 8 months, and fluoroescein angiography revealed no abnormalities other than obstruction of the view of the retinal vessels by the lesions. Histopathologic studies revealed characteristics consistent with a diagnosis of bilateral neuroepithelial choristoma.     

Evaluation of L-selectin expression and assessment of protein tyrosine phosphorylation in bovine polymorphonuclear neutrophil leukocytes around parturition

Impaired polymorphonuclear neutrophil leukocyte (PMN) function around parturition has been associated with increased clinical mastitis in dairy cows. Rolling and attachment of PMN to the endothelium is the first step in the recruitment process and is accomplished by interaction between L-selectin on PMN and its ligand on endothelial cells. Furthermore, tyrosine phosphorylation is involved in the initiation of many PMN functions. The objective of this work was to determine changes in expression of L-selectin and tyrosine phosphorylation in the perinatal period. Eight clinically healthy Holstein cows were used as PMN donors at d-21, -14, -7,0 (calving), +1, +2, +7, +14, +28. Evaluation of L-selectin expression was carried out on activated and resting PMN. Anti-bovine L-selectin monoclonal antibody (MAB) and flow cytometric analysis were used to measure the percentage of PMN fluorescing and receptor expression (log mean fluorescent channel, LMFC). Activated and resting PMN showed similar trends in % PMN fluorescence and LM FC. The percentage of PMN fluorescing tended to decrease at parturition, followed by a significant increase at d +14 and +28 (P < 0.02). For LMFC a decrease was observed on d +1 followed by an increase through d +28 (P < 0.01). Protein tyrosine phosphorylation of lysates prepared from PMN isolated throughout the study was detected by electrophoresis and western blotting using anti-phosphotyrosine MAB. Several protein bands were tyrosine phosphorylated. Two of these bands (42-44 kDa and 90 kDa) varied in intensity over time. The intensity of the 42-44 kDa band gradually increased from d -7, peaked at d +7 (P < 0.03), and steadily decreased to d +28 (P < 0.02). Antibody to activated mitogen protein kinase reacted with the 42-44 kDa band. Reduced PMN function during the periparturient period could be related to reduced L-selectin adhesion molecules on the cell surface, and to modulation in the phosphorylation of functionally important molecules.     

Ultrasonographic evaluation of urinary bladder in normal,fern fed and enzootic bovine haematuria-affected cattle

A total of 19 adult hill cattle of both sexes were subjected to trans-rectal ultrasound scanning of urinary bladder to evaluate bladder wall thickness and the presence of space-occupying lesions. The animals were divided into four groups. Eight apparently healthy hill cattle maintained under standard ration served as control (group I) and the remaining II animals were divided into three groups (II, III and IV). Group II animals (n = 8) were fed with different type of ferns which were further divided into subgroups II-P, -D and -B and fed with Polystichum squarrosom (n = 2). Dryopteris juxtaposita (n = 2) and Pteridium aquilinum (n = 4) ferns, respectively. The one animal in group III was a natural case of enzootic bovine haematuria (EBH) and the two animals in group IV were natural cases of microscopic EBH fed with Polystichum squarrosum fern. In group I animals, the average bladder wall thickness was 1.45 mm. The delineation of the bladder wall was uniformly smooth and the echo pattern of the bladder was homogeneously black, which was suggestive of clear urine content. In group II (P, D and B) the average bladder wall thickness of the six animals was 1.87 mm and the sonographic features were within normal limit when compared with controls. In two of the animals of group II-B, the bladder wall was apparently thick (4.36 mm) and there was no intraluminal mass except at one or two focal elevated points. Animals of groups III and IV showed the average bladder wall thickness of 4.86 mm and were characterized by the presence of irregular sessile masses extending into the bladder lumen. The homogeneous anechoic area was reduced centrally due to the presence of a hypoechoic soft tissue mass all around the bladder wall. Post-sonographic urinalysis, biopsy and necropsy of selected cases further confirmed the sonographic findings.     

The influence of age and gender on haematological parameters in Lipizzan horses

Haematological parameters [red blood cell count (RBC), white blood cell count (WBC), packed cell volume (PCV), haemoglobin concentration, mean cell volume (MCV), mean cell haemoglobin (MCH) and mean cell haemoglobin concentration (MCHC)] in resting Lipizzan horses were determined for 143 stallions, 104 mares and 25 foals. The mean RBC and WBC values in Lipizzans were in the lower part of the normal range for warm-blooded horses. The mean PCV, MCV and MCH values were higher, but the mean haemoglobin concentration and MCHC values were lower than reported for other warm-blooded horses. In foals, the mean RBC, WBC, PCV, haemoglobin concentration and MCHC values were higher, whereas MCV and MCH were lower than in older animals. Results indicating a significant decrease in WBC (P < 0.01) and an increase in MCV, MCH and MCHC (P < 0.05) with increasing age are consistent with some other reports on warm-blooded horses. The age-related variations in RBC and PCV were less marked. Contrary to some reports, RBC (P < 0.01), WBC and haemoglobin concentration (P < 0.001) were higher in Lipizzan stallions than in mares, but differences in MCHC, MCH and MCV were insignificant. The specific haematological values determined in Lipizzans are presumably a result of selection and should be taken into consideration when dealing with this race of horses.     

Susceptibility of trypanotolerant West African Dwarf goats and F1 crosses with the susceptible Sahelian breed to experimental Trypanosoma congolense infection and interactions with helminth infections and different levels of diet

Forty pure West African Dwarf (WAD) goats and 35 of its F1 crosses with the Sahelian breed were used in a multifactorial experimental design to evaluate the effects of an experimental Trypanosoma congolense infection and interactions with natural helminth infections and different levels of diet on health and productivity of these two breeds. Trypanosome infection caused a severe drop in packed cell volume (PCV), but this was not significantly affected by breed. Neither deworming nor diet had any effect on the course of anaemia after trypanosome infection. The mean score of parasitaemia tended to be higher in crossbreeds than in WAD goats although this was not significant (P>0.05). Similarly, the antibody response to trypanosome infection was not significantly different between breeds. Parasitaemia level was significantly influenced by the level of diet with the group under high supplementation having a higher mean parasitaemia score than the group under low supplementation. Weight loss due to trypanosome infection tended to be greater in crossbreeds than in WAD goats (P>0.05).During this study, there was no difference in mean helminth egg output between crossbred and WAD goats. However, between weeks 4 and 10 after trypanosome infection (corresponding to a period of heavy rainfall and highly infective pastures), the mean egg output was higher in the crossbreeds.The immunosuppressive effect of trypanosome infections was revealed by a lower antibody response to Haemonchus contortus in infected animals compared to the uninfected controls. Trypanosome infection tended to increase strongyle egg output. This study did no reveal any superior trypanotolerance of WAD goats compared to crossbreeds.     

A cross-sectional coprological survey of liver flukes in cattle and sheep from an area of the southern Italian Apennines

A cross-sectional coprological survey of liver flukes (Fasciola hepatica and Dicrocoelium dendriticum) was conducted on 81 bovine farms and 197 ovine farms with animals pasturing in an area (3971 km(2)) of the southern Italian Apennines. The farms were selected to be uniformly distributed throughout the study area using geographical information system (GIS) software. Between June 1999 and March 2000, faecal samples were collected from 975 cattle and 3940 sheep and examined using a modified McMaster technique. The results were subjected to statistical analysis and point distribution maps (PDMs) were drawn by GIS.Cattle of 9 of the 81 (11.1%) farms were positive for F. hepatica and of 43 (53.1%) for D. dendriticum. Sheep of 8 of the 197 (4.1%) farms were positive for F. hepatica and of 133 (67.5%) for D. dendriticum. Co-infection was found in cattle of 2 (2.5%) farms, and in sheep of 8 (4.1%) farms.The findings of the present survey show that D. dendriticum was the predominant liver fluke found in cattle and sheep with respect to egg count numbers for both farms and animals. In addition, the general trends of the PDMs show that D. dendriticum was widely and homogeneously spread throughout the study area, whereas F. hepatica was present only in a few concentrated zones of the study area that had both positive bovine and positive ovine farms.     

Prevalence of and resistance to anti-microbial drugs in selected microbial species isolated from bulk milk samples

The prevalence of strains of Staphylococcus aureus, coagulase-negative (CN) staphylococci, Listeria monocytogenes, Escherichia coli, Enterococcus faecalis, E. faecium and Bacillus cereus, was investigated in 111 bulk milk samples. Staphylococcus aureus was isolated from 38 samples, CN staphylococci from 63 samples, E. coli from 49 samples, E. faecalis or E. faecium from 107 samples, and L. monocytogenes from two samples. Bacillus cereus was not found in any of the samples and three samples were free of any of the selected species. Sensitivity to the anti-microbial drugs amikacin, ampicillin, ampicillin + sulbactam, cephalothin (CLT), cephotaxime, clindamycin, chloramphenicol (CMP), co-trimoxazole, erythromycin (ERY), gentamicin, neomycin, norfloxacin, oxacillin, penicillin, streptomycin (STR), tetracycline (TTC) and vancomycin was tested using the standard dilution technique. Minimum inhibitory concentration (MIC) characteristics (MIC50, MIC90, MIC range) were determined for each microbial species. Resistance against one or more anti-microbial drugs was found in 93% of S. aureus, 40% of CN staphylococci, 73% of E. coli, 88% of E.faecalis, 55% of E.faecium, and one L. monocytogenes strain. Most of the strains, particularly enterococci, were resistant to STR, TTC, and ERY (MIC50 4 microg/ml). A high percentage of staphylococci were resistant to beta-lactam antibiotics. High resistance to CLT was found in 11 strains of E. coli (MIC 256 microg/ml) and strains resistant to CMP (MIC90 16 microg/ml) were detected. The highest numbers of resistance phenotypes were found in E. coil (16) and CN staphylococci (12). Eighteen identical resistance phenotypes were demonstrated in indicator bacteria (E. coli, E. faecalis, E. faecium) and pathogens (S. aureus, CN staphylococci) isolated from the same bulk milk sample. The obtained resistance data were matched against the herd owners' information on therapeutic use of the drugs. This confrontation could not explain the findings of strains resistant to ERY or CMP. Our findings are evidence of selection of resistant strains among not only pathogenic agents, but also among indicator bacteria which can become significant carriers of transmissible resistance genes.