Immunohistochemical study of the inflammatory infiltrate associated with feline cutaneous squamous cell carcinomas and precancerous lesions (actinic keratosis).

The distribution of T lymphocytes (CD3+), B lymphocytes (CD79+), immunoglobulin-containing plasma cells (IgG, IgM and IgA), macrophages (Mac387+) and MHC Class II antigen was analysed in the inflammatory infiltrate associated with cutaneous squamous cell carcinomas (SCC) from 23 cats. Peri-tumoural skin (12 cases) and precancerous lesions of actinic keratosis (nine cases) were also evaluated for the expression of MHC Class II. The results revealed that an abundant inflammatory infiltrate was associated with the majority of SCC. This infiltrate was composed mainly of CD3+ T lymphocytes, B cells (CD79+) and IgG-bearing plasma cells, and the intensity of infiltration increased with the degree of invasiveness of the tumour. The number of CD3+ T cells and CD79+ cells was significantly increased in well-differentiated SCC compared with moderately differentiated tumours, whereas the number of IgM+, IgA+ plasma cells and Mac387+ macrophages was low or moderate and did not change significantly with histologic grade or invasiveness. MHC Class II antigen was expressed by infiltrating lymphocytes and macrophages, and by fibroblasts. A variable number of neoplastic cells (10% to 80%) in 10 SCC, and keratinocytes of basal layers in seven of nine cases of actinic keratosis also expressed MHC Class II, whereas keratinocytes of normal skin were always negative for this antigen. These results suggest that CD3+ T lymphocytes, CD79+ B cells and IgG-bearing plasma cells may participate in down-regulation of tumour growth, since these cell types were particularly numerous in well-differentiated and mildly invasive SCC, as well as in actinic keratosis. The expression of MHC Class II by neoplastic cells could enhance this local anti-tumour immune response.     

Characterization of the cDNA and genomic DNA sequence encoding for the platelet integrin alpha IIB and beta III in a horse with Glanzmann thrombasthenia

Glanzmann thrombasthenia (GT) is characterized by a defect of platelet aggregation. This autosomal recessive genetic disorder is caused by an abnormality of the platelet glycoprotein receptors alpha IIb or beta III. Recently, we identified a horse with clinical and pathological features of GT. The aim of this study was to describe this case of GT at the molecular level. A point mutation from G to C in exon 2 of ITGA2B causing a substitution of the expected amino acid arginine 72 (Arg(72)) by a proline (Pro(72)) was encountered. This amino acid change may result in abnormal structural conformations that yield an inactive alpha IIb subunit. The genomic DNA analysis showed that this horse was homozygous for the missense mutation.     

Campylobacter from sows in farrow-to-finish pig farms: risk indicators and genetic diversity

Sows have been identified as a source of Campylobacter contamination in piglets. We carried out a one-year study, in 2008, at 53 farrow-to-finish farms in Brittany, France, to determine the proportion of sows excreting Campylobacter. We also determined the genotypes of the Campylobacter isolates. Moreover, Generalized Estimating Equations including repeated effects were used to assess the association between management practices and farm characteristics, and risk of Campylobacter shedding by sows. Per farm, 10 feces samples from sows were collected from selected sites (maternity, service area, gestation area) on the farms. Campylobacter isolates were identified by PCR and typed by PFGE. Campylobacter was detected in 25.1% of the 530 samples from sows, and 67% of the 53 pig farms had at least one positive sample (of 10 taken). All the Campylobacter isolates belonged to the Campylobacter coli species. They displayed a very high level of genetic diversity, also inside farms and few genotypes were common to several farms. Warmer months, large farms, and individual housing for sows were identified as risk indicators of Campylobacter shedding by sows. A short delay between sampling and treatment of the samples should be considered, to improve the detection of the bacterium in the feces samples.     

Additivity of effects from dietary copper and zinc on growth performance and fecal microbiota of pigs after weaning

Four experiments were conducted to determine the interactive effects of pharmacological amounts of Zn from ZnO and Cu from organic (Cu-AA complex; Cu-AA) or inorganic (CuSO(4)) sources on growth performance of weanling pigs. The Cu was fed for 4 (Exp. 1) or 6 (Exp. 2, 3, and 4) wk after weaning, and Zn was fed for 4 (Exp. 1) or 2 (Exp. 2, 3, and 4) wk after weaning. Treatments were replicated with 7 pens of 5 or 6 pigs per pen (19.0 ± 1.4 d of age and 5.8 ± 0.4 kg of BW, Exp. 1), 12 pens of 21 pigs per pen (about 21 d of age and 5.3 kg of BW, Exp. 2), 5 pens of 4 pigs per pen (20.3 ± 0.5 d of age and 7.0 ± 0.5 kg of BW, Exp. 3), and 16 pens of 21 pigs per pen (about 21 d of age and 5.7 kg of BW, Exp. 4). In Exp. 1 and 2, Cu-AA (0 vs. 100 mg/kg of Cu) and ZnO (0 vs. 3,000 mg/kg of Zn) were used in a 2 × 2 factorial arrangement. Only Exp. 1 used in-feed antibiotic (165 mg of oxytetracycline and 116 mg of neomycin per kilogram feed), and Exp. 2 was conducted at a commercial farm. In Exp. 3, sources of Cu (none; CuSO(4) at 250 mg/kg of Cu; and Cu-AA at 100 mg/kg of Cu) and ZnO (0 vs. 3,000 mg/kg of Zn) were used in a 3 × 2 factorial arrangement. In Exp. 4, treatments were no additional Cu, CuSO(4) at 315 mg/kg of Cu, or Cu-AA at 100 mg/kg of Cu to a diet supplemented with 3,000 mg/kg of Zn from ZnO and in-feed antibiotic (55 mg of carbadox per kilogram of feed). In Exp. 1 and 2, both Zn and Cu-AA improved (P < 0.001 to P = 0.03) ADG and ADFI. No interactions were observed, except in wk 1 of Exp. 2, where Zn increased the G:F only in the absence of Cu-AA (Cu-AA × Zn, P = 0.04). A naturally occurring colibacillosis diarrhea outbreak occurred during this experiment. The ZnO addition reduced (P < 0.001) the number of pigs removed and pig-days on antibiotic therapy. In Exp 3, ADFI in wk 2 was improved by Zn and Cu (P < 0.001 and P = 0.09, respectively) with no interactions. In wk 1, G:F was reduced by ZnO only in the absence of Cu (Cu × Zn, P = 0.03). Feeding Zn decreased fecal microbiota diversity in the presence of CuSO(4) but increased it in the presence of Cu-AA (Cu source × Zn, P = 0.06). In Exp. 4, Cu supplementation improved the overall ADG (P = 0.002) and G:F (P < 0.001). The CuSO(4) effect on G:F was greater (P < 0.001) than the Cu-AA effect. Our results indicate that pharmacological amounts of ZnO and Cu (Cu-AA or CuSO(4)) are additive in promoting growth of pigs after weaning.     

Post-mortem examination and laboratory-based analysis for the diagnosis of bovine tuberculosis among dairy cattle in Ecuador

Veterinary inspection in slaughterhouses allows for the detection of macroscopic lesions reminiscent of bovine tuberculosis, but the presence of Mycobacterium bovis must be confirmed by laboratory methods. This study aimed at comparing the performances of the standard diagnostic tools used to identify M. bovis in tissue specimens sampled from suspicious animals. During a two years period, 1390 cattle were inspected at the Machachi abattoir in the Mejia canton - Ecuador. A total of 33 animals with granulomatous lesions were detected, representing 2.33% (16/687) and 2.42% (17/703) animals examined in 2007 and 2008, respectively. Ninety-four tissue specimens were sampled and screened for the presence of mycobacteria. Acid-fast bacilli were identified in one third of the suspicious cattle (11/33) and suggestive microscopic lesions in 27.3% (9/33) of the samples examined by direct microscopy and histopathology, respectively. Culturing on Stonebrink medium and 16S-rRNA-based polymerase chain reaction (PCR) yielded 36.4% (12/33) and 27.3% (9/33) of positives, respectively. Compared to culture, other diagnostic procedures displayed a lower sensitivity, with 56.5% for PCR, and 43.5% for direct microscopy and histopathology; however, the specificity was higher (94.4% for PCR and microscopy, and 97.2% for histopathology). We conclude that reliable post-mortem laboratory testing either requires the combination of a set of available diagnostic tools or necessitates the development of improved new-generation tools with better sensitivity and specificity characteristics.     

Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis.

Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR analysis and histologic examination suggested a leptospiral infection in the newborn foal. At the moment of death, the infection coexisted with a streptococcal-associated aspiration bronchopneumonia and postpartum septicemia. These findings indicate that the PCR assay based on the amplification of the hap1 gene represents a useful tool for specific detection of pathogenic leptospira in field samples taken from horses.     

Interindividual variability in plasma concentrations after systemic exposure of swine to dietary doxycycline supplied with and without paracetamol: a population pharmacokinetic approach

Anorexigenic substances released during infection may hinder the therapeutic efficacy of in-feed antibiotics. Paracetamol (acetaminophen; PARA) inhibits the anorexia of infection and seems to improve the clinical efficacy of doxycycline (DOX) against bacterial respiratory disease in swine herds. In order to verify whether PARA selectively stimulates intake of DOX-medicated feed in diseased pigs, we documented the pharmacokinetics (PK) of DOX when coadministered with PARA and examined the effect of in-feed PARA on the interindividual variability in plasma concentrations after systemic exposure to in-feed DOX in swine herds with respiratory disease. Systemic exposure to DOX was measured with the area under the curve (AUC) of its plasma concentrations over time. First, a rich-sampling PK study of in-feed and i.v. DOX (10 mg/kg of BW) and PARA (30 and 10 mg/kg of BW, respectively) was performed on 5 pigs. The PK profiles of in-feed DOX were used in mathematical simulations to determine 5 optimal sampling times for the farm-based population PK study. A randomized, blind, parallel PK study was performed in 2 herds with bacterial respiratory disease, where liquid feed was fortified with DOX alone (5 mg x kg of BW(-1) x meal(-1)) or combined with PARA (15 mg x kg of BW(-1) x meal(-1)). Medicated meals were given twice, 12 h apart, to group-housed growing pigs (n > 50 pigs x treatment(-1) x herd(-1), totaling 215 pigs). Plasma concentrations of DOX and PARA were measured with HPLC. At variance with our expectations, PARA decreased (P = 0.069) mean AUC of in-feed DOX and did not decrease its variability (P > 0.34). Mean AUC of DOX increased with feed intake and with initial exposure to DOX, and was greater in sick animals. Therefore, symptomatic PARA-induced improvement in bacterial respiratory disease control with DOX is more likely caused by its analgesic/antipyretic effects than by its orexigenic effect. Interindividual variation in the AUC of DOX was large in pigs given group medication, even when sufficient feeding space was allowed and the amount of feed offered was greater than their requirements. Therefore, future studies to improve the efficacy of group antibiotic therapy should focus on feeding behavior characteristics as well as biopharmaceutical properties of medicated feeds.     

Pathological findings in a captive colony of maras (Dolichotis patagonum)

This paper describes the causes of death of 54 maras (Dolichotis patagonum) in a captive colony in Mexico over a period of seven years. There were 35 adults, 11 juveniles, five neonates, two fetuses and one stillbirth--27 males, 21 females and six whose sex was not determined. Trauma was the cause of 25 deaths, and there were eight cases of fatal bacterial infection. Besnoitiosis was the only parasitic disease found frequently (13 cases), and was associated with fatal interstitial pneumonia in three juveniles. Right-sided hypertrophic cardiomyopathy attributed to high altitude was observed in 26 maras, and in three cases death was attributed to acute cardiac dysfunction. Two maras died of disseminated histoplasmosis and two of hyperthermia. Additional causes of death included one case each of uterine torsion, intestinal intussusception, aspiration pneumonia and hydranencephaly. Gastric erosions with luminal haemorrhage were found in 27 of the maras and splenic lymphoid depletion in 20, changes that were attributed to stress.