Laboratory diagnosis and disease occurrence in the current African swine fever eradication program in Spain, 1989–1991

The purpose of this study was to determine the rate of decline and geographical distribution by municipality of clinical and subclinical African swine fever (ASF) in the affected areas of Spain. A second aim was to evaluate the performance of diagnostic tests in the Spanish ASF eradication program. Clinical outbreaks were confirmed using both the direct and indirect immunofluorescence test (and if both were negative, by the hemabsorption test). The serological status of swine was determined by an enzyme-linked immunosorbent assay (ELISA) and suspect serum samples were confirmed by the immunoblot assay.

The number of clinical outbreaks (herds) of ASF for 1989, 1990 and 1991 was 170, 347 and 207, respectively. The numbers of municipalities within each affected province experiencing acute outbreaks for the same time periods were 49, 69 and 48, respectively. Serologically diagnosed animals positive for ASF were 1.1% of animals tested in 1989, 0.5% in 1990 and 0.8% in 1991. The corresponding positive predictive values of the standard ELISA test used were 99.0, 97.9 and 98.8, respectively. Similarly, the number of municipalities within each affected province experiencing serologically positive subclinically infected animals was 269, 178 and 147 for each of the years 1989, 1990 and 1991, respectively.     

Comparative histological study of the reproductive system of the female llama (Lama guanicoe glama). I. Ovary]

In the present study a cytological, histological and morphometrical comparison between the ovaries of the llama, the cow and the sheep is presented, at two phases of the ovarian cycle. There were found differences in the amount of primordial and primary follicles, the size of secondary follicles and follicular cells, and type and distribution of the connective tissue inside the stroma of the ovary. It would be necessary to study the fine structure of the ovary and the so-called "embryological remnants", for its permanent appearance in most (50%) of the ovaries.     

Effect of prostagladin F2 alpha on the bovine fetal ruminal wall in vitro

Strips of rumen wall from bovine fetuses were incubated in an organ bath with prostaglandin F₂ alpha (0.13 to 33.76 g/ml). The highest reactivity with a submaximal dose (17.03 g/ml) was observed in the period between 3.0 and 7.9 months of fetal age. A smaller response, but higher than in 1.0 to 2.9 months old fetuses, was observed in the 8.0 to 8.9 months fetuses. The period of the highest reactivity to prostaglandin F₂ alpha coincides with the age of onset of papillary morphogenesis and the period of highest reactivity to autonomic and putative transmitter drugs.     

Pharmacokinetics of carfentanil and naltrexone in domestic goats (Capra hircus).

Using a crossover study design, the pharmacokinetics of carfentanil and naltrexone after i.v., i.m., and s.c. administration were determined in eight domestic goats (Capra hircus). Serial blood samples were taken up to 120 hr after carfentanil administration, and the plasma drug concentrations were determined using liquid chromatography and mass spectroscopy. All goats were immobilized with 40 microg/kg carfentanil i.m., although the resulting neurologic effects varied considerably. Plasma profiles showed rapid carfentanil absorption and a simple biphasic decline for 12-48 hr. Naltrexone given at 100 mg naltrexone/mg carfentanil 30 min after carfentanil administration produced rapid reversal of immobilization after all routes of administration. Variable fluctuations in the naltrexone plasma concentrations during the first 2.5-3.5 hr were observed, followed by a more consistent biphasic decline. The time to standing was significantly shorter after i.v. compared with s.c. naltrexone, although the time difference (1 min) had little clinical relevance. No statistically significant differences between the naltrexone pharmacokinetic parameters measured for the three routes of naltrexone administration were identified, although the recoveries after i.m. administration were, subjectively, the smoothest. The carfentanil half-life did not differ significantly in the goats given naltrexone by different routes. Although it is currently recommended that the naltrexone dose be divided into s.c. and i.v. portions, this practice does not appear to offer any benefit.     

A filter-assisted culture method for isolation of Treponema spp. from bovine digital dermatitis and their identification by MALDI-TOF MS

Digital dermatitis (DD) is a major infectious foot disease of cattle worldwide. Some DD stages are associated with lameness, and the disease has significant economic and animal welfare consequences. The pathogenesis of the disease is not yet fully understood, but Treponema spp. have been associated consistently with clinical cases. Isolation of these fastidious bacteria is difficult and cumbersome. We describe an improved method enabling the culturing of the 3 Treponema spp. (T. pedis, T. phagedenis, and T. medium) from bovine foot specimens derived from DD lesions, using a combination of membrane filtering and subsequent growth on selective agar media. The entire procedure from sampling to verification of individual Treponema spp. takes up to 24 d. In addition, we established a MALDI-TOF MS–based identification method to be applied for confirmation of the different Treponema spp. This scheme provides an unambiguous, simple, and straightforward identification procedure for DD-associated Treponema spp.     

Effect of maternal cells transferred with colostrum on cellular responses to pathogen antigens in neonatal calves

OBJECTIVE: To assess the effect of maternal cells or cellular components on neonatal immune responses to intracellular pathogens in calves. ANIMALS: 15 Holstein calves. PROCEDURES: Calves were fed whole colostrum, frozen colostrum, or cell-free colostrum within 4 hours after birth. Leukocytes were obtained from calves before feeding colostrum and 1, 2, 7, 14, 21, and 28 days after ingestion. Proliferative responses against bovine viral diarrhea virus (BVDV) and mycobacterial purified protein derivatives were evaluated. Dams received a vaccine containing inactivated BVDV, but were not vaccinated against mycobacterial antigens. RESULTS: All calves had essentially no IgG in circulation at birth, but comparable and substantial concentrations by day 1. Calves that received whole colostrum had enhanced responses to BVDV antigen 1 and 2 days after ingestion of colostrum. In contrast, calves that received frozen colostrum or cell-free colostrum did not respond to BVDV. No differences were identified among the 3 groups in response to mycobacterial antigens. CONCLUSIONS AND CLINICAL RELEVANCE: Results indicated that transfer of live maternal cells from colostrum to neonatal calves enhanced responses to antigens against which the dams had previously responded (BVDV), but not to antigens to which the dams were na?ve (mycobacterial purified protein derivatives). Results suggested that cell-mediated immune transfer to neonates can be enhanced by maternal vaccination.     

In vitro effects of bethanechol on specimens of intestinal smooth muscle obtained from the duodenum and jejunum of healthy dairy cows

OBJECTIVE: To describe the in vitro effects of bethanechol on contractility of smooth muscle preparations from the small intestines of healthy cows and define the muscarinic receptor subtypes involved in mediating contraction. SAMPLE POPULATION: Tissue samples from the duodenum and jejunum collected immediately after slaughter of 40 healthy cows. PROCEDURES: Cumulative concentration-response curves were determined for the muscarinic receptor agonist bethanechol with or without prior incubation with subtype-specific receptor antagonists in an organ bath. Effects of bethanechol and antagonists and the influence of intestinal location on basal tone, maximal amplitude (A(max)), and area under the curve (AUC) were evaluated. RESULTS: Bethanechol induced a significant, concentration-dependent increase in all preparations and variables. The effect of bethanechol was more pronounced in jejunal than in duodenal samples and in circular than in longitudinal preparations. Significant inhibition of the effects of bethanechol was observed after prior incubation with muscarinic receptor subtype M(3) antagonists (more commonly for basal tone than for A(max) and AUC). The M(2) receptor antagonists partly inhibited the response to bethanechol, especially for basal tone. The M(3) receptor antagonists were generally more potent than the M(2) receptor antagonists. In a protection experiment, an M(3) receptor antagonist was less potent than when used in combination with an M(2) receptor antagonist. Receptor antagonists for M(1) and M(4) did not affect contractility variables. CONCLUSIONS AND CLINICAL RELEVANCE: Bethanechol acting on muscarinic receptor sub-types M(2) and M(3) may be of clinical use as a prokinetic drug for motility disorders of the duodenum and jejunum in dairy cows.     

Diagnostic and public health investigation of Mycobacterium tuberculosis infection in a dog in Ontario,Canada

A 4-y-old, female mixed-breed dog was presented to the Ontario Veterinary College for further evaluation of multiple pulmonary and hepatic masses, intrathoracic lymphadenitis, and recent development of a pyogranulomatous pleural effusion. Along with other comprehensive tests, a thoracic lymph node biopsy was performed, and Mycobacterium tuberculosis complex infection was confirmed by real-time PCR. The dog’s condition declined post-operatively, and euthanasia was elected. Postmortem examination confirmed severe granulomatous pneumonia, hepatitis, intrathoracic and intraabdominal lymphadenitis, omentitis, and nephritis. Line-probe assays performed on samples collected postmortem confirmed the species as M. tuberculosis. 24-loci MIRU-VNTR genotyping, spoligotyping, and whole-genome sequencing revealed relations to known human isolates, but no epidemiologic link to these cases was investigated. Given the concern for potential human exposure during this animal’s disease course, a public health investigation was initiated; 45 individuals were tested for M. tuberculosis exposure, and no subsequent human infections related to this animal were identified. Our case highlights the need for more readily available, minimally invasive testing for the diagnosis of canine mycobacteriosis, and highlights the ability of canid species to act as potential contributors to the epidemiology of M. tuberculosis infections.     

Extension of fresh-cut “Blanquilla” pear (Pyrus communis L.) shelf-life by 1-MCP treatment after harvest

‘Blanquilla’ pears processed as fresh-cut products are highly sensitive to browning and softening. Common postharvest methods, such as the use of antibrowning compounds and/or modified atmosphere packaging, fail to preserve ‘Blanquilla’ pear slices long enough to be marketable. However, treatment with 1-MCP before cutting and peeling considerably improved their textural properties (9.2 N vs. 1.1 N with and without 1-MCP treatment, respectively) and color (a* values of 1 vs. 5 after 15 d at 4 °C, for slices pear treated with 1-MCP and without treatment, respectively). These positive changes were closely related to a decrease in respiratory activity determined on whole pears after 3 months of storage in air at 0 ± 1 °C and 95% R.H. (0.40 ± 0.05 mmol CO₂ kg⁻¹ h⁻¹ vs. 0.77 ± 0.04 mmol CO₂ kg⁻¹ h⁻¹ with and without 1-MCP treatment, respectively) and ethylene production (1.18 ± 0.36 nmol C₂H₄ kg⁻¹ h⁻¹ vs. 5.751 ± 1.12 nmol C₂H₄ kg⁻¹ h⁻¹ for samples treated with and without 1-MCP, respectively). The use of 1-MCP allows fresh-cut ‘Blanquilla’ pears to be sold up to about 5 d after processing. Treatment with 1-MCP could be a viable alternative to common technologies for extending the shelf-life of ‘Blanquilla’ pears as a fresh-cut product.