Molecular definition of the bovine granulocytopathy syndrome: identification of deficiency of the Mac-1 (CD11b/CD18) glycoprotein

Leukocytosis (34,600 WBC/microliter of blood) was detected in an apparently healthy 7-day-old Holstein heifer. Analysis of blood samples obtained over the next 41 days revealed chronic progressive neutrophilia, which peaked at greater than 85% neutrophils and exceeded 100,000 WBC/microliter. In vitro assessment of isolated blood neutrophils obtained from the heifer at 38 and 45 days of age revealed selected functional abnormalities. Endocytosis of immunoglobulin-opsonized Staphylococcus aureus and killing of this test organism by the calf's neutrophils were significantly diminished, as were phagocytosis-associated superoxide generation, chemiluminescence activity, and myeloperoxidase-catalyzed iodination. Diminished H2O2 elaboration by the calf's neutrophils was evident during ingestion of opsonized zymosan or on exposure to phorbol myristate acetate. Extracellular release (secretion) of elastase during ingestion of zymosan was also diminished, although total cell content of elastase was normal, compared with that of neutrophils from age-matched calves, and granular or other morphologic abnormalities of the calf's neutrophils were not evident by ultrastructural examination. Abnormalities of random migration were inconsistently detected, and normal or high degree of antibody-dependent cytotoxicity or natural killing by the calf's neutrophils was observed. Similar in vitro assessment of neutrophils obtained from the calf's dam revealed no functional abnormalities. The calf died at 48 days of age, with persistent fever and chronic diarrhea, despite administration of antibiotics. Histologic examination at necropsy revealed large numbers of intravascular neutrophils in most tissues, including massive neutrophil sequestration in spleen. However, a striking lack of extravascular neutrophils was evident in inflamed submucosa adjacent to intestinal ulcers heavily contaminated with enteric microorganisms. Bone marrow examination revealed diffuse myeloid hyperplasia, but no other abnormalities.     

Ultrastructural studies of the lung of turkeys (Meleagris gallopavo) inoculated intratracheally with Escherichia coli.

To test the hypothesis that walls of air capillaries are a site for Escherichia coli to pass the air-blood barrier, fimbriated and nonfimbriated strains of E. coli were inoculated intratracheally into 18-day-old turkeys. Venous blood was cultured, and turkeys were necropsied from 0.5 to 8 hours post-inoculation. Lungs were processed for histopathology and electron microscopy. E. coli 078 was identified ultrastructurally using rabbit anti-lipopolysaccharide antibody and protein A-colloidal gold. All birds developed bacteremia; there was no significant difference between groups given fimbriated or nonfimbriated bacteria. Bacteria adhered to the plasma membrane of air capillary epithelial cells and were seen within vacuoles of portions of these cells that lined the fornices of air capillaries. Bacteria were also seen in the basement membrane at the basal surface of air capillary epithelial cells and, rarely, in vacuoles of subjacent endothelial cells. Infected granular and non-granular cells that lined air atria were necrotic 4 hours post-inoculation. Bacteria were within the overlying trilaminar substance and between reticular fibers of the interstitial stroma and pleura at 30 minutes post-infection and thereafter. Thus, the pulmonary air capillaries are a site for entrance of E. coli into the pulmonary blood capillaries, but fimbriae play little or no role in passage across the air-blood barrier.     

In-vitro studies on the break-down of chlormequat (chlorcholinchloride) in the rumen]

Under anaerobic conditions chlormequat did not undergo microbial breakdown in samples of rumen contents or rumen juice. Comparison with other forms of microbial breakdown of chlormequat, such as in soil, showed that oxidative processes were responsible.     

Virus in sheep's skin

In a double sense, the ovine gamma herpesvirus type 2 (OvHV-2) is a virus in sheep's skin. Not only is it present world wide in all sheep breeds but also it causes malignant catarrhal fever (MCF) in cattle pigs, elk, and bison. OvHV-2 cannot be propagated in cell culture. Therefore, new results from OvHV-2 research are based on molecular techniques and may be summarized as follows. OvHV-2 is transmitted by respiratory routes as well as by sexual intercourse. Lambs are infected within the first few months of life. Leucocytes, primarily latently infected lymphocytes, are responsible for disseminating the virus over the entire organism. On rare occasions, virus particles could be visualized by electron microscopy in explanted lymphocyte cultures. Structural antigens were detected by immunohistology in M-cells of diseased rabbits. Immunologically and cell biologically active genes have been detected on the viral genome.The products arising from those are thought to fine balance, the number of latently infected cells in sheep and to keep them alive without causing harm. Thus, it seems that this balance has been found through co-evolution, favoring both virus and natural host. In contrast, other host species that were exclude from the process of co-evolution, are bound to fall from MCF.     

Antimicrobial peptides and surfactant proteins in ruminant respiratory tract disease

In ruminants, respiratory disease is multifactorial and a leading cause of morbidity and mortality. Pulmonary innate immunity is the first line of defense for the respiratory tract. Alteration of regulation, expression, and function of these factors may be important to disease development and resolution. Many antimicrobial peptides and surfactant proteins are constitutively expressed in the respiratory tract and expression levels are regulated. Beta-defensins are cationic peptides with broad antimicrobial activity against bacterial, viral and fungal pathogens. Beta-defensins are primarily expressed in mucosal epithelia (and in some species leukocytes); where they may also participate in chemotaxis, wound repair and adaptive immune responses. Surfactant proteins A and D are secreted pulmonary surfactant proteins that have antimicrobial and immune regulatory activity. Anionic peptide is a constitutively expressed, aspartate-rich peptide that has antimicrobial activity and is most prominent during reparative epithelial hyperplasia. Regulation of these immune defense components by stress, pathogens, and inflammatory cytokines may play a role in the susceptibility to, severity and resolution of respiratory infection. The expression patterns of these molecules can be specific for host-species, class of pathogen and stage of infection. Understanding the regulation of antimicrobial peptide/protein expression will further enhance the potential for novel prophylactic and therapeutic modalities to minimize the impact of respiratory disease.     

Detection of novel papillomaviruses in canine mucosal, cutaneous and in situ squamous cell carcinomas

Papillomavirus (PV) DNA is frequently uncovered in samples of human skin squamous cell carcinomas (SCC). However, the role of these viruses in the development of such cancers in canine species remains controversial. While approximately 100 human PVs are known, only one single canine oral PV (COPV) has been identified and studied extensively. Therefore, we applied a narrow-range polymerase chain reaction (PCR) suitable for the detection of classical canine and feline PVs, as well as a broad-range PCR, which has been used for the detection of various novel PVs in humans, in order to analyse 42 paraffin-embedded samples, representing three different forms of canine SCCs. Ten samples of skin tissues with various non-neoplastic conditions served as controls. While none of the negative controls reacted positively, PV DNA was discovered in 21% of the tested SCC samples. Interestingly, the classical COPV was amplified from only one sample, while the other positive cases were associated with a variety of thus far unknown PVs. This study suggests that a fraction of canine SCC is infected with PVs and that a genetic variety of canine PVs exists. Therefore, these results will facilitate the future study of the role of PVs in the development of canine skin cancers.     

Evaluation of papillomaviruses associated with cyclosporine-induced hyperplastic verrucous lesions in dogs

OBJECTIVE: To determine whether cyclosporine A-induced hyperplastic skin lesions of dogs were associated with papillomavirus infections. ANIMALS: 9 dogs that were treated with cyclosporine A and developed hyperplastic skin lesions. PROCEDURE: History and clinical and histopathologic data were collected. Paraffin-embedded skin biopsy specimens from hyperplastic skin lesions were immunostained for common papillomavirus genus-specific structural antigens by use of a polyclonal rabbit anti-bovine papillomavirus type 1 antiserum. Sections from each tissue block underwent DNA extraction, and polymerase chain reaction (PCR) assays were performed with several sets of primers to amplify a wide range of papillomavirus DNA from humans and other animals. RESULTS: In 7 of 9 dogs, there were more than 10 hyperplastic skin lesions that microscopically resembled those of psoriasiform lichenoid dermatosis. In those dogs, results of testing for papillomavirus via immunohistochemical analyses and PCR assays were negative. In the other 2 dogs, there were only 1 and 3 verrucous lesions, and in those dogs, histologic evaluation revealed koilocytes and nuclear viral inclusions that were immunoreactive for papillomavirus antigens. Papillomavirus DNA was amplified from both dogs. One of the sequences was characteristic for the canine oral papillomavirus, whereas the other had similarities with the recently described canine papillomavirus 2. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, hyperplastic skin lesions occasionally develop during treatment with cyclosporine A. Most of the lesions resemble those of psoriasiform lichenoid dermatosis, although papillomavirus can be detected in some instances.     

The Anti‐Müllerian Hormone Profile is Linked with the In Vitro Embryo Production Capacity and Embryo Viability after Transfer but Cannot Predict Pregnancy Outcome

The current study investigated the possibility of using the AMH concentration as a predictor of the ability of Korean Hanwoo cows to produce cumulus‐oocyte complexes, embryos that survive after transfer as well as the pregnancy outcome of surrogates. Eight sessions of ovum pick‐up (OPU) were performed with 19 donor cows at an interval of 3–4 days. Antral follicle count (AFC), oocyte quality and in vitro embryo development were recorded for each cow. Embryos produced from cows with different AMH profiles were transferred into recipients (n = 96). Cows in the high (≥0.25 ng/ml) and intermediate (0.1≥ to <0.25 ng/ml) AMH groups had a significantly higher AFC per OPU session (20.40 ± 1.36 and 16.91 ± 1.52, respectively; mean ± standard deviation) than cows in the low AMH group (<0.1 ng/ml; 12.19 ± 2.14). In addition, more cumulus‐oocyte complexes per donor were recovered in the high (11.46 ± 1.22) and intermediate (7.38 ± 0.83) AMH groups than in the low AMH group (4.77 ± 0.44). The percentage of oocytes reached blastocyst stage was significantly higher in the intermediate (47.0%) and high (38.5%) AMH groups than in the low AMH group (32.3%). The number of embryos produced per cow was higher in the high (3.9 ± 0.2) and intermediate (6.9 ± 0.6) AMH groups than in the low AMH group (2.2 ± 0.3). The percentage of embryos that gave birth to viable calves when transferred into recipients was higher for those derived from cows in the intermediate AMH group (50.7%) than for those derived from cows in the low (35.7%) and high (36.4%) AMH groups. In conclusion, a single measurement of AMH concentration predicted the in vitro embryo production potential of donor Korean native cows before OPU and is linked with embryo viability after transfer into recipients.     

Die Stickstoffdüngung noch besser auf das von Jahr zu Jahr Schwankende natürliche N-Angebot aus dem N-Pool des Bodens abstimmen

Es wird eine Methode dargestellt, die auf der Grundlage der Höhe der Wärmesumme eine differenzierte Möglichkeit der N-Düngung ableitet. In Abhängigkeit von der Wärmesumme und der Bodenfeuchte ergibt sich eine differenzierte N-Düngerbereitstellung aus dem Boden. Diese N-Mineralisation aus dem Bodenstickstoff ist bei der Bemessung einer zweiten Gabe zu berücksichtigen. Die ermittelte N_min-Menge entscheidet über die Notwendigkeit einer Nachdüngung im Sechs- bis Achtblattstadium des Maises. Für ein hohes Ertragspotential von 120 dt/ha Körnermais (86% TS) sollte ein N_min-Wert um 220 kg bei einer Wärmesumme von 310°C ausgewiesen werden. Bei diesen Zahlen ist keine Nachdüngung erforderlich.