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Biomedical subjects

G Leitner

Publications and source records attributed to G Leitner.

At least 19 recordsLinked to original sources

Genetic analysis of persistency in the israeli holstein population by the multitrait animal model.

Persistency was defined as the predicted milk production 180 d after peak divided by peak production (in %). Heritability of persistency in a multitrait analysis including parities 1 through 5 increased from 0.16 to 0.27 from first through third parity, and then declined through fifth parity. Genetic correlations for persistency between consecutive parities were all > 0.8. First-parity genetic correlations of the traits included in the Israeli selection index with persistency were all < 0.1, except for fertility and herdlife, which were 0.20 and 0.25; whereas second-parity genetic correlations of persistency with the 3 milk production traits were all > 0.34, and the genetic correlation with fertility was only 0.10. The genetic correlation between second-parity persistency and herdlife was 0.58. Persistency in the Israeli Holstein population was analyzed by the multitrait animal model. The genetic trend since 1985 for the multiparity index was 0.22% persistency/yr, even though there was no direct selection on persistency.

Animals↗

Changes in milk composition as affected by subclinical mastitis in sheep.

The mechanism of the effects of glandular-level subclinical mastitis in dairy sheep on milk yield and on its composition as expressed in curd yield was studied. Thirty-six Israeli-Assaf dairy sheep with one udder half infected with identified coagulase-negative staphylococci and the contralateral gland free of bacteria were chosen. The milk yield of the infected halves was significantly lower than that of the uninfected ones (0.36 vs. 0.76 kg/milking). The somatic cell count and N-acetyl-beta-D-glucosaminidase activity were significantly higher in the infected halves than in the uninfected ones. The plasminogen activator and plasmin (PL) activities were significantly higher in the infected glands than in the uninfected ones, whereas plasminogen (PLG) activity and the ratio PLG:PL were significantly lower in the infected glands. Concentrations of Ca2+ did not differ, whereas Ca2+ activity was significantly lower and proteose peptone concentration was 2.4 times as high in the infected glands than in the uninfected ones. Curd yield was significantly lower in the infected glands than in the uninfected ones.

Animals↗

Changes in milk composition as affected by subclinical mastitis in goats.

The mechanism of the effects of subclinical mastitis (SM) at the glandular level in dairy goats on milk yield and its composition as expressed in curd yield (Yc) was studied. Twenty-five Israeli goats of various crossbreeds were chosen; one udder half was naturally infected with identified coagulase-negative staphylococci, and the contralateral gland was free of bacteria. The milk yield of the infected halves was significantly lower than that of the uninfected ones. Somatic cell count and N-acetyl-beta-D-glucosaminidase activity were significantly higher in the infected halves. The lactose concentration in the infected glands was significantly lower than that in the uninfected ones, casein concentrations did not differ, and the whey protein and albumin concentrations were significantly higher in the infected glands. Plasmin activity was significantly higher in the infected glands, whereas plasminogen activity was undetectable. Concentrations of Ca2+ did not differ, whereas Ca2+ activity was significantly lower in the infected glands. The proteose peptone concentration was 1.5 times as great in the infected glands as in the uninfected ones. The Yc was significantly lower in the infected halves, and clotting time was significantly longer. The mechanisms of the effects of SM on milk yield and Yc in goats and sheep are discussed and compared.

Acetylglucosaminidase↗

Staphylococcus aureus vaccine against mastitis in dairy cows, composition and evaluation of its immunogenicity in a mouse model.

Bovine mastitis caused by Staphylococcus aureus (S. aureus) is a most important infection disease that affects both the quality and the quantity of milk production. Antibiotic therapies formulated for intramammary use are generally unsuccessful in eliminating existing S. aureus infections. Vaccination is a logical approach to the control of S. aureus udder infections. However, to date commercially available S. aureus vaccine have shown limited efficacy under field conditions, mainly due to the paucity of information regarding relevant antigens which will induce a broad spectrum immunization. In the present paper the attempt to develop a new vaccine designated MASTIVAC I is described. MASTIVAC I is composed of three strains of S. aureus namely: VLVL8407; ZO3984 and BS449 which were isolated from clinical and sub-clinical cases of bovine mastitis. A mouse model was used to evaluate the S. aureus specific antibody production and protection of mice against virulent S. aureus strains. The results obtained showed that this vaccine exhibits a broad spectrum of antigenic and immunogenic properties that protects mice from homologues and hetrologous S. aureus challenge.

Animals↗

Staphylococcus aureus strains isolated from bovine mastitis: virulence, antibody production and protection from challenge in a mouse model.

Septic arthritis in mice was used as a model to evaluate the virulence of Staphylococcus aureus and coagulase-negative staphylococci (CNS) isolated from cases of bovine mastitis. In addition, the model was used to evaluate the cross protection elicited by heterologous antibodies. Mice were intramuscularly inoculated with serial bacterial doses of different strains of S. aureus or CNS, for virulence determination; they were monitored for arthritis, gangrene or death up to 20 days. Antibody response, cross reactivity and resistance to challenge were tested by subcutaneous inoculation with a low dose of one of the S. aureus or CNS strains followed by challenge with two S. aureus strains. S. aureus alpha-hemolysin isolate was the most virulent, followed by alpha+beta-hemolysin and beta-hemolysin isolates. The least virulent isolates were the non-hemolytic S. aureus strains but even they were more virulent than the CNS strains tested. Antibodies against three different S. aureus antigens were detected by the ELISA in all mice that were inoculated with the S. aureus strains but not in any of those with the CNS strains. Immunoblot test against various S. aureus strains as antigens showed high cross-reactivity among the S. aureus strains but only a slight similarity, restricted to the bands above 36 kDa, with the CNS sera. Low-dose inoculation of alpha or alpha+beta strains before challenge with homologous and heterologous strains protected the mice, whereas the two beta strains provided only partial protection. The inoculations of non-hemolytic S. aureus or the CNS strains did not elicit any protection. Our findings demonstrate that pre-exposure of mice to a low dose of certain S. aureus strains could provide protection and that the antibodies produced could have an important protective role.

Animals↗

Immune cell differentiation in mammary gland tissues and milk of cows chronically infected with Staphylococcus aureus.

This study identifies and compares the distribution of mononuclear cells in the mammary gland tissues and milk of healthy and chronically infected with Staphylococcus aureus cows. Somatic cell counts (SCCs) during the 3 months before the study were > 1 x 10(6) cell/ml in the infected quarters and < 1 x 10(5) cell/ml in the infection-free quarters. Immediately after slaughter, samples from the tissues above the gland cistern and supra-mammary lymph node were collected. No histological differences were found between the supra-mammary lymph nodes of the healthy and infected udders, and both appeared normal. In the milk of the healthy infection-free mammary glands, SCC was < 50,000 cells/ml) while epithelial cells were the predominant type. The percentage of CD18+ was low than 45%, of which over three-quarters were polymorphonuclear (PMN), and less than one- quarter were mononuclear cells. The later comprised CD4+ or CD8+ T-lymphocytes, macrophages (Mo) but not B-cells. In the tissues, there were few CD18+ leukocytes, and most of the cells were T-lymphocytes. The number of B-lymphocytes bearing CD21+ was similar to that of CD8+ and were localized in the connective tissue as clusters of 2-5 cells, mainly in areas with no alveoli, or as single cell having a dendritic like form. The number of Mos was negligible. In the milk of the infected glands, SCC exceeded 700,000 cells/ml, of which > 95% were CD18+ positive. The distribution of the leukocytes had two patterns: one presented (> 80%) of PMN cells and a small number of mononuclear cells; the second had less than 50% PMN and many mononuclear cells. The CD8+ cells in these infected sections were observed throughout the mammary epithelial cells (MEc) around the alveoli and in the alveolar lumen (AL). The numbers and the location of CD21+ B-lymphocytes were similar to those in the infection-free mammary glands. The number of CD5+ positive cells was lower than T and B- cells combined and were located throughout the mammary epithelial cells, around the alveoli and within the connective tissue. Mo numbers were high in most of those infected quarters, and were localized around the connective tissue and within the AL.

Animals↗

Staphylococcus aureus exosecretions and bovine mastitis.

The role of Staphylococcus aureus and coagulase-negative staphylococcal exosecretions in bovine udder infection was tested by monitoring the cows' response to in vivo inoculation of bacterial exosecretions into udder quarters. Twenty Israeli-Holstein dairy cows were included in the study; two or three of the udder quarters of each cow were intracisternally inoculated with 0.04-0.05 mg/quarter (total proteins) of the various sterile bacterial exosecretions in a sterile pyrogen-free saline. Each udder was inoculated with two or three different bacterial exosecretions or placebo (Columbia Broth). Cows were monitored for 96 h post-inoculation for rectal temperature, heart and respiratory rates, alimentary tract activity (rumen contraction), udder temperature, pain, oedema and udder size. Milk samples were examined bacteriologically and for somatic cell count, N-acetyl-D-glucosaminidase (NAGase) activity and somatic cell differentiation. No enterotoxins (beta-G) or toxic shock syndrome toxin-1 were detected in response to any of the bacteria tested. Control quarters or those inoculated with Columbia Broth, showed similar NAGase and somatic cell count values throughout the experiment. Twelve of the 18 strains tested, induced inflammation in the inoculated quarters while six did not. Of the 12 strains causing local inflammation, only six were found significantly different from the control and were considered as high response (group 1). The other six that caused a local inflammation did not differ significantly from the control, and were considered to be moderate response (group 2). The six S. aureus isolates that did not cause an inflammatory response were considered to have low response (group 3). In all quarters inoculated with S. aureus bacterial exosecretions belonging to groups 1 and 2, the polymorphonuclear cells and macrophages were proportionally increased while CD4+ and CD8+ T-lymphocyte populations decreased. One-dimensional NuPAGE (7%) Tris-acetate gel electrophoresisof the bacterial exosecretions revealed four different bands appearing between 36 and 31 kDa, marked from top to bottom as A, B, C and D. An association was found between the combinations of expressed bands and the cow responses: the majority of the cases could be linked to the expression of bands B and C.

Acetylglucosaminidase↗

Infusions of casein hydrolyzates into the mammary gland disrupt tight junction integrity and induce involution in cows.

Milk stasis triggers local stimuli, which make the tight junctions leak and trigger involution. The aim of the study was to test the hypothesis that casein hydrolyzates compromise tight junction integrity and dry-off milk secretion in dairy cows. Six repeated doses of casein hydrolyzates after each milking during 3 d caused drastic changes in mammary secretion and composition, which were associated with irreversible cessation of milk secretion. No such changes were recorded in the control glands that had been treated with nonhydrolyzed casein. Treatment with casein hydrolyzates disturbed tight junction integrity within 8 h (as indicated by changes in Na+ and K+ concentrations), reduced the concentrations of lactose precipitously, activated the plasmin activator-plasminogen-plasmin system, and induced the secretion of immunoglobulin type G and lactoferrin. At the end of the 3-d treatments, we stopped milking the experimental and control glands. Milk composition 19 d later was similar in the experimental and control glands and was consistent with the composition expected in fully involuted glands. We conclude that casein hydrolyzates are among the milk-borne factors that cause the disruption of tight junction integrity and induce involution in cows. The process induced by casein hydrolyzate was more rapid and synchronized than the involution induced at drying-off.

Albumins↗

Autologous blood stem cell transplantation in refractory systemic lupus erythematosus with severe pulmonary impairment: a case report.

For patients with severe forms of autoimmunity, including systemic lupus erythematosus (SLE), purging autoreactive T cells from the immune repertoire by transplanting autologous hematopoietic stem cells (ASCT) is a therapeutic option. We describe an 18-year-old woman with SLE who had been treated with corticosteroids, azathioprine, cyclophosphamide (CYC), and immunopheresis for 4 years, during which time mechanical ventilation for lupus pneumonitis had been repeatedly required. After the patient was conditioned by administration of CYC and antithymocyte globulin, a total of 8.87 x 10(6) purified CD34+ cells per kg of body weight was infused. Hematopoietic regeneration was observed within 9 days. Twenty-one months after ASCT, the patient continues to be in complete clinical remission, with no signs of SLE-related disease activity and without any immunosuppressive medications. Her pulmonary function has returned to normal. Although a longer followup is required for assessment of the durability of response, the patient's course indicates that ASCT may be a way to reinduce tolerance in patients with SLE.

Adolescent↗

Influence of Staphylococcus aureus exosecretions isolated from bovine mastitis on leukocyte activity in vitro.

The role of Staphylococcus aureus and coagulase-negative staphylococcal exosecretions was tested for its ability to elicit in vitro proliferation of bovine blood lymphocytes, which we determined by means of the 3H-thymidine proliferation assay and by flow cytometry. Exosecretions of 32 field strains of S. aureus isolated from bovine udder infection and one of each of S. intermedius (M2), S. hyicus (M5), S. xylosus (M6) and S. chromogenes (M10) were used. Of the 32 S. aureus bacterial exosecretions, only 14 stimulated bovine mononuclear cells to proliferate. A high degree of association was found when the proliferation indexes were compared with the virulence as determined by intracisternal inoculation. All the six S. aureus strains that were categorized as highly virulent and that were tested in the proliferation assay exhibited a proliferation index > 20, whereas the five S. aureus strains that were categorized as low did not stimulate at all. Cells treated with media or Columbia broth supplemented with 0.1% D-glucose, yeast extract, and 0.5% NaCl (CBs) did not exceed 15% of the T-cells double positive with CD25+, whereas incubation with Con A activated the T-cells to display CD25+ up to 90%. Cells treated with one of the exosecretions that stimulated bovine mononuclear cells to proliferate, stimulated CD3+ and CD4+ T-cells to exhibit CD25+ receptor significantly higher (P < 0.05) than that found in media and CBs treatments, but lower than those found in Con A treatments. The exosecretions that did not stimulate mononuclear cells to proliferate also did not activate T-cells to exhibit CD25+ receptor. Con A activated 74% out of the total CD8+ to exhibit ACT2 receptor and 50% out of the total CD4+ to exhibit ACT3 receptor. A few but not all of the exosecretions that activated the CD25 receptor on T-cells also activated the ACT3 receptor on CD4+ cells.

Animals↗

A PCR-based method for the detection of Streptococcus agalactiae in milk.

Bovine mastitis caused by Streptococcus agalactiae is mainly subclinical and therefore can be diagnosed only in the laboratory. We developed a polymerase chain reaction (PCR)-based method for specific and sensitive detection of S. agalactiae in raw milk. The specificity of the PCR reaction is based on unique S. agalactiae DNA sequences within the 16S subunit of the rRNA genes. Two pairs of sequences were used as positive controls; general streptococci primers, which anneal to conserved areas within the 16S rRNA subunit gene, and primers, which anneal to sequences within bovine mitochondrial DNA. The method of detection includes selective enrichment of S. agalactiae in the milk sample, followed by DNA extraction using a rapid and simple procedure developed for this purpose, and specific PCR reaction with appropriate controls. The method enables the detection of one bacterium in 1 ml of raw milk. The method developed can be easily incorporated as part of routine screening of bulk milk collection tanks for early detection of infected cows in a herd.

Animals↗

Udder disease etiology, milk somatic cell counts and NAGase activity in Israeli Assaf sheep throughout lactation.

Bacterial pathogens causing udder infections in Israeli Assaf dairy sheep were identified and changes occurring throughout lactation were monitored to study the correlation between the contaminant and the severity of the infection, as measured by somatic cell count (SCC) and NAGase tests. A total of 159 Israeli Assaf dairy sheep on one farm, in their first (69), second (13) or third and more (77) lactations were included in this study. Udder halves were tested for bacterial condition, SCC and NAGase activity 2-3 weeks post lambing and every 4 weeks after until drying-off. At first sampling, in 60.7% (193/318 quarters) of the halves no bacterial growth (NBG) was detected. Different species of coagulase negative staphylococci (CNS) were the main pathogen group in infected udders. Streptococci were isolated from 14 halves, most of them in the two udder halves. The percent of udder infection in sheep in their third or further lactations was 2.8 greater (P<0.05) than in that of sheep in their first lactation. During the lactation, 90.6% of the halves did not change their classification status, suggesting that most infections occur before lambing and/or during the following first few days. The arithmetic mean of SCC and NAGase of total half udder milk and samplings (during the lactation) were 1144+/-48x10(3)cells/ml and 49.4+/-2.5, respectively. The average SCC in the milk of halves classified as NBG was 321+/-35x10(3)cells/ml and was not significantly changed during the lactation period. In halves infected with CNS, average SCC was 1371+/-150x10(3)cells/ml at the first testing and increased to 2129+/-347x10(3)cells/ml at drying-off. No significant differences were found in SCC and NAGase activity between the different species of the CNS. The mean SCC over the types of bacteria isolated, lactation number and days in lactation was significantly different (P<0.0001). In 4% of the halves, from all samples, SCC was above 5000x10(3)cells/ml although no bacteria were detected in their milk. The higher SCC in the CNS infected halves contrasted with the more moderate SCC found in dairy cows similarly infected, suggesting that the sheep udder has a lower resistance and an augmented immunological response against this group of bacteria. Thus, this should be considered accordingly in schemes for sheep's milk quality payment.

Journal Article↗

The effect of apramycin on colonization of pathogenic Escherichia coli in the intestinal tract of chicks.

The purpose of the present study was to examine the effect of apramycin sulphate on the colonization of pathogenic E. coli in the intestines of chicks. Apramycin treatment (0.5g/l in the drinking water) of 3-to 5-week-old Leghorn chicks for 24 or 48 hours resulted in a reduction, to an undetectable level, in the number of coliforms in the digestive tract for at least the first 24 h. Per os inoculation of E. coli (O2:K1) after 24 to 48 h of treatment resulted in a significant decrease in colony forming units (cfu) in the digestive tract of the treated chicks. Food deprivation from the time of inoculation did not significantly change the results. However, food and water deprivation caused bacteraemia in a number of the control chicks but not in the treated chicks. Comparison of the level of protection between Leghorn and broiler (Anak strain) chicks revealed that there was a significantly higher (P<0.05) level of bacteraemia in the broiler than in the Leghorn chicks. Chicks treated with 0.25 g/l or 0.125 g/l apramycin for 24 or 48 h before E. coli inoculation showed significantly lower cfu in the colon and caecum than untreated control chicks, but significantly higher cfu were found in the colon than in chicks treated with 0.5 g/l apramycin. Although in vitro preincubation of apramycin with ileum cells did not decrease the percentage of cells to which the bacteria adhered, the number of bacteria adhered per cell decreased significantly. Taken together, our in vitro and in vivo results show that apramycin is effective against E. coli by preventing colonization of the gut by the bacteria, which could lead to a reduction of colibacillosis in poultry.

Animals↗

Increased serum flt3-ligand in healthy donors undergoing granulocyte colony-stimulating factor-induced peripheral stem cell mobilization.

The effect of granulocyte colony-stimulating factor (G-CSF) induced mobilization of peripheral blood progenitor cells (PBPC) on the endogenous serum levels of cytokines stem cell factor (SCF) and flt3-ligand (flt3-L) was studied in 18 healthy subjects undergoing allogeneic PBPC donation. Donors received a standardized mobilization regime consisting of a 4-day course of G-CSF, with leukapheresis on day 5. Endogenous serum flt3-L and SCF were determined prior to G-CSF administration, on the day of leukapheresis, and followed up until day +100 after cessation of G-CSF administration. The administration of G-CSF resulted in a transient elevation of endogenous flt3-L serum levels. At the day of leukapheresis serum flt3-L showed a median increase of 75% compared to serum flt3-L levels obtained before G-CSF treatment. The increase in serum flt3-L levels showed no correlation with the total number of progenitor cells mobilized. Cessation of G-CSF treatment led to normalization of serum flt3-L within 7 days post G-CSF administration. In contrast, serum CSF levels remained unchanged in response to G-CSF administration. Our results demonstrate a transient surge in serum flt3-L in relation to G-CSF-induced PBPC mobilization, although the assessment of endogenous flt3-L give no information regarding the ability for G-CSF-induced PBPC recruitment in healthy individuals.

Adolescent↗

Immune response and resistance to infectious bursal disease virus of chicken lines selected for high or low antibody response to Escherichia coli.

Two experimental broiler lines were developed by divergent selection for high (HH) and low (LL) antibody response to Escherichia coli. Antibody response of these lines to immunization with a commercial vaccine (whole inactivated virus, WIV) against infectious bursal disease virus (IBDV) or with proteins VP2 and VP3 of that virus, and their resistance to challenge with a virulent IBDV, were tested. The study was performed with 213 male and female chicks from the tenth generation of the HH and LL lines. At 15 d of age, after disappearance of maternal antibodies, chicks from each line were randomly divided into four groups and injected with WIV, VP2, VP3, or adjuvant alone as a negative control. Chicks were bled 18 d postinjection, and antibody titers were determined by ELISA. Ten days later, the chicks were challenged with a virulent strain of the virus and killed after 10 d; the ratio of bursa of Fabricius to 100 g BW was determined for each bird. Significant differences in antibody titers were found among immunized and control chicks. Chicks from the HH line exhibited significantly higher antibody titers than LL chicks in response to WIV and VP2 vaccines but not to VP3 vaccine. Following challenge, bursa weight (relative to BW) of HH and LL chicks vaccinated with WIV and VP2 was significantly higher (P < 0.01) than that of chicks vaccinated with VP3 or the challenged unvaccinated control. No difference was found in this parameter between the latter two groups. Possible explanations for the differences in the line response to VP2 and VP3 are discussed.

Animals↗

Effect of gonadal steroids on proliferative responses and subset alterations in cultured chicken lymphocytes.

The effect of gonadal steroids (GS) on proliferation of lymphocytes and distribution of lymphocyte subpopulations in cell culture was examined. The involvement of protein kinase C (PKC) and calcium ionophore in the proliferative response was tested. Estradiol benzoate (EB) or testosterone propionate (TP) had no significant influence on proliferation of peripheral blood lymphocytes (PBL) when cells were not stimulated by mitogen. At high concentration (10-6 M), EB and dihydrotestosterone (DHT) decreased lymphocyte proliferative response to concanavalin A (ConA) and lipopolysaccharide (LPS) at 24 and 72 h of incubation. However, at physiological doses (10(-12) to 10(-16) M), EB significantly enhanced the proliferative response at 24 h of incubation, whereas DHT had no effect. The inhibitory effect of the high dose of EB or DHT on proliferation of T and B lymphocytes was independent of time of hormone presentation to the cells or age and gender of cell donor. In all cultures, pre-incubation of lymphocytes with 10(-6) M of EB or DHT significantly reduced their proliferative responses to ConA, phytohemagglutinin (PHA), and LPS. The percentage of CD3+ cells was significantly reduced by EB, whereas DHT had no such effect. In contrast to inhibition of proliferation in response to mitogens, 10(-6) M EB dramatically enhanced the proliferation of lymphocytes in response to the PKC activator, phorbol 12-myristate 13-acetate, and calcium ionophore, A23187. Results suggest that high doses of EB do not damage the viability or proliferation capability of lymphocytes and, therefore, suppress the proliferative response to mitogens in a different manner, perhaps by reducing gene transcription for receptors that recognize the mitogens, or suppressing some postreceptor events. The enhancement of proliferation in response to mitogens by low doses of EB may support this assumption, because the biphasic effects of steroids on gene transcription are well documented.

Animals↗