Pathophysiological effects of endotoxins in ruminants. 1. Changes in body temperature and reticulo-rumen motility, and the effect of repeated administration

Data from the literature on the clinical effects of bacterial endotoxins in ruminants are reviewed. Special attention is paid to the effects on body temperature and reticulo-rumen motility. Furthermore, the effects of repeated intravenous injection of endotoxin are summarised. Pathophysiological disturbances after intramammary infusion of endotoxins proved to be identical to those found after intravenous injection of non-lethal doses. Strikingly, however, no marked inhibitory effect on rumen motility nor abortion was observed after intramammary infusion of endotoxins. Moreover, in cows that were made tolerant to endotoxin by daily intravenous injections, intramammary infusion of one-fifth of this daily dose produced a maximum effect on body temperature and plasma Zn concentrations. This suggests that inflammatory endogenous mediators were released in the udder and then absorbed into the blood circulation, rather than the absorption of endotoxin.     

Systemic adverse reactions in young Simmental calves following administration of a combination vaccine.

Combination vaccines containing viral and bacterial antigens are commonly used in veterinary practice and have been associated with adverse reactions. A group of young Simmental calves developed fever and depression following administration of a mixed vaccine, and 1 died with pulmonary edema, suggesting that endotoxins or other bacterial components may interact synergistically with some adjuvants to cause an enhanced pathologic inflammatory response in some individuals.     

注射用盐酸土霉素的细菌内毒素检查

参照<中国兽药典>二○○○年版一部附录细菌内毒素检查法,研究了注射用盐酸土霉素对细菌内毒素检测的干扰情况,确定0.312 5 mg/mL浓度的供试品溶液对检测无干扰作用,建立了注射用盐酸土霉素的细菌内毒素限量检查方法.     

油剂普鲁卡因青霉素注射液细菌内毒素检查方法的研究

对油剂普鲁卡因青霉素注射液细菌内毒素检查方法进行了研究,在样品中加入适量的无菌、无内毒素的０．１ｍｏｌ／Ｌ氢氧化钠％吐温－８０溶液;并用０．１ｍｏｌ／Ｌ盐酸调节样品液的ＰＨ值,解决了样品的溶解方法,。用稀释法排除样品对细菌内毒素检查的干扰。实验结果表明,选用鲎试剂灵敏度为０．５ＥＵ／ｍｌ将样品稀释 １２５００ｕ／ｍｌ的溶液后可消除其干扰作用,因此,用细菌内毒素检查法作为该药品的热原物质检查是可行持     

The role of endotoxins in induced ruminal acidosis in calves.

Experimentally induced ruminal acidosis was carried out in 4 calves to investigate the possibility of resorption of endotoxins from Gram negative bacteria originating from the gastro-intestinal tract. The ruminal acidosis was induced by overfeeding of oats and the effect was evaluated by clinical and blood biochemical changes. Blood samples were collected every 2 h for 60 h before and after experimental feeding. The animals showed signs of ruminal acidosis and also the clinical and blood biochemical changes were similar to those seen after experimental endotoxaemia. However, although being less dramatic the changes in many relevant parameters such as an increase in prostaglandin F metabolite levels, body temperature, endotoxins and a decrease in iron indicate that an endotoxaemic state had occurred. The results of the present study show that ruminal acidosis/stasis is linked to resorption of endotoxins from the gastro-intestinal tract causing endotoxaemia/endotoxicosis.     

Bacterial translocation: clinical implications and prevention.

The occurrence of BT has been well documented in experimental animal models of hemorrhagic shock, trauma, severe burns, cirrhosis, pancreatitis, and bacterial overgrowth. Translocation of viable bacteria and endotoxins into mesenteric lymph nodes and other gut-associated lymphatic tissue is thought to activate a complex interplay of mediators that initiates the SIRS. Multiple humoral and cellular systems cause synthesis, expression, and release of inflammatory mediators, such as toxic oxygen radicals, proteolytic enzymes, adherence molecules, and various cytokines. A massive sustained proinflammatory response can ultimately result in irreversible multiple organ dysfunction. Because BT is associated with splanchnic hypoperfusion, the cornerstone of therapy involves rapid resuscitation and restoration of tissue perfusion. If a septic focus can be identified, it should be removed. Gut protectants, promotility agents, antioxidants, and immune-enhancing diets have shown promise in improving length of survival in these critically ill patients.     

Pathophysiological effects of endotoxins in ruminants. 2. Metabolic aspects

Metabolic disturbances following intravenous and intramammary administration of endotoxins in ruminants are described. In contrast to the similarity in response of blood biochemical parameters after intravenous and intramammary administrations of endotoxins, responses in plasma concentrations of enzyme activities, the thyroid hormones, cortisol, and somatotropin differ markedly. Biochemical changes in blood after endotoxin administration are predominantly dose-dependent; thus some of the biochemical parameters - especially plasma concentrations of Fe and Zn - serve also to evaluate the effects of certain drugs in endotoxin models. Changes in milk composition have been documented only after intramammary infusion of endotoxins and can partly be explained by the increased permeability of the blood/milk barrier. Appearance and production of milk returns to normal within a week after intramammary endotoxin treatment, indicating that the mammary gland is only temporarily damaged by endotoxin-induced mastitis.     

Concentration of airborne endotoxins and airborne bacteria in Chinese rabbit houses

Concentration of airborne endotoxins, airborne aerobic bacteria and airborne aerobic gram-negative bacteria were measured in 3 rabbit houses. Further, the species composition of the airborne gram-negative bacterial flora was investigated. The total amount of airborne endotoxin ranged from 22 to 774 EU/m3 (Endotoxin Units/m3). The number of total airborne aerobic bacteria varied between 780 and 20100 CFU/m3, the number of airborne aerobic gram-negative bacteria between 39 and 1030 CFU/m3. Most gram-negative bacterial isolates belonged to the family Enterobacteriaceae with E. coli as primary species. In two rabbit houses also airborne Pasteurella multocida spp. multocida, the most common respiratory pathogen of rabbits, was isolated.     

Up-regulation of NOD1 and NOD2 through TLR4 and TNF-alpha in LPS-treated murine macrophages

NOD1 (Card4) and NOD2 (Card15) are thought to be responsible for cytoplasmic defense against bacterial entry. To gain further knowledge about how their expressions are regulated in murine macrophages, we investigated the expression of NOD1 and NOD2 mRNAs after stimulation with various endotoxins, lipopolysaccharide, lipoteichoic acid and peptidoglycan. In macrophage RAW264.7 cells, the first and second rises in NOD1 and NOD2 mRNAs were observed at 2 hr and at 8-12 hr after endotoxin treatment. Increases in NOD1 and NOD2 mRNAs at 2 hr in lipopolysaccharide-treated RAW264.7 cells were reduced with the use of NF-kappaB inhibitor, caffeic acid phenethyl ester. In RAW264.7 cells, lipopolysaccharide-induced increases in NOD1 and NOD2 mRNAs were inhibited with anti-TLR4 antibody, and partially reduced in peritoneal macrophages obtained from TLR4-deficient mice. Furthermore, NOD1 and NOD2 mRNA expressions in RAW264.7 cells were increased by the treatment with proinflammatory cytokines, tumor necrosis factor-alpha (TNF-alpha), interleukin-1beta (IL-1beta), or IL-6. In TNF-alpha deficient macrophages, the expression of NOD molecules was minimal at 12 hr, and the second rise in NOD mRNA seen in lipopolysaccharide-treated RAW264.7 cells was inhibited with anti-TNF-alpha, but not with anti-IL-1beta or anti-IL-6 antibody. These observations suggest that immediate response of NODs to endotoxins could result from NF-kappaB activation via TLR signaling, whereas the second rise in NOD mRNAs might have resulted from TNF-alpha production possibly through NF-kappaB, TLR, and/or NOD signalings.     

注射用硫酸链霉素细菌内毒素检查法的探讨

按照《中国兽药典》二○○五年版一部[1]中注射用硫酸链霉素细菌内毒素检查法对样品复核检验,试验表明该方法是可行的;并与《美国药典》进行了比较,结果说明《中国兽药典》注射用硫酸链霉素细菌内毒素检查法对细菌内毒素的限值表达比《美国药典》29版[2]更合理、更科学。     

The effects of ascorbic acid on in vitro heterophil function

As a feed additive, ascorbic acid has been shown to have a protective effect against bacterial and viral diseases and to reduce the impact of detrimental stress in chickens. This study examined the effect of ascorbic acid treatment on in vitro heterophil function by examining random migration and phagocytosis and bacterial killing of Staphylococcus aureus. Heterophils were evaluated in broiler chickens ranging from 5 to 16 wk of age, and age differences were seen. Significant increases in bacterial killing were found in heterophils treated with ascorbic acid, and this difference tended to be greater in chickens from 5 to 10.5 wk of age. No significant differences were found in phagocytosis or random migration, but ascorbic acid tended to decrease random migration. The most significant effect on in vitro heterophil function was an increase in bacterial killing.     

Avian cathelicidins: paradigms for the development of anti-infectives

The broad-spectrum defense system based on host defense peptides (HDPs) is evolutionary very old and many invertebrates rely on this system for protection from bacterial infections. However, in vertebrates the system remained important in spite of the superposition of a very sophisticated adaptive immune system. The cathelicidins comprise a major group of HDPs in mammals. About six years ago it was first described that cathelicidins are also present in birds. Here we review the properties and biological activities of the recently discovered avian cathelicidins and their potential to be used as a paradigm for the development of anti-infectives. Like the mammalian cathelicidins, avian cathelicidins exert direct antimicrobial activities but can also selectively boost host immune responses by regulation of cytokine production and recruitment of immune cells. In addition, it was found that chicken cathelicidins bind endotoxins and dampen the endotoxin-mediated inflammatory response. Molecular dissection has allowed identification of different structural elements involved in bacterial killing and immunomodulation. These studies have enabled the design of small HDP-based antibiotics with specific functions, i.e. having primarily immunomodulatory or antimicrobial activities. Since the immunomodulatory effects may, to a certain degree, be species-specific, we hypothesize that poultry-specific antibiotics can be developed based on avian cathelicidins.     

The role of endotoxins in methscopolamine induced ruminal stasis in calves.

Methscopolamine was used to induce ruminal stasis in calves. Clinical and blood biochemical parameters were studied to judge the possible role of gastro-intestinal endotoxins from Gram-negative bacteria. Two trials were carried out where one injection of 100 mg and 3 consecutive injections of 70 mg of methscopolamine were administered. The animals showed signs of ruminal stasis. General clinical signs and changes in blood biochemical parameters were similar to what is found in endotoxaemia or in induced ruminal acidosis. Relevant parameters such as prostaglandin F2 alpha metabolite, endotoxin, iron, zinc, calcium and glutamate dehydrogenase changed significantly indicating exposure of endotoxins.     

Response of caeruloplasmin to Escherichia coli endotoxins and adrenal hormones in the domestic fowl

The injection of chickens with Escherichia coli endotoxins immediately produced a 50 per cent rise in plasma caeruloplasmin activity which was attributed to the release of the protein from liver cell. This was followed by a fall in activity, which was probably due to a fall in activity, which was probably due to a stabilising effect of adrenocortical hormones on the cell membranes, and then by a five-fold increase. The results of experiments with cycloheximide, adrenocorticotrophin, beta-methasone and reserpine indicated that the third phase of the response reflected increased synthesis in the liver which was partly induced by adrenal hormones. It increased with the dose and was not elicited by the particulate nature of the toxin preparation or by its lipid and polysaccharide components.     

Airborne gram-negative bacterial flora in animal houses

The concentration and the species composition of airborne gram-negative bacteria were studied in four cattle houses, one pig house and one poultry barn. On average only between 0.02 and 5.2% of the total number of culturable aerobic bacteria were identified as gram-negative bacteria. Obligate anaerobic gram-negative bacteria were not isolated at all. In the airborne gram-negative bacterial flora the following bacterial families dominated: the Enterobacteriaceae, the Pseudomonadaceae and the Neisseriaceae. Within the family of the Enterobacteriaceae the species Escherichia coli and Enterobacter agglomerans were predominant. In animal houses using straw as bedding material Ent. agglomerans was most frequent, whereas in animal houses without litter E. coli was mainly found. Airborne Neisseriaceae were isolated very frequently in cow barns with Acinetobacter lowffii as the primary species. Airborne Pseudomonadaceae were found in high concentrations during periods of high air humidity. The results presented may also give some indications on the origin and sources of airborne endotoxins in animal housing.     

Infection of gnotobiotic calves with Escherichia coli o157:h7 strain A84.

Six colostrum-deprived, hysterotomy-derived calves were maintained under sterile conditions and fed a milk replacer diet. At five days of age, five of the calves were dosed orally with 10(9) cfu of Escherichia coli O157:H7 strain A84. They were killed after, one, two, six, 12 and 24 days, and samples were taken for bacteriological and pathological examination. The sixth uninfected control calf was killed at seven days of age and matched samples were taken for pathological comparison. The animals remained normal throughout the observation period. Bacteriological data indicated a heavy bacterial load of strain A84 throughout the gastrointestinal tract but the bacterium was not found in liver, kidney or muscle. No evidence of 'attaching and effacing' lesions in the small or large intestine was found although there was a mild inflammatory response in the intestinal tract, consisting mainly of infiltrating eosinophils.     

Inverse association between endotoxin exposure and canine atopic dermatitis

The development of canine atopic dermatitis (CAD) may be related to exposure to mite allergens, bacterial endotoxin and/or fungal glucans. In this study, indoor exposure levels of house dust mite allergens, endotoxins and fungal glucans were measured to determine their possible association with CAD. A case-control study including adult Labrador retrievers with (n=28) and without (controls; n=65) CAD was conducted. Dust samples were collected from the living room floor and the bedding and coat of the dog and these were analyzed for house dust mite allergens Der p1 and Der f1, endotoxin and (1→3)-β-d-glucan levels. Dog owners were also required to return a questionnaire regarding their home characteristics. The endotoxin exposure level in the coats of dogs was significantly inversely associated with CAD (odds ratio 0.38; 95% confidence interval 0.15-0.97; P<0.05). No significant difference was found in exposure levels to house dust mite allergens and fungal glucans. The results indicated that endotoxin exposure is inversely associated with CAD, suggesting a protective effect of high indoor endotoxin exposure towards the development of the condition.     

生长肉兔细菌感染抗病力的遗传变异

细菌感染是造成断奶后肉兔重大经济损失的疾病之一。本研究通过对4个品种787只5~10周龄断奶肉兔健康记录的分析,确定细菌感染疾病抗病力的遗传方差。利用父性阈值模型分析断奶后肉兔细菌感染疾病的发病情况。结果表明:细菌感染的发病率从第5周的最小值逐步增加到第9周的最大值。品种和父性效应对生长肉兔细菌感染的发病率影响显著(P<0.05)。比利时兔的发病率最高(36.87%),父性方差占到了总表型方差的3.42%~8.02%。因此,肉兔细菌感染疾病的抗病力存在遗传变异,通过遗传选择可以降低细菌感染的发病率。     

Age-stratified validation of an indirect Salmonella Dublin serum enzyme-linked immunosorbent assay for individual diagnosis in cattle.

Enzyme-linked immunosorbent assays (ELISAs) are routinely used for cattle herd diagnosis of Salmonella Dublin infection in many countries. It is also possible to use such tests for individual diagnosis. Passively transferred immunoglobulins may cause false-positive test results in young calves. Also, false-positive test results may be seen in recovered animals several months after infection. False-negative results are seen in acutely infected animals, especially immature animals that are unable to produce a humoral antibody response to infection. To be able to interpret the individual animal test results, it is necessary to take age into account when validating the ELISA. In the present study an age-stratified validation of an indirect Salmonella Dublin serum ELISA as predictor of bacterial excretion was performed using receiver-operating characteristic (ROC) analysis. Three age groups were formed according to the results of an exploratory analysis of the age effect on area under curve (AUC) of ROC curves. The AUC for the youngest age group (0-99 days) was 0.816 (SE = 0.033), which was significantly (z = 4.23, P < 0.0001) smaller than the 0.977 (SE = 0.019) estimated for the next age group (100-300 days). The oldest age group (> 300 days) had an AUC of 0.905 (SE = 0.023), which was significantly different from the AUC of both the other age groups (z = 2.21, P = 0.027 when compared with the youngest age groups and z = 2.41, P = 0.016 when compared with the age group of 100-300 days). The results showed that the indirect Salmonella Dublin serum ELISA is most valid for detection of infection in individual cattle from the age of 100 days. Purpose-related test sensitivities and specificities were evaluated at different cutoff values.     

Treatment of endotoxaemia and septicaemia in the equine patient

Endotoxins, constituents of the cell wall of gram‐positive and gram‐negative bacteria, regularly result in severe illness and death in horses. In endotoxaemia, these constituents are present in the systemic circulation; in septicaemia, whole microbes invade normally sterile parts of the body. Interaction of these endotoxins with pathogen recognition receptors leads to an inflammatory response that cannot always be sufficiently contained and hence needs direct treatment. Over the last decennia, our understanding of the pathophysiology of endotoxaemia and septicaemia has significantly increased. Based on improved understanding of the interaction between receptors and endotoxins as well as the subsequent downstream signalling pathways, new therapeutic targets have been identified in laboratory animal species and humans. Important species differences in the recognition of endotoxins and pathogens by their receptors as well as the inflammatory response to receptor activation hamper extrapolation of this information to the horse (and other species). Historically, horses with endotoxaemia and septicaemia have been treated mainly symptomatically and supportively. Based on the identified therapeutic targets, this review describes the current knowledge of the treatment for endotoxaemia and septicaemia in the horse with reference to the findings in other animal species and humans.