Vitamin and trace mineral content in feed of breeders and their progeny: effects of growth, feed conversion and severity of malabsorption syndrome of broilers.

1. A study was conducted to investigate the effects of several vitamins and trace elements chickens and in chickens experimentally infected with malabsorption syndrome (MAS). 2. Vitamins and trace minerals in feed were varied. Breeders received either a basal amount of vitamins and trace minerals (low mix) or an increased amount (high mix). Their progeny also received either a low mix or a high mix. Effects of different breeder and broiler mix combinations on broiler performance, heamatology, spleen weight and humoral response were examined in control chickens. The effects of the different feeds and breeder, broiler combinations at the severity and recovery of MAS infection were also studied. 3. In general, the immune system can be stimulated by addition of vitamins and trace minerals, without affecting the growth potential of the controls. The number of leukocytes increased on d 1 in the broilers descended from breeders receiving high mix. The response to Newcastle disease virus boost was affected by the different amount of vitamins. 4. When breeders received a high mix the number of infiltrating polymorphonuclear leukocytes in the intestine was higher compared with breeders receiving basal amounts of minerals and vitamins. Also the recovery rate of intestinal lesions, cystic crypts of Lieberkühn and villus atrophy, as observed by histopathology, was faster in the groups where the breeders received high mix.

Physiological effects of elevated plasma corticosterone concentrations in broiler chickens. An alternative means by which to assess the physiological effects of stress.

Research on physical or psychological stress, in order to monitor objective parameters for animal welfare, is usually performed during experimental stress induction. To avoid treatment of animals with physical or physiological stress, addition of the stress-related hormone corticosterone to the drinking water, may serve as a practical alternative to reproducibly investigate hormone-related stress in broiler chickens. Rapid uptake of the hormone and distribution in the bloodstream were affirmed by elevated plasma corticosterone concentrations immediately after start of the treatment. The effect of hormone administration was evaluated by examination of corticosterone-sensitive organs. Comparable to the observations during physiological stress, we found in our model that uptake of endogenous corticosterone reduced body and spleen growth, increased heterophil counts, and decreased formation of antibodies against sheep red blood cells. Furthermore, corticosterone decreased adrenal gland responsiveness, measured by corticosterone production, after a challenge with adrenocorticotropic hormone. The simple performance, and the close relation between circulating corticosterone levels and heterophil counts, makes this an easy and quick method that is sensitive to increased levels of circulating corticosterone from base levels. The changed responsiveness of the adrenal glands to adrenocorticotropic hormone after increased circulating corticosterone levels may be an indication of the coping strategies during stress. Therefore, this test may be a promising tool in the research of adaptation to stress by broiler chickens.

Automated blood cell count: a sensitive and reliable method to study corticosterone-related stress in broilers.

In chickens the heterophil/lymphocyte ratio (H/L) has proved to be a valuable tool in stress related research. In general, H/L is determined with the microscopic differential count on a blood film. We evaluated automated analysis for measuring blood cell parameters in relation to corticosterone in a recently introduced corticosterone model. Discrepancies between microscopic and automated counts were found for the percentage of monocytes and basophils. The relative H/L ratio appeared to be sensitive for increased plasma corticosterone levels. However, the increase in heterophil frequencies measured with the hematology analyzer proved to be the most sensitive method for the measurement of changes in plasma corticosterone concentrations. We therefore propose automated hematological analysis as a simple and sensitive tool to study the effects of physiological corticosterone concentrations on blood cell parameters in relation to stress in broiler chickens.

Survival and resuscitation of ten strains of Campylobacter jejuni and Campylobacter coli under acid conditions.

The culturability of 10 strains of Campylobacter jejuni and Campylobacter coli was studied after the bacteria were exposed to acid conditions for various periods of time. Campylobacter cells could not survive 2 h under acid conditions (formic acid at pH 4). The 10 Campylobacter strains could not be recovered, even when enrichment media were used. Viable cells, however, could be detected by a double-staining (5-cyano-2,3-ditolyl tetrazolium chloride [CTC]-4',6'-diamidino-2-phenylindole [DAPI]) technique, demonstrating that the treated bacteria changed into a viable but nonculturable (VBNC) form; the number of VBNC forms decreased over time. Moreover, some VBNC forms of Campylobacter could be successfully resuscitated in specific-free-pathogen fertilized eggs via two routes, amniotic and yolk sac injecting.

Development of organs and intestinal mucosa leukocytes in four broiler lines that differ in susceptibility to malabsorption syndrome.

Growth retardation in young broiler chicks due to poor nutrient metabolism, commonly known as malabsorption syndrome (MAS), is a widespread problem caused by enteric infections with a combination of pathogens mainly viruses. Genetic lines of broiler chickens differ in susceptibility to the syndrome. A difference in growth retardation was observed among four broiler lines (BL) after oral inoculation at 1 d of age with intestinal homogenates obtained from MAS-affected birds. Two of the lines that are more susceptible to MAS had severe weight gain depression. To uncover the factors that play a role in the susceptibility to MAS, we analyzed the growth rate of the body and vital organs and the quantity of leukocytes in the peripheral blood and intestinal mucosa. The development of the intestine, liver, bursa of Fabricius, and spleen was similar among the BL. The resistant BL had higher numbers of peripheral blood leukocytes, especially lymphocytes, at 1 d of age. A significant difference was noted in the numbers of CD4+ T cells and CD8+ T cells in the intestinal villi. At the ages of 3 and 8 d, the susceptible BL had more CD8+ T cells in the villi, whereas the ratios of CD4+:CD8+ T cells were higher in the resistant BL. This difference in the number of T-cell subpopulations in the intestinal mucosa might be an important factor in the difference in susceptibility to the enteric infections associated with MAS.

Experimental reproduction of malabsorption syndrome with different combinations of reovirus, Escherichia coli, and treated homogenates obtained from broilers.

Attempts to reproduce malabsorption syndrome (MAS) by oral inoculation with several different combinations including intestinal homogenate, reovirus, and hemolytic Escherichia coli obtained from MAS-affected chickens and intestinal homogenate from healthy chickens (healthy homogenate) were performed in 1-day-old specific-pathogen-free (SPF) broilers. The MAS homogenate, serving as a positive control, induced weight gain depression and intestinal lesions such as cystic crypts of Lieberkuhn, villus atrophy, and lymphoid and/or granulocytic infiltration. The healthy homogenate, the formalin-treated MAS homogenate, the formalin-treated healthy homogenate, and phosphate-buffered saline caused neither weight gain depression nor intestinal lesions. We were able to reproduce both weight gain depression and intestinal lesions by inoculation of reovirus either combined with the formalin-treated MAS homogenate or combined with healthy homogenate. Surprisingly, when hemolytic E. coli was added to the combination of reovirus with formalin-treated MAS homogenate, this did not cause weight gain depression although this combination caused the described intestinal lesions. Identical results were obtained with the combination of formalin-treated MAS homogenate with hemolytic E coli or the combination of reovirus with hemolytic E. coli. The intestinal lesions were more severe and developed faster by combinations including reovirus and formalin-treated MAS homogenate. This study indicates that a combination of enteropathogenic reovirus with other agents or substances that are present in an intestinal homogenate from MAS-affected and healthy chickens can induce MAS in SPF broilers. Escherichia coli is not essential for induction of weight gain depression but can play a role in development of intestinal lesions. Furthermore, intestinal lesions alone will not always result in weight gain depression.

Cellular immune response in the small intestine of two broiler chicken lines orally inoculated with malabsorption syndrome homogenates.

We studied the cellular immune response against malabsorption syndrome (MAS) in two broiler chicken lines, A and B. We determined the number of pan T-lymphocytes (CD3), helper T-lymphocytes (CD4), cytotoxic T-lymphocytes (CD8) and macrophages/monocytes in the small intestine in the first 2 weeks after oral inoculation of two MAS homogenates, MAS80 and MAS97-1. The immune cells were detected on cryostat tissue by immunohistochemistry and counted by villus area. In trial 1, we compared the two broiler lines for weight gain depression, intestinal lesion and number of CD3, CD4, CD8 cells and macrophages/monocytes after MAS80 inoculation. Although there was no significant difference in weight gain depression between the two broiler lines, line B had significantly higher numbers of CD8+ T-cells per villus area than had line A. To confirm part of the results of trial 1, trial 2 was done in which we compared different homogenates in broiler line B. Broiler line B was orally inoculated with either MAS97-1, intestinal homogenate obtained from healthy chickens (healthy homogenate), or phosphate buffered saline (PBS). In this trial, the MAS97-1 homogenate also induced weight gain depression and intestinal lesions, whereas the "healthy homogenate" and PBS did not induce weight gain depression or intestinal lesions. The broilers inoculated with MAS97-1 homogenate had significantly more CD8+ T-cells per villus area than had broilers inoculated with "healthy homogenate" or PBS. Increased CD8+ T-cells per villus area in the affected small intestines of broilers suggests an increase of cytotoxic T-cell activity.