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Biomedical subjects

S Mallet

Publications and source records attributed to S Mallet.

At least 19 recordsLinked to original sources

Effects of beak trimming and cage design on laying hen performance and egg quality.

1. Conventional cages are to be replaced by furnished cages or aviary systems to improve the welfare of hens. We compared the performance and egg quality of hens reared in two designs of furnished cages and of two standard cages. We also explored the consequences of the absence of beak trimming when using these designs. 2. Hens (2028) were housed from 18 to 70 weeks of age in 108 standard cages at 6 per cage (60 cmx63.5 cm), in 96 cages at 5 per cage (59.5 cmx55.5 cm) or in two designs of furnished cage at 15 per cage (24 F15M cages and 36 F15P cages made by two manufacturers) which contained equipment varying in size and location (nests, dust baths and perches). Half of the hens were beak-trimmed in each design. 3. Mortality was low in beak-trimmed hens (<5%) but was unacceptably high in non-beak-trimmed hens due to cannibalism (>40%, 516 hens). Mortality was worse in standard cages than in furnished cages. Consequently, hen-housed egg production was significantly lowered in non-beak-trimmed hens. 4. Egg laying in beak-trimmed hens reared in furnished cages occurred mainly in the nest (80 and 84.8% in F15M and F15P) but also in the dust bath (13.3 and 9.4% in F15M and F15P) and in other parts of the cage (6.7 and 5.8% in F15M and F15P). 5. The total percentage of broken (visual observation) and hair-cracked eggs (candling) was high in the furnished cage designs (15.4 and 19.6% in F15M and F15P, respectively) compared with standard cage designs (8.1 and 12.2% in S6 and S5). This was mainly due to hair-cracked eggs, the highest percentages occurring in the nests, especially in the design with a narrow nest and no egg saver (11.1% in F15M compared to 17.6% in F15P) as a consequence of egg accumulation in the cradle and relatively low frequency of manual egg collection. 6. Eggshell quality (index and breaking strength) was only slightly influenced by cage effects so differences in egg breakage were attributable to impacts related to cage design. 7. It is concluded that beak trimming remains the most effective way to prevent cannibalism, although furnished cages with a large group of hens slightly reduced the incidence, and that further development and optimisation of furnished cages is needed to reach egg quality similar to standard cages.

Animal Husbandry↗

Comparison of eggshell hygiene in two housing systems: standard and furnished cages.

1. The hygienic properties of eggs produced in two designs of furnished cage were investigated and compared to two standard cage designs. 2. At 28, 37, 47 and 58 weeks of age, the proportion of dirty eggs was higher in one of the furnished cage designs while, in the other, it was similar to standard cages. 3. At 27, 33 and 60 weeks, the bacterial load on the eggshell (total aerobic bacteria and enterococci) was higher in furnished cage designs. A seasonal effect was observed with lower counts at 60 weeks (winter) than at 27 weeks (summer). 4. More dirty eggs and a higher bacterial load were observed in eggs laid outside the nests, which suggests egg hygiene in furnished cages could be similar to standard cages if the equipment in furnished cages was improved to enhance nest laying.

Aging↗

Differences in the digestive tract characteristics of broiler chickens fed on complete pelleted diet or on whole wheat added to pelleted protein concentrate.

1. The effects of whole grains of wheat on the digestive tract of broiler chickens was studied. A complete pelleted feed was compared with free choice feeding of whole wheat and a pelleted protein concentrate, given from 7 to 29 d of age. 2. Pepsin activity in proventriculus tissue was lower in whole wheat-fed birds than in complete diet-fed birds. The weight (g/kg body weight) of the gizzard was higher in whole wheat-fed birds and its contents had a lower pH. 3. In the intestine, there were no differences in deoxyribonucleic acid (DNA) concentration, protein/DNA, ribonucleic acid (RNA)/DNA, RNA/protein ratios or alkaline phosphatase activity expressed per tissue weight. The weight (g/kg body weight) of the duodenum was lower in whole wheat-fed birds and its contents had a higher pH. Also the activities of alkaline phosphatase and leucine aminopeptidase in the duodenum, and maltase in the ileum, expressed per unit of bird weight, were lower in whole wheat-fed birds. 4. These results suggest that whole grain feeding increases the chemical (pepsin in proventriculus) and physical (gizzard muscle) functionality of the upper part of the digestive tract but decreases the digestive capacity of the intestine. Higher gizzard functionality may play a positive role in the control of bacterial populations. The lower digestive enzyme activities in the intestine may be detrimental in situations of mucosal deterioration caused by intestinal disease.

Animal Feed↗

Effects of whole wheat feeding on the development of coccidial infection in broiler chickens.

A complete ground and pelleted feed was compared to free choice feeding of whole wheat and a pelleted protein concentrate during three experimental infections with coccidia in broiler chickens. At 22 d of age birds were inoculated with different doses of sporulated oocysts of a cecal species (Eimeria tenella) in experiment 1 or intestinal species E. maxima or E. acervulina in experiments 2 and 3, respectively. The effects of diets were assessed on weight gain, hematocrit (during cecal coccidiosis), serum coloration (during intestinal coccidiosis), oocyst excretion, and lesion score until 7 d post-inoculation. In experiment 1 before inoculation, the birds fed whole grain had more beneficial microflora with lower counts of coliform bacteria. As shown by oocyst output and lesion score, whole wheat feeding increased parasite development during infection with the cecal parasite E. tenella. This led to significantly lower weight gain with whole wheat than with ground wheat from 5 to 6 d post-inoculation and to lower hematocrit at the highest infective dose. Parasite development in experiments 2 and 3 was similar among diets, during intestinal infection with E. maxima and E. acervulina, respectively, with no significant differences in lesion score. During the acute phase (4 to 7 and 3 to 5 d post-inoculation), when a difference appeared between diets, whole wheat fed-birds were always more affected than ground diet-fed birds in terms of serum coloration and weight gain. These results might be explained by modifications of digestive physiology and intestinal microflora by whole wheat.

Animal Feed↗

Effects of the initial level of milk production and of the dietary protein intake on the course of natural nematode infection in dairy goats.

The level of dietary protein is known to alter the establishment and the pathophysiology of gastrointestinal nematodes in sheep. On the other hand, high-producing dairy goats are less resistant and/or resilient than low-producing ones to experimental nematode infection. During a 2-year study, we have investigated the course of a natural nematode infection (mainly T. colubriformis) in a high-producing dairy goat flock. In year 1, 50 grazing goats divided in high (HP) or low producer (LP) were compared from April to October for parasitological and milk parameters. In year 2, the 25 HP goats were only considered and were allocated to two levels of dietary protein, high level (HPr) with a protein coverage of 125% or normal level (NPr) with a protein coverage of 106%. They were monitored as above. In year 1, HP goats showed a greater nematode egg output (1856 vs. 1000epg) associated with higher values in T. colubriformis IgG in autumn than LP ones whereas the decrease in serum phosphate concentration was similar in both groups. In year 2, HPr goats exhibited lower egg output in autumn (2219 vs. 2817) vs. NPr ones. Moreover, milk yield and fat content were significantly higher in HPr vs. NPr in the 2nd part of the study. HP goats are less resistant to nematode infection in natural conditions. Resistance and resilience of HP goats may partially be improved by a protein supplementation in the diet.

Animal Feed↗

Determination of tartaric acid in solid wine residues by capillary electrophoresis and indirect UV detection.

Tartaric acid, used in pharmaceuticals and industrial food preparation, is an important by-product of wine preparation. It is produced by wine factories in large quantities and cannot be rejected into the environment. Wineries precipitate tartaric acid using calcium hydroxide and then evaporate the mixture. The raw compact powder obtained, which contains calcium tartarate and a lot of other constituents (sugars, tannins, etc.) is sold to factories which purify tartaric acid. The different analytical methodologies which are used to determine the tartaric acid concentration in the solid wine residues are long and tedious (Goldenberg method, 14 samples per day). They also suffer from poor reproducibility. Some other methods use ion chromatography or solid Fourier transform IR, which are not currently used for such topics. We propose a capillary electrophoresis method, which not only is very quick (2 min of analysis) but also highly reproducible. Finally it provides very simple electropherograms. The intra-day and inter-day repeatability, the inter-person reproducibility and recovery were estimated. They demonstrate the ruggedness of this new method.

Benzoates↗

Effects of the excretory/secretory products of Trichostrongylus colubriformis on the growth of different cell lines.

The effects of the excretory/secretory (ES) products of the parasitic nematode Trichostrongylus colubriformis were examined on the proliferation of seven cell lines derived from a digestive or non-digestive origin. The excretory/secretory products of T. colubriformis were incorporated in the culture medium of the different cell lines and cell proliferation was measured by means of the 5-bromo-2'-deoxy-uridine (Brdu) assay. An increase in cell numbers was found with the three epithelial intestinal cells (RIC, IEC-6, IRD-98) and with epithelial kidney cells (MDCK). In contrast, an inhibition in the proliferation of epithelial ovarian cells (CHO) and fibroblasts (3T3) was observed with the addition of the excretory/secretory products and no effect was detected on the cell growth of hepatocytes (HepG2). These data are discussed with respect to the tissue specificity of the existing mitogenic effect of the worms on the intestinal crypt cells during parasitism.

3T3 Cells↗

Role of acetylcholinesterase (AChE) secreted by parasitic nematodes on the growth of the cell line from epithelial origin HT29-D4.

The excretory-secretory (E-S) products of the parasitic nematodes Trichostrongylus colubriformis and Nematodirus battus were found to modify the in vitro proliferation of the tumorous colic HT29-D4 cell line of epithelial origin. A characteristic feature of these E-S products is the presence of a high level of acetylcholinesterase (AChE) activity, the biological significance of which remains unclear. To determine a possible role of AChE on cell growth, the enzyme was purified from E-S products using edrophonium chloride. Purity was confirmed by polyacrylamide gel electrophoresis, using silver and Karnovsky stains, before assessing its effects on cell proliferation. The purified AChE was incorporated at different concentrations in a culture medium of HT29-D4 cells. A mitogenic effect was shown for low concentrations (0.1-14 units). By contrast, an inhibitory effect was noted at high concentrations (35-1400 units). Furthermore, polyclonal antibodies were prepared and depletion of AChE in E-S products by immunoprecipitation or affinity chromatography resulted in a partial or total disappearance of the stimulatory effect of cell growth. Thus, the results form this in vitro study suggest a modulatory role for AChE secreted by nematode parasites on the proliferation of epithelial cells of the host.

Acetylcholinesterase↗

Typing of Staphylococcus epidermidis strains by random amplification of polymorphic DNA.

The polymerase chain reaction was used to obtain randomly amplified polymorphic DNA profiles for typing of Staphylococcus epidermidis strains. Epidemiologically unrelated S. epidermidis isolates were screened with randomly amplified polymorphic DNA analysis. The discriminating ability of 45 randomly designed 10-mer primers was assessed. The highest discriminatory power was obtained with the 10-mer oligonucleotide OPAM-12. In typing a total of 13 unrelated S. epidermidis strains with OPAM-12, 11 different banding profiles were obtained reproducibly by agarose gel electrophoresis. The discriminatory power of the method with OPAM-12 was estimated using the D value of Hunter and Gaston (1988) to be 0.961. A reproducibility index of 1 was obtained after typing a total of 40 cultures including 12 triplicates and one quadruplicate of the 13 unrelated strains. Following the described procedure, the randomly amplified polymorphic DNA method provided a rapid, simple and reproducible alternative to other S. epidermidis typing systems.

Base Sequence↗

Physiology of two strains of Trichostrongylus colubriformis resistant and susceptible to thiabendazole and mucosal response of experimentally infected rabbits.

Two strains of Trichostrongylus colubriformis of ovine origin, one resistant and one susceptible to thiabendazole, were compared during experimental infection in rabbits. Groups of rabbits were infected with 5000 infective larvae (L3) of either the resistant or the susceptible strain. On days 21 and 42 post-infection (p.i.), 5 rabbits of each group were killed and the small intestine divided into three sections for worm counts and histological or biochemical analysis. Faecal egg counts were performed twice a week from day 15 to day 42 p.i. The physiology of the worms was characterised by in vitro acetylcholinesterase secretion of adult worms. The host inflammatory response was determined by peroxidase activity in mucosal homogenates and by histological counts of mast cells and eosinophils. Infectivity was not significantly different between the two strains. However, egg production was delayed and was significantly lower for the resistant strain. Conversely, in vitro secretion of resistant worms was significantly higher than the susceptible ones. The mucosal homogenetates of rabbits infected with the resistant strain had significantly higher levels of peroxidase activity in the duodenum on days 21 and 42 p.i. Mast cell counts were also significantly higher in the duodenum on day 42 p.i., in rabbits infected with the resistant strain. No difference was observed in the eosinophil counts. These results suggest that genetic variation in the nematode, such as anthelminthic resistance, is associated with variations in worm biology and physiology as well as differences in the inflammatory response of the host.

Acetylcholinesterase↗

Trichostrongylus colubriformis infection in rabbits: persistence of the distal adaptive response to parasitism after anthelmintic treatment.

Mucosal changes associated with anthelmintic treatment in Trichostrongylus colubriformis-infected rabbits were assessed along the entire length of the small intestine. The following groups, each of five rabbits, were compared: infected (group I); infected and treated on day 21 with fenbendazole (group IT); uninfected but given fenbendazole on day 21 (group C, controls). All animals were killed on day 28. In the proximal part of the small intestine of group I rabbits, the worms were associated with shortening of the villi and a significant depletion in alkaline phosphatase activity, which differed significantly from the findings in groups C and IT. In the same region, no difference was found between groups C and IT. In the distal small intestine, hypertrophy of villi and crypts (an adaptive response to the infection), coupled with an increase in enzymic activity, were present in both groups I and IT, in contrast to group C. These results suggest that a complete mucosal restoration occurred within 7 days of anthelmintic treatment in the parasitized part of the intestine. In contrast, the adaptive response observed beyond the main site of parasitism was not abolished by treatment. The functional significance of these findings is discussed in relation to the compensatory growth commonly observed after anthelmintic treatment in ruminants.

Adaptation, Physiological↗

Stimulation of HT29-D4 cell growth by excretory/secretory products of the parasite nematode Trichostrongylus colubriformis.

The presence of the nematode parasite Trichostrongylus colubriformis in the small intestine is associated with an increase in epithelial renewal. To assess the possible role of excretory/secretory products from the worm on cell proliferation, adult Trichostrongylus colubriformis were incubated in vitro in Dulbecco's Modified Eagle's Medium for 24 h and the conditioned medium was added to the culture medium of the transformed epithelial cell line HT29-D4. A stimulation of the HT29-D4 cell growth was ascertained at concentrations of 0.25-1.0 micrograms protein/ml using counts of cell numbers, the MTT method and incorporation of tritiated thymidine. An increased incorporation of tritiated thymidine was also observed with the excretory/secretory products from T. colubriformis fourth stage larvae at 1.0 microgram/ml. Dialysis of the medium conditioned by the worms indicated that the molecular weight of the factor is greater than 8000 Daltons in size. Heat treatment, acid hydrolysis and precipitation by trichloracetic acid of the conditioned medium resulted in the disappearance of the proliferative effect while treatment with trypsin partially depleted the stimulative activity. These results suggest that T. colubriformis produce some protein factor which could increase the epithelial regeneration in the host small intestine.

Animals↗

Effects of the excretory/secretory products of six nematode species, parasites of the digestive tract, on the proliferation of HT29-D4 and HGT-1 cell lines.

The excretory/secretory (ES) products of the nematode parasite Trichostrongylus colubriformis have been found to increase the in vitro proliferation of the epithelial cell line HT29-D4. To assess the specificity of this effect, ES products from other trichostrongyle species were tested on colonic (HT29-D4) and gastric (HGT-1) tumour cell lines. Adult worms of six different nematode species, parasites of the stomach or the small intestine of ruminants, were incubated in vitro in Dulbecco's Modified Eagle's Medium for 24 h. The conditioned media were then added at different concentrations to the culture medium of the two cell lines. A stimulation of the HT29-D4 cell growth occurred with the ES products of two parasite species of the small intestine, at the concentrations of 0.1 microgram protein/ml (Trichostrongylus vitrinus) and 1.0-5.0 micrograms/ml (Cooperia curticei). Inversely, a decrease in cell number was observed with the ES products of another intestinal species, Nematodirus battus at concentrations of 1.0-5.0 micrograms/ml. With the ES products of the abomasal nematodes, a proliferation of HT29-D4 cells was obtained at 0.25-5.0 micrograms/ml with ES products of Teladorsagia circumcincta but no significant effect was observed for Haemonchus contortus. On the tumoral gastric cell line HGT-1, the ES products from the 6 nematode species gave a similar stimulative effect. These in vitro results suggest that nematode parasite species secrete or excrete component(s) which could affect the epithelial regeneration of the host digestive tract.

Animals↗

Lack of intraspecific variation in the second internal transcribed spacer (ITS-2) of Trichostrongylus colubriformis ribosomal DNA.

The nucleotide sequence of the second internal transcribed spacer (ITS-2) was determined for three populations of the parasitic nematode Trichostrongylus colubriformis which differed in their susceptibility to benzimidazole anthelmintics and/or in their geographical origin. No intraspecific variation was found in the ITS-2 sequence, indicating that this region of rDNA is inadequate to discriminate between resistant and susceptible populations of T. colubriformis, but it may prove useful for distinguishing between species of Trichostrongylus.

Animals↗

Histopathology of the small intestinal mucosa in Nematodirus spathiger infection in rabbits.

Rabbits were experimentally infected with two levels (5000 and 17,000) infective larvae of Nematodirus spathiger. Histological (villus length, mucosa to serosa ratio, crypt surface) and biochemical (protein content, alkaline phosphatase and leucine aminopeptidase activities) measurements relating to the small intestinal mucosa were examined along the entire length of the organ. In the proximal intestine, the presence of worms was associated with villus abrasion, increased crypt surface and decreased alkaline phosphatase and leucine aminopeptidase activities. Conversely, beyond the main site of infection in the distal small intestine, some signs of hypertrophied villi and crypts were noted without any changes in enzyme activities. These distal variations were similar to those previously described in experimental Trichostrongylus colubriformis infections of rabbits. These results tend to confirm the use of the rabbit as an experimental model to study Nematodirus infection. They also suggest that the distal adaptive process in the nematode-parasitized small intestine could occur independently of the worm species.

Alkaline Phosphatase↗

The effect of levamisole on acetylcholinesterase secretion by male and female Heligmosomoides polygyrus during ageing.

The inhibition of acetylcholinesterase secretion by male and female Heligmosomoides polygyrus was tested on worms taken from experimentally infected mice and maintained for 3 days in vitro in levamisole. The dose inhibiting 50% of enzyme secretion (ID 50) of male worms was twice the ID 50 for female worms. A similar difference was observed in vivo between the dose of levamisole removing 50% (LD50) of male and female worms from the mouse. Acetylcholinesterase secretion by worms and ID 50 were tested in vitro at 3-weekly intervals from 3 to 21 weeks post infection (WPI). Acetylcholinesterase secretion was always significantly higher for male than for female worms. A decrease of ID 50, correlated with the age of the worms was observed: from 1.5 to 0.5 micrograms/ml for males and from 0.7 to 0.1 micrograms/ml for females. These results were confirmed in vivo by a higher efficacy of the anthelminthic at 21 than at 4 WPI.

Acetylcholinesterase↗