Effects of dietary antibiotics on intestinal microflora in broiler chickens

Changes were examined in the intestinal microflora in broiler chickens fed a diet containing antibiotics to obtain fundamental information on the mechanisms of beneficial effect of the antibiotics upon livestock production. Three antibiotics (colistin, bacitracin, and enramycin) were employed as feed additives. Experiments were conducted with broiler chickens in two ways. In one way dietary antibiotics were fed continually at levels approved for use as feed additives for a long term. In the other they were fed the same antibiotics for a short term. Significant changes in microflora were observed mainly in such bacterial groups as aerobic bacteria and Lactobacillus. In the long term administration, three possible modes of variance in the bacterial flora were postulated: Changes directly related to the antibacterial spectrum of antibiotics. Antagonistic changes related to an ecological balance in the bacterial flora. Changes in quantitative balance of bacteria constituting each bacterial group. The change in the intestinal microflora during administration of the antibiotic diet was expressed as a complex form of these transition modes. In the short term administration, it was demonstrated that the effect of the antibiotic diet lingered even 7 days after administration. This suggests that antibiotics used as feed additives may possibly affect the stability of the intestinal microflora.     

Effects of sodium butyrate and salinomycin upon intestinal microbiota, mucosal morphology and performance of broiler chickens

The effect of dietary sodium butyrate (SB) or salinomycin (SAL) or both additives on performance, small intestinal morphology and microbial ecology of broiler chickens was studied. A growth trial was conducted with 96 Ross 308 female broilers from 1 to 30 days of age. Four treatment groups were fed with a non-supplemented control diet or three experimental diets supplemented with i) 300 mg SB (Adimix 30 coated) per kg, ii) 60 mg SAL (Sacox) per kg or iii) both additives in combination. Feed intake and body-weight gain decreased and gain-to-feed ratio increased due to SAL supplementation, while addition of SB did not affect performance in comparison with the control diet but positively affected feed intake and body-weight gain in comparison with birds fed the SAL-supplemented diet. Villus height in jejunum decreased, while crypt depth increased due to SAL supplementation. Addition of SB increased crypt depth in jejunum. No significant effect of either additive was observed in ileum morphology. Total short-chain organic acids concentration in ileal digesta decreased with SAL supplementation, mainly due to lower lactic acid concentration, but no effects were observed in the caeca. The SAL supplementation was accompanied by a pH increase in ileum and a pH decrease in caecum. No significant effect of SB addition was observed for these parameters. Total bacterial numbers and Lactobacillus [lactic acid bacteria (LAB)] counts in ileal and caecal contents were lower in birds fed with SAL-supplemented diet in comparison with birds fed with control or SB diet. DNA fingerprints revealed SAL supplementation to affect the microbial population by suppressing dominating LAB, potentially L. aviarius. The presented results show that dietary SAL, supplemented alone or in combination with SB, suppressed the microbial activity and altered the microbial community structure mainly in ileum. SAL alone negatively affected feed intake and body-weight gain; however, the effect was ameliorated by SB supplementation.     

Effects of sodium butyrate and salinomycin upon intestinal microbiota,mucosal morphology and performance of broiler chickens

The effect of dietary sodium butyrate (SB) or salinomycin (SAL) or both additives on performance, small intestinal morphology and microbial ecology of broiler chickens was studied. A growth trial was conducted with 96 Ross 308 female broilers from 1 to 30 days of age. Four treatment groups were fed with a non-supplemented control diet or three experimental diets supplemented with i) 300 mg SB (Adimix 30 coated) per kg, ii) 60 mg SAL (Sacox) per kg or iii) both additives in combination. Feed intake and body-weight gain decreased and gain-to-feed ratio increased due to SAL supplementation, while addition of SB did not affect performance in comparison with the control diet but positively affected feed intake and body-weight gain in comparison with birds fed the SAL-supplemented diet. Villus height in jejunum decreased, while crypt depth increased due to SAL supplementation. Addition of SB increased crypt depth in jejunum. No significant effect of either additive was observed in ileum morphology. Total short-chain organic acids concentration in ileal digesta decreased with SAL supplementation, mainly due to lower lactic acid concentration, but no effects were observed in the caeca. The SAL supplementation was accompanied by a pH increase in ileum and a pH decrease in caecum. No significant effect of SB addition was observed for these parameters. Total bacterial numbers and Lactobacillus [lactic acid bacteria (LAB)] counts in ileal and caecal contents were lower in birds fed with SAL-supplemented diet in comparison with birds fed with control or SB diet. DNA fingerprints revealed SAL supplementation to affect the microbial population by suppressing dominating LAB, potentially L. aviarius. The presented results show that dietary SAL, supplemented alone or in combination with SB, suppressed the microbial activity and altered the microbial community structure mainly in ileum. SAL alone negatively affected feed intake and body-weight gain; however, the effect was ameliorated by SB supplementation.     

金针菇菇脚对肉鸡肠道菌群的影响

目的 研究金针菇菇脚（Flammulina velutipes stembase,FVS）对肉鸡生长性能、肠道发育及肠道菌群的影响。方法 选取4 550只1日龄爱拔益加（AA）肉鸡,随机分为5组：空白组（基础日粮）、2% FVS组（基础日粮+2% FVS）、4% FVS组（基础日粮+4% FVS）、6% FVS组（基础日粮+6% FVS）和6% FVS阶段组（基础日粮+试验第1、3、5周饲喂含有6%FVS的日粮,于试验2、4、6周饲喂基础日粮）。试验为期42 d。结果 FVS组肉鸡平均日增重量均显著高于空白组（P<0.05）。6% FVS组和6% FVS阶段组肉鸡平均日采食量显著高于空白组（P<0.05）。FVS组肉鸡料重比均显著低于空白组（P<0.05）。与空白组相比,日龄21 d时,2% FVS组肉鸡回肠和盲肠重量均显著增加（P<0.05）;4% FVS组十二指肠长度和盲肠重量均显著增加（P<0.05）;6% FVS阶段组十二指肠长度及重量、空肠长度及重量和回肠重量均显著增加（P<0.05）。与空白组相比,日龄42 d时,2% FVS组肉鸡十二指肠长度和重量及盲肠重量均显著增加（P<0.05）;4% FVS组十二指肠长度和重量、回肠长度和盲肠长度及重量均显著增加（P<0.05）;6% FVS组十二指肠长度和盲肠长度及重量均显著增加（P<0.05）;6% FVS阶段组十二指肠重量、空肠重量和回肠长度及重量均显著增加（P<0.05）。与空白组相比,日龄21 d时6% FVS阶段组肉鸡盲肠菌群DNA条带数显著增加;日龄42 d时FVS组肉鸡盲肠菌群DNA条带数均显著增加。日龄21 d和42 d时肉鸡盲肠菌群共性条带中均含有扭链胃球菌（Ruminococcus torques）,而FVS组含有大量的产酸菌。结论 日粮中添加FVS可提高肉鸡生长能力、促进肠道发育、增加肠道菌群多样性。     

Acidified litter benefits the intestinal flora balance of broiler chickens

The alterations in the balance of the normal intestinal bacterial flora of chickens exposed to acidified wood-derived litter were analyzed and compared to those of a control group exposed to nonacidified litter. A total of 1,728 broilers were divided into two groups, with six replicates in each. One group was exposed to dry wood-derived litter, and the other was exposed to dry wood-derived litter sprayed with a mixture of sodium lignosulfonate, formic acid, and propionic acid. At five different times, five chickens from each pen were killed and the intestinal contents from ileum and caeca were collected. The samples were diluted and plated onto selective media to identify coliforms, Lactobacillus spp., Clostridium perfringens, and Enterococcus spp. Covariance analysis of bacterial counts showed significantly lower counts for C. perfringens in the caeca and the ileum and for Enterococcus spp. and Lactobacillus spp. in the ileum in chickens exposed to the acidified litter. Lactobacillus spp. showed significantly higher counts in the caeca in chickens exposed to acidified litter. There was no difference between the two litters with regard to coliforms in the ileum and the caeca or to Enterococcus spp. in the caeca. The study shows that exposing the chickens to acidified litter lowers the intestinal bacterial number, especially in the ileum, without negative consequences for the chicken's health or performance. Of special interest are the lower counts of C. perfringens and Enterococcus spp. that might reduce the risk of developing clinical or subclinical necrotic enteritis and growth depression.     

The effect of microbial challenge on the intestinal proteome of broiler chickens

Background

In poultry production intestinal health and function is paramount to achieving efficient feed utilisation and growth. Uncovering the localised molecular mechanisms that occur during the early and important periods of growth that allow birds to grow optimally is important for this species. The exposure of young chicks to used litter from older flocks, containing mixed microbial populations, is a widely utilised model in poultry research. It rarely causes mortality but effects an immunogenic stimulation sufficient enough to cause reduced and uneven growth that is reflective of a challenging growing environment.

Methods

A mixed microbial challenge was delivered as used litter containing Campylobacter jejuni and coccidial oocysts to 120 male Ross 308 broiler chicks, randomly divided into two groups: control and challenged. On day 12, 15, 18 and 22 (pre- and 3, 6 and 10 days post-addition of the used litter) the proximal jejunum was recovered from 6 replicates per group and differentially abundant proteins identified between groups and over time using 2D DiGE.

Results

The abundance of cytoskeletal proteins of the chicken small intestinal proteome, particularly actin and actin associated proteins, increased over time in both challenged and control birds. Villin-1, an actin associated anti-apoptotic protein, was reduced in abundance in the challenged birds indicating that many of the changes in cytoskeletal protein abundance in the challenged birds were as a result of an increased rate of apoptosis. A number of heat shock proteins decreased in abundance over time in the intestine and this was more pronounced in the challenged birds.

Conclusions

The small intestinal proteome sampled from 12 to 22 days of age showed considerable developmental change, comparable to other species indicating that many of the changes in protein abundance in the small intestine are conserved among vertebrates. Identifying and distinguishing the changes in proteins abundance and molecular pathways that occur as a result of normal growth from those that occur as a result of a challenging microbial environment is important in this major food producing animal.

     

Role of volatile fatty acids in development of the cecal microflora in broiler chickens during growth

It is known that volatile fatty acids can inhibit growth of species of the family Enterobacteriaceae in vitro. However, whether these volatile fatty acids affect bacterial populations in the ceca of chickens is unknown. Therefore, a study was conducted to investigate if changes in volatile fatty acids in ceca of broiler chickens during growth affect bacterial populations. Results showed that members of the Enterobacteriaceae and enterococci are present in large numbers in 3-day-old broilers and start to decrease when broilers grow older. Lactobacilli are present in large numbers as well in 3-day-old broilers, but they remain stable during the growth of broilers. Acetate, butyrate, and propionate increase from undetectable levels in 1-day-old broilers to high concentrations in 15-day-old broilers, after which they stabilize. Significant negative correlations could be calculated between numbers of Enterobacteriaceae and concentrations of undissociated acetate, propionate, and butyrate. Furthermore, pure cultures of Enterobacteriaceae isolated from the ceca were grown in the presence of volatile fatty acids. Growth rates and maximal optical density decreased when these strains grew in the presence of increasing volatile fatty acid concentrations. It is concluded that volatile fatty acids are responsible for the reduction in numbers of Enterobacteriaceae in the ceca of broiler chickens during growth.     

Effect of cereal type and plant extract addition on the growth performance,intestinal morphology,caecal microflora,and gut barriers gene expression of broiler chickens

Feeding broiler chickens on diets based on cereal grains of high non-starch polysaccharides content such as wheat and barley can negatively impact their performance and gut health. Plant extracts can be used as a potential tool to alleviate these negative effects. The present study assessed the effects of dietary cereal type and the inclusion of a plant extract blend (PEB) on the growth performance, intestinal histomorphology, caecal microflora, and gene expression of selected biomarkers for gut integrity in broiler chickens in a 42-d experiment. Ross-308 male broilers were assigned into different dietary treatments and fed on two cereal types (corn- vs. wheat/barley-based) with/without added graded concentrations of a PEB (0, 250, 500, 1000, and 2000 mg/kg diet). There were no significant differences in the growth performance parameters, intestinal histomorphology, and caecal microflora due to the impact of dietary cereal type. However, lactobacilli count in the caecal microflora was increased in the group fed on a corn-based diet. The PEB supplementation especially at a level of 500 to 1000 mg/kg diet significantly increased the average BW and decreased the feed conversion ratio. It also increased the villi length of duodenum, jejunum, and ileum, decreased the duodenal crypt depth, and increased the villi length to crypt depth ratio in the duodenum, jejunum and ileum. Supplementation of the PEB decreased the total bacterial and coliform count and increased the lactobacilli count in a linear pattern. Gene expression of Occludin and Junction Adhesion Molecule was significantly increased in the PEB supplemented diets, whereby no influence was observed on mucin expression. In conclusion, supplementation of a PEB at levels of 500–1000 mg/kg can be used as a tool to improve broiler performance and gut health.     

Indigestible carbohydrates alter the intestinal microbiota but do not influence the performance of broiler chickens

Aims:  Prebiotics are a potential alternative to in-feed antimicrobials to improve performance of chickens. We investigated the effects of mannanoligosaccharide (MOS) and fructooligosaccharide (FOS) on growth, performance and the intestinal microbiota.
Methods and Results:  Cobb 500 birds were fed either: Control, starter diet without antimicrobials; ZnB, Control + 50 ppm zinc bacitracin; MOS, Control + 5 g kg⁻¹ MOS; or FOS, Control + 5 g kg⁻¹ FOS. An energy metabolism study was conducted and intestinal microbial communities assessed by T-RFLP and Lac PCR-DGGE. Diet did not influence performance. Ileal microbial communities were significantly different in ZnB-fed birds compared to all diets, and FOS-fed chickens compared to Control. MOS-fed chickens had a different caecal profile to ZnB and FOS-fed birds. Consensus Lac PCR-DGGE profiles indicated Lactobacillus communities clustered according to diet with Lactobacillus johnsonii characteristic of ZnB diet. Control and MOS-fed chickens displayed significantly different jejunal Lactobacillus profiles to each other whilst ileal profiles were different between MOS and FOS-fed birds.
Conclusion:  Prebiotics influenced the intestinal microbiota, but did not affect performance.
Significance and Impact of the Study:  In light of pressure for in-feed antimicrobial withdrawal, the impact of alternative compounds on the intestinal microbiota and bird performance is critical to the poultry industry.     

Effects of inulin and di-d-fructose dianhydride-enriched caramels on intestinal microbiota composition and performance of broiler chickens

In vitro and in vivo experiments were designed to evaluate the effectiveness of laboratory-made di-d-fructose dianhydride (DFA)-enriched caramels. The DFA-enriched caramels were obtained from d-fructose (FC), d-fructose and sucrose (FSC), or d-fructose and β-cyclodextrin (FCDC). In the in vitro experiment, raftilose and all caramels increased (P<0.05) l-lactate concentration and decreased (P<0.05) pH. Total short-chain fatty acid concentration was higher (P<0.05) than controls in tubes containing raftilose, FSC, FCDC and commercial sucrose caramel (CSC). Raftilose, and all caramels tested except FSC and FC (1%), increased (P<0.01) lactobacilli log₁₀ number of copies compared with the non-additive control. FSC, FCDC and CSC increased (P<0.01) the bifidobacteria number of copies as compared with controls. All additives, except FCDC, decreased (P<0.01) Clostridium coccoides/Eubacterium rectale log number of copies. Compared with controls, raftilose, FC and CSC led to lower (P<0.01) Escherichia–Shigella and enterobacteria. For the in vivo experiment, a total of 144 male 1-day-old broiler chickens of the Cobb strain were randomly assigned to one of the three dietary treatments for 21 days. Dietary treatments were control (commercial diet with no additive), inulin (20 g inulin/kg diet) and FC (20 g FC/kg diet). Final BW of birds fed FC diet was higher (P<0.01) than controls or inulin-fed birds, although feed: gain values were not different. Feed intake of chickens fed FC was higher (P<0.01) than that of inulin-fed birds but not statistically different from controls. Crop pH values were lower (P<0.01) in birds fed FC diet as compared with control diet, with inulin-fed chickens showing values not different from control- or FC-fed birds. Lower (P<0.05) lactobacilli number of copies was determined in the crop, ileum and caeca of birds fed the inulin diet compared with the control diet. Inulin supplementation also resulted in lower (P<0.05) C. coccoides/E. rectale, bacteroides and total bacteria in caecal contents. Addition of FC to broiler diets gave place to lower (P<0.05) enterobacteria and Escherichia–Shigella in crop and caecal contents compared with controls. The bacteroides number of copies increased (P<0.05) as compared with controls in the ileum, but decreased (P<0.05) in the caeca of chickens fed the FC diet. Energy, ADF, NDF and non-starch polysaccharides faecal digestibilities were greater (P<0.05) than controls in chickens fed diets containing inulin or FC. Fat digestibility was higher (P<0.05) in FC-fed birds compared with controls or inulin-fed chickens. In conclusion, DFA-enriched caramels tested here, particularly FC, may represent a type of new additives useful in poultry production.     

Impact of antimicrobial usage on antimicrobial resistance in commensal Escherichia coli strains colonizing broiler chickens

Escherichia coli strains isolated from commercial broilers and an experimental flock of chickens were screened to determine phenotypic expression of antimicrobial resistance and carriage of drug resistance determinants. The goal of this study was to investigate the influence of oxytetracycline, sarafloxacin, and enrofloxacin administration on the distribution of resistance determinants and strain types among intestinal commensal E. coli strains isolated from broiler chickens. We detected a high prevalence of resistance to drugs such as tetracycline (36 to 97%), sulfonamides (50 to 100%), and streptomycin (53 to 100%) in E. coli isolates from treated and untreated flocks. These isolates also had a high prevalence of class 1 integron carriage, and most of them possessed the streptomycin resistance cassette, aadA1. In order to investigate the contribution of E. coli strain distribution to the prevalence of antimicrobial resistance and the resistance determinants, isolates from each flock were DNA fingerprinted by enterobacterial repetitive intergenic consensus sequence (ERIC) PCR. Although very diverse E. coli strain types were detected, four ERIC strain types were present on all of the commercial broiler farms, and two of the strains were also found in the experimental flocks. Each E. coli strain consisted of both susceptible and antimicrobial agent-resistant isolates. In some instances, isolates of the same E. coli strain expressed the same drug resistance patterns although they harbored different tet determinants or streptomycin resistance genes. Therefore, drug resistance patterns could not be explained solely by strain prevalence, indicating that mobile elements contributed significantly to the prevalence of resistance.     

Spatial and temporal variation of the intestinal bacterial community in commercially raised broiler chickens during growth

The objective of this study was to determine whether host, compartment, or environmental specific factors play an important role in the establishment of the intestinal microflora in broiler chickens during growth. This objective was addressed using a 16S rDNA approach. PCR-amplicons from the V6 to V8 regions of the 16S rDNA of intestinal samples were separated by denaturing gradient gel electrophoresis (DGGE). The number of bands in all intestinal compartments increased when broilers grew older, indicating that the dominant bacterial community becomes more complex when chickens age. Each chicken had a unique banding pattern for all locations in the intestinal tract, irrespective of the age of chickens. This suggests that host-related factors affect the establishment of the dominant bacterial community. Banding patterns of intestinal compartments within one chicken were different from each other for broilers older than 4 days, except for both ceca which were highly similar. In 4-day-old broilers, banding patterns from crop, duodenum, and ileum were very similar. We conclude that (unknown) host specific factors play an important role in the development of the intestinal bacterial community in each broiler chicken. Furthermore, compartment-specific factors play an important role in the bacterial development of each intestinal compartment within one chicken.     

Effect of raw and fermented grape seed on growth performance,antioxidant capacity,and cecal microflora in broiler chickens

Grape seed (GS) is a by-product of the fruit juice and wine industry with the potential to be an alternative to synthetic antioxidants due to its antioxidant activity. Agro-industrial residues can be converted to more effective products by solid-state fermentation. The objective of the study was to investigate the effects of GS and fermented grape seed (FGS) on the growth performance, antioxidant capacity, and cecal microflora in broiler chickens. A total of 128 female broilers were randomly allocated into four treatment groups, each consisting of four replicates of eight birds. Throughout the 42-day feeding period, the birds were fed with soybean-corn based diet (CON), 0.15 g/kg synthetic antioxidant (butylated hydroxyanisole) supplemented diet (AO), 5 g/kg GS supplemented diet (GS), and 5 g/kg FGS supplemented diet (FGS). Dietary GS, FGS, and AO supplementation increased the BW (P < 0.05) and average daily weight gain (ADG, P < 0.05) compared with the CON group in the overall period of 42 days. Dietary FGS also increased the ADG (P < 0.05) in the period of 22–42 days compared with the control group. The pH of the breast meat of the chickens fed GS was higher (P < 0.01) than CON and FGS groups. Dietary FGS and AO decreased the b* value (P < 0.01) of breast meat compared with the CON group. Grape seed had the highest serum glutathione peroxidase (P < 0.05) and catalase (CAT, P < 0.01) levels among the treatment groups. The FGS also increased serum CAT level (P < 0.01) compared with the AO group. Moreover, dietary FGS supplementation increased Lactobacillus spp. (P < 0.05) in the cecum compared with the other treatment groups and decreased Staphylococcus aureus (P < 0.05) compared with the CON and AO groups. The present findings indicate that GS and FGS can be used in broiler diets as alternatives to synthetic antioxidants.     

Minimum inhibitory concentrations of intestinal Escherichia coli from broiler chickens after oral administration of apramycin

This study describes the influence of apramycin on minimum inhibitory concentrations (MIC) of intestinal Escherichia coli in young broiler chickens, after oral administration of the antibiotic at a dosage equivalent to a prophylactic course of treatment for 10 d. The bacteria were isolated from cloacal swabs and caecal contents. MICs were determined by agar dilution procedures. MIC of apramycin for the investigated strains ranged from 1 μg ml^-1 to 16 μg ml^-1. Strains obtained from undosed birds mainly had MIC values of 1 μg ml^-1. MIC values of 8 μg ml^-1 or more were recorded only among isolates obtained from chickens which had received apramycin. Administration of apramycin resulted in a slight but statistically significant increase in the average MIC. Statistically higher average MICs were recorded among isolates from cloacal swabs 10 d after withdrawal until the end of the experiment. For strains from caecal contents, this was demonstrated only on one sampling occasion, 15 d after withdrawal.     

Inheritance of fertility in broiler chickens

Background

The fertility of a chicken''s egg is a trait which depends on both the hen that lays the egg and on her mate. It is also known that fertility of an individual changes over the laying period.

Methods

Longitudinal models including both random genetic and permanent environmental effects of both the female and her male mate were used to model the proportion of fertile eggs in a pedigree broiler population over the ages 29-54 weeks.

Results

Both the male and the female contribute to variation in fertility. Estimates of heritability of weekly records were typically 7% for female and 10% for male contributions to fertility. Repeatability estimates ranged from 24 to 33%, respectively. The estimated genetic variance remained almost constant for both sexes over the laying period and the genetic correlations between different ages were close to 1.0. The permanent environment components increased substantially towards the end of the analyzed period, and correlations between permanent environment effects at different ages declined with increasing age difference The heritability of mean fertility over the whole laying period was estimated at 13% for females and 17% for males. A small positive correlation between genetic effects for male and female fertility was found.

Conclusion

Opportunities to improve fertility in broiler stocks by selection on both sexes exist and should have an impact throughout the laying period.     

Effects of density on perching behavior of broiler chickens

Exercise through perching has been suggested as a way to reduce the impact of leg problems in broiler chickens. It is possible that higher stocking densities may motivate birds to perch more, and perhaps reduce some of the detrimental effects to broiler health seen with high stocking densities. The goal of this research was to investigate the effects of density (10, 15, and 20birds/m(2)) and perch design on the frequency of perch utilization. Mixed sex broilers were assigned to 36 pens in a four perch treatmentxthree density factorial with three replications for each treatment/density combination in a randomized complete block design. Pens were assigned to one of four perch treatments: control (no perches), horizontal (three horizontal perches), angled (three 10 degrees angled perches), or mixed angle (one horizontal, one 10 degrees angled, and one 20 degrees angled perch), at each of the three densities. The results show that although the frequency of perch use was low, (2.6%+/-0.15), significant preferences were clear. Perching frequency was significantly higher when birds were raised at stocking densities of 15 (2.88+/-0.303) or 20birds/m(2) (2.76+/-0.211), as opposed to 10birds/m(2) (2.10+/-0.209) as indicated by a marginally significant main effect (p<0.0597), and a significant linear effect (P<0.0427). The birds used the horizontal perches the most, followed by 10 degrees angled and mixed angle perch treatments with decreasing frequency. An effect of age was found, in that perching increased during the first 4 weeks, and dropped off significantly at the end of the rearing period for all perch treatments. In general, perches closest to the hallway or outside of the house were used more than perches in the middle of the pen. Preferential use of the highest section of the 10 degrees angled perches was also found. The applied perch treatments or densities did not significantly affect final body weight or feed conversion. Percent mortality due to heat stress showed a significant increase with density, while mortality not caused by heat stress did not change with perch treatment or density.     

Diversity of insect intestinal microflora

The influence of geographic location, season, age, and part of the digestive tract on bacterial diversity was evaluated on intestinal microflora of honeybees, wasps, and cockroaches using DGGE analysis. PCR-DGGE analyses with universal bacterial primers targeting 200-bp region of the 16S rDNA gene afforded the profile of complex bacterial DNA; specific primers were used to determine the profile of bifidobacteria whose concentration in digestive tract was determined by real-time PCR. Selected PCR products were identified by sequencing. The microflora of the bees exhibited little variations among the hives from distant locations. Their bifidobacterial population formed 2.8-8.4 % of total bacteria and was very homogeneous. The total gut microflora of wasps was also homogeneous, only two samples being affected by the season or the location; on the other hand, wasp bifidobacterial population was very heterogeneous. Cockroaches showed the highest variations in microflora composition, the age and diet being the ultimate factors; bifidobacteria counts also varied among tested individuals (0.1-34.1 % of total bacteria). Our results suggest that nutrition habits are the strongest factor affecting the insect microflora, giving higher variations to omnivorous species.