Phenotypic and genotypic characterization of Enterococcus spp. from yolk sac infections in broiler chicks with a focus on virulence factors.

Bacterial infections of yolk sacs contribute to increased mortality of chicks, chronic infections during their rearing, or increased selection in the flock, which in turn leads to high economic losses in poultry production worldwide. The aim of this study was a phenotypic and genotypic characterization of enterococci isolated from yolk sac infections (YSI) of broiler chickens from Poland and the Netherlands. Biochemical, matrix-assisted laser desorption/ionization (MALDI)-time-of-flight (TOF) MS, and rpoA gene sequencing identification was performed. Moreover, phenotypic and genotypic characterization of virulence factors and analysis of the clonal relationship of isolates by MALDI-TOF MS and enterobacterial repetitive intergenic consensus-polymerase chain reaction (ERIC-PCR) were performed. The biochemical test identified 70 isolates as Enterococcus faecalis and 6 as Enterococcus mundtii. The results of MALDI-TOF MS were 100% concordant with those obtained by rpoA gene sequencing, and all 76 isolates were identified as E. faecalis. Differences were noted in the ß-glucuronidase, ß-glucosidase, α-galactosidase, phosphatase, melibiose, lactose, and raffinose tests that is going about the results of biochemical identification. None of the isolates were beta-hemolytic on blood agar in aerobic conditions, but all but one were gelatinase positive. Among biofilm-forming isolates (30/76; 39.5%), as many as 66.7% (20/30) were Polish E. faecalis strains. Most of the isolates carried virulence genes, that is gelE, ace, asa1, efaAfs, fsrA, fsrB, fsrC, cob, cpd, and ccf, but none had the hyl gene. Some isolates harbored cyl operon genes. One Polish strain (ST16) had all of the tested cyl genes and the esp gene, considered clinically important, and showed the highest biofilm-forming ability. Nearly 50% of the isolates showed close genetic relatedness in ERIC typing. In contrast with MALDI-TOF MS cluster analysis, ERIC-PCR results did not show a relationship with the origin of the strains. Using MALDI-TOF MS, 7 peaks were found in Polish and Dutch isolates, which may type them as species-specific biomarkers in E. faecalis from YSI.

Avian antimicrobial peptides: in vitro and in ovo characterization and protection from early chick mortality caused by yolk sac infection.

Increasing antibiotic resistance is a matter of grave concern for consumers, public health authorities, farmers, and researchers. Antimicrobial peptides (AMPs) are emerging as novel and effective non-antibiotic tools to combat infectious diseases in poultry. In this study, we evaluated six avian AMPs including 2 truncated cathelicidins, [CATH-1(6-26) and CATH-2(1-15)], and 4 avian ß-defensins (ABD1, 2, 6 and 9) for their bactericidal and immunomodulatory activities. Our findings have shown CATH-1(6-26) and ABD1 being the two most potent avian AMPs effective against Gram-positive and Gram-negative bacteria investigated in these studies. Moreover, CATH-1(6-26) inhibited LPS-induced NO production and exhibited dose-dependent cytotoxicity to HD11 cells. While, ABD1 blocked LPS-induced IL-1ß gene induction and was non-toxic to HD11 cells. Importantly, in ovo administration of these AMPs demonstrated that ABD1 can offer significant protection from early chick mortality (44% less mortality in ABD1 treated group versus the control group) due to the experimental yolk sac infection caused by avian pathogenic Escherichia coli. Our data suggest that in ovo administration of ABD1 has immunomodulatory and anti-infection activity comparable with CpG ODN. Thus, ABD1 can be a significant addition to potential alternatives to antibiotics for the control of bacterial infections in young chicks.

Post hatch recovery of a probiotic Enterococcus faecium strain in the yolk sac and intestinal tract of broiler chickens after in ovo injection.

Concerns about antibiotic-resistant bacteria and their presence in animal products grow and thus alternatives to use of antibiotics in animal production are being investigated. Probiotics have gained increased focus due to improvements in performance, immune health and pathogen reduction when provided to poultry through feed. These traits may be further improved if probiotics can be provided to the embryo before hatch, before meeting environmental pathogens. The objective was to determine the faith of a probiotic Enterococcus faecium (M74) strain in the yolk sac and intestinal tract of broiler chickens after injection into hatching eggs. E. faecium M74 (1.4 × 107 CFU/egg) was applied in ovo at day 18 of incubation. From 1- and 7-day-old chickens, 20 samples from yolk sac, caecal tonsils and rest of the intestinal tract were subjected to CFU counting. Isolates from a sample subset were typed by pulsed-field gel electrophoresis (PFGE). Enterococci were found in varying numbers: 1.0 × 104-2.2 × 1010 CFU/g. The prevalence of M74 PFGE profiles was high in 1-day-old (88%) and 7-day-old chickens (67%). This demonstrates that the embryos ingested M74 before hatching, that M74 is viable for intestinal colonization through in ovo administration, and that the strain multiplies in the chickens gastrointestinal tract post hatching.

An evaluation of alternative methods for sanitizing hatching eggs.

Different sanitization methods were evaluated as alternatives to formaldehyde fumigation for the reduction of eggshell and yolk sac microbiological counts, improvement of eggshell quality, incubation parameters, and day-old chick quality. A total of 10,080 hatching eggs were collected from a 70-wk-old commercial broiler breeder flock and distributed in a completely randomized block design with seven treatments: fumigation with paraformaldehyde (5.03 g/m3/30 min), fumigation with ozone (5-15 ppm/30 min), ultraviolet light-C irradiation (8.09 mW/cm2; 120 s; UV-C), hydrogen peroxide spraying (3%; 0.69 mL/egg), peracetic acid spraying (0.3%; 0.69 mL/egg; PAA), water spraying (0.69 mL/egg; water control), and without disinfection (dry control-DC). Spraying eggs with PAA and UV-C significantly reduced aerobic bacteria plate counts compared to the DC group. In addition, eggs disinfected with PAA had lower Enterobacteriaceae counts than the DC and water control groups. Eggshell quality, incubation parameters, and microbiological counts for yolk sac did not differ (P > 0.05) among treatments. This study demonstrated the potential for the application of PAA and UV-C for eggshell disinfection instead of formaldehyde; however, an electronic microscopic evaluation of the eggshell is necessary to determine if these methods cause any damage to the cuticle.

[Can Chlamydia trachomatis human biovars cause abortion in cattle? An immunohistochemical study on a new host-pathogen relationship]. / Chlamydia trachomatis'in insan biyotipleri sigirlarda düsük etkeni olabilir mi? Yeni bir konak-patojen iliskisini arastiran immünohistokimyasal çalisma.

Chlamydiae, which are obligate intracellular bacterial pathogens, have been radiated from single-celled eukaryotes into multi-celled hosts during their evolution. Chlamydia trachomatis one of the important species in this group, is classified into three biovars as a result of their evolution. Two of those biovars, Trachoma and LGV, are pathogens only in humans. Initially, the presence of a high specificity between the host and chlamydiae has been recognized and this relation has been considered as an adaptation mechanism. However, some studies have indicated that chlamydiae can also grow in laboratory animals, yolk sacs of embryonated eggs and in vitro cell cultures. The aim of this study was to investigate if C. trachomatis human specific biovars are possible infectious agents in the aborted bovine fetuses. Ninety aborted bovine fetuses were included in the study, and the bacteria which could be the causative agents for abortion were searched by conventional microbiological methods. Twenty-three (25.6%) abortion materials which have yielded negative results with these methods for the presence of bacterial agents other than chlamydiae, were further evaluated in terms of the presence of C. trachomatis. For this purpose the samples were inoculated into the yolk sac of embryonated eggs and the slides prepared from the yolk sac membranes of embryons died after 24 hours of inoculation, were examined for the presence of inclusion bodies by staining with Giemsa method. The presence of C. trachomatis specific antigens and glycogen inclusions in those 23 samples were also investigated by immunohistochemical and Lugol's iodine staining methods, in the fetal tissue samples which were embedded in paraffin. Immunohistochemical method was performed with immunoperoxidase staining by the use of specific antibodies against C. trachomatis major outer membrane proteins. As a result, 5 (21.7%) of the 23 samples were found positive for C. trachomatis with three of the methods (Giemsa, immunoperoxidase and lugol stainings). Although the data of our study have supported that chlamydiae can adapt to new host species other than humans, further advanced studies are needed on this subject. Our results have also emphasized that novel routes of transmission should be considered for C. trachomatis infections.

Fumigating broiler hatching eggs with lysozyme product (Inovapure) to reduce eggshell microbial load.

Experiments were conducted to evaluate the effectiveness of a lysozyme product (InovapureTM) (LP) against E. coli penetrating eggshells. In the first microbiological experiment, 60 agar-filled eggs were inoculated with E. coli suspension, then fumigated with distilled water, 1.5% or 3.0% LP or a quaternary ammonium product (QA) at 0.125% for 10 min. In the second microbiological experiment, another 60 agar-filled eggs were fumigated with the same sanitizer treatments first, then inoculated with the E. coli suspension. Eggshells were candled and visual colonies were counted after 48 h incubation. An animal experiment was conducted to evaluate LP applied to the surface of 2080 broiler hatching eggs on hatching and growth performance. Hatching eggs were submerged in an E. coli suspension. After drip drying, eggs were randomly divided into four fumigation treatments, each with four subsets of 150 eggs. Fumigation treatments were the same as in the microbiological experiments. Eggs were incubated in 8 incubators (2 replicate incubators per treatment) and the broilers were grown to 33 d of age. In the microbiological experiments, inoculated eggs fumigated with 3.0% LP and 0.125% QA reduced (P < 0.05) the total amount of E. coli to 11 cfu/egg and 10 cfu/egg, respectively. When eggs were sanitized prior to inoculation, 3.0% LP demonstrated (P < 0.05) ongoing bactericidal action to prevent E. coli penetration. No differences in hatchability, fertility rate or egg weight loss percent were found among sanitation treatments. At hatch, body weight or the ratio of yolk sac weight to yolk-free body weight were not affected by the sanitation treatments. However, the application of sanitizers decreased (P < 0.05) the presence of E. coli in the yolk sac of newly hatched chicks. Feed consumption, body weight and feed conversion ratio were not affected by sanitation treatments. However, average daily body weight gain was lower (P < 0.05) following QA. Overall, 3.0% LP demonstrated acceptable activity against E. coli on eggshells, and provided ongoing bactericidal action to prevent E. coli penetration without negatively affecting growth performance.

Poly-ß-hydroxybutyrate administration during early life: effects on performance, immunity and microbial community of European sea bass yolk-sac larvae.

The reliable production of marine fish larvae is one of the major bottlenecks in aquaculture due to high mortalities mainly caused by infectious diseases. To evaluate if the compound poly-ß-hydroxybutyrate (PHB) might be a suitable immunoprophylactic measure in fish larviculture, its capacity to improve immunity and performance in European sea bass (Dicentrarchus labrax) yolk-sac larvae was explored. PHB was applied from mouth opening onwards to stimulate the developing larval immune system at the earliest possible point in time. Larval survival, growth, microbiota composition, gene expression profiles and disease resistance were assessed. PHB administration improved larval survival and, furthermore, altered the larva-associated microbiota composition. The bacterial challenge test using pathogenic Vibrio anguillarum revealed that the larval disease resistance was not influenced by PHB. The expression profiles of 26 genes involved e.g. in the immune response showed that PHB affected the expression of the antimicrobial peptides ferritin (fer) and dicentracin (dic), however, the response to PHB was inconsistent and weaker than previously demonstrated for sea bass post-larvae. Hence, the present study highlights the need for more research focusing on the immunostimulation of different early developmental stages for gaining a more comprehensive picture and advancing a sustainable production of high quality fry.

Presence of inoculated Campylobacter and Salmonella in unabsorbed yolks of male breeders raised as broilers.

Day-old male broiler breeder chicks were obtained from a commercial hatchery and raised as broilers. For Experiment 1, at 5 wk of age, the broilers were orally inoculated with a 10(6) cfu/ml of a characterized strain of Campylobacter jejuni and a cocktail (three naladixic acid-resistant strains) of Salmonella serovars. One week after inoculation, the birds were euthanatized and defeathered. The abdominal cavity was examined and any unabsorbed yolk material (and remaining yolk stalk) and ceca were aseptically removed for microbiological analyses. For each pooled sample (two birds per pool), an aerobic plate count (APC), an Enterobacteriaceae (ENT) count, and a test for the presence of Campylobacter and Salmonella was performed. For Experiment 2, at 5 wk of age, the broilers were orally inoculated with 10(5) cfu/ml of a characterized strain of Campylobacter jejuni. One week after inoculation, the birds (n = 20) were killed, defeathered, and the yolk stalk, attached yolk, or free-floating yolk and ceca were individually analyzed for presence of Campylobacter. For Experiment 1, the Salmonella-inoculated birds had 2/12 ceca and 0/12 unabsorbed yolk samples positive for Salmonella. The average yolk APC was log10 3.4 cfu/g and the average ENT was log10 1.9 cfu/g. For the Campylobacter-inoculated birds, 12/12 ceca and 9/12 unabsorbed yolk samples were positive for Campylobacter. The average yolk APC was log10 3.5 cfu/g and the average ENT was log10 3.1 cfu/g. For Experiment 2, the inoculated Campylobacter birds had 19/20 ceca, 5/20 free floating yolks, and 19/20 yolk stalks positive. In Experiment 1, the inoculated Campylobacter colonized the ceca in every instance and were present in 75% of the unabsorbed yolks. Alternatively, the inoculated Salmonella were not found in any of the unabsorbed yolks and only rarely in the ceca. In Experiment 2, the inoculated Campylobacter was found in very high numbers in the yolk and internal body samples. Determining to what extent these internal bodies and unabsorbed yolks play in bacterial colonization and contamination of the birds at processing has not been determined. The next step will be to determine the incidence of unabsorbed yolks and presence of Campylobacter and Salmonella in these bodies of commercial broilers at processing.

Omphalitis associated with Aspergillus fumigatus in poults.

Omphalitis associated with aspergillosis was diagnosed in four cases of commercial turkey poults ranging in age from 3 to 9 days old. In two cases, the mycotic agent present in the yolk sac was isolated and identified as Aspergillus fumigatus. In the other two cases, the fungi were identified as Aspergillus sp. on the basis of morphologic characteristics of the fungi in tissue sections. The fungi present were further confirmed to be of the genus Aspergillus by immunohistochemistry. Omphalitis by A. fumigatus infection has not been documented before.

Phenotypic characterization of ipaH+ Escherichia coli strains associated with yolk sac infection.

Seventy-six Escherichia coli serotypes possessing the ipaH gene typical of enteroinvasive E. coli (EIEC) strains were characterized. Biochemical identification of our strains shows positive reactions for lactose fermentation (100% of strains), lysine decarboxylase (98.7% of strains) and motility (67.1% of strains), properties that do not correspond with those described to the EIEC group. The serotypes agree with an initial classification. In this, some common O antigens identified among ipaH+ strains were O2 (n=20), OR (n=11) and non-determined O? (n=10). The O2:NM serotype was the most common. Sixty-six percent (n=50) of the ipaH+ E. coli strains were colicin producers, of them, 26 (34%) produced Col V and other colicins, 13 (17%) produced colicins other than Col V, and 11 (14.5%) produced Col V only. Trimethoprim/Sulfa (72%), ampicillin (64.5%), enrofloxacin (55.3%), and ciprofloxacin (47.4%) were the major antimicrobial resistance frequencies observed. Twenty-five different multiresistance patterns were observed, where sixty-six strains (86.8%) were included. A MIC test showed that most of the strains were sensitive to low gentamicin and kanamycin concentrations, whereas most of the strains were resistant to tetracycline. An invasiveness assay showed that the predominant alterations caused to HEp-2 cells were changes in shape and staining, and in most of the specimens, a partial monolayer detachment was also seen. Fifteen strains invaded more than 30% of the monolayer cells, causing the formation of intercellular bridges or filipoidal-like protrusions. The results suggest the existence of specific clone complexes derived from EIEC strains adapted to the avian host. To our knowledge, this is the first study that demonstrates the presence of extraintestinal invasive E. coli (ExIEC) strains.

Experimental infection of chicken embryos with recently described Brucella microti: Pathogenicity and pathological findings.

Brucellae are facultative intracellular pathogens causing disease in a wide range of domestic and wild animals as well as in humans. Brucella (B.) microti is a recently recognized species and was isolated from common voles (Microtus arvalis), red foxes and soil in Austria and the Czech Republic. Its pathogenicity for livestock and its zoonotic potential has not been confirmed yet. In the present study 25 SPF chicken embryos were inoculated at day 11 of age with 1.6×10(3) and 1.6×10(5)B. microti by yolk sac and allantoic sac routes. Re-isolation of B. microti indicated rapid multiplication of bacteria (up to 1.7×10(12)CFU). B. microti provoked marked gross lesions, i.e. hemorrhages and necroses. All inoculated embryos were dead (100% mortality) in between 2nd and 4th day post inoculation. The predominant histopathological lesion was necroses in liver, kidneys, lungs, spleen, gastrointestinal tract, spinal meninges, yolk sac and chorioallantoic membrane. Immunohistochemical examination showed the presence of Brucella antigen in nearly all of these organs, with infection being mainly restricted to non-epithelial cells or tissues. This study provides the first results on the multiplication and pathogenicity of the mouse pathogenic B. microti in chicken embryos. These data suggest that, even though chicken are not mammals, they could provide a useful tool for understanding the pathogenesis of B. microti associated disease.

Investigations on the pathogenicity of Listeria spp. by experimental infection of the chick embryo.

To investigate the pathogenicity of Listeria spp., chick embryos were infected by two methods. Very encouraging results were obtained with the inoculation of the chorioallantoic membrane (CAM) of 10 day old chick embryos: embryos inoculated with pathogenic strains (L. monocytogenes or L. ivanovii) died within 72 h, whereas those infected with apathogenic strains survived. This test could be a suitable method to replace the mouse pathogenicity test.

Abortion in Japanese cows caused by Chlamydia psittaci.

An outbreak of abortion in cows occurring in Niigata Prefecture was shown to be caused by Chlamydia psittaci. Elementary bodies characteristic of Chlamydia were found in the liver of aborted fetuses and C. psittaci antigen was demonstrated by indirect immunofluorescence. Chlamydia was isolated from the liver of aborted fetuses by the yolk sac inoculation of developing chick embryos and by the intraperitoneal inoculation of guinea pigs. Abortion occurred mostly in middle or late pregnancy. Aborted fetuses showed subcutaneous edema and gelatinous infiltration, enlarged liver and spleen, and dark red pleural and ascitic fluid. Focal necrosis was shown in the liver, spleen and lymph nodes. Serological findings and isolation of Chlamydia from fecal specimens indicated a wide dissemination of C. psittaci among cows in the area.

Salmonella typhimurium penetration through the eggshell of hatching eggs.

This study was undertaken to demonstrate penetration of Salmonella typhimurium through the eggshell of newly laid broiler hatching eggs. Eggs were challenged either by lightly spraying the bacteria over the blunt end of the egg or by contact with contaminated dry nest litter. Exposure time for both groups was 10 minutes; afterward, all eggs were disinfected and incubated 19 days under normal conditions. Chorioallantoic membranes and yolk sacs were cultured in brain-heart infusion broth on day 19 to demonstrate penetration. Isolation of the bacteria from chorioallantoic membranes and yolk sacs, respectively, were as follows: sprayed group: 100% and 83%; contact group: 59% and 29%. These results showed that although water enhanced S. typhimurium penetration, its presence on the eggshell is not essential for penetration to occur.

Chicken embryo propagation of type I avian adenoviruses.

Forty-two clone-purified, cell-culture-propagated type I avian adenoviruses (AAV) representing 11 serotypes and two intermediate strains were evaluated for virus replication (evidenced by embryo death and lesions) resulting from the inoculation of specific-pathogen-free chicken embryos via the chorioallantoic sac or yolk sac. Commonly observed embryonic changes were death, stunting and curling, hepatitis, splenomegaly, congestion and hemorrhage of body parts, and urate formation in the kidneys. Basophilic or eosinophilic intranuclear inclusion bodies characteristic of fowl adenoviruses were observed in hepatocytes. The magnitude and relative uniformity of intra- and interserotypic embryo mortality, gross lesions, and virus titers was greater in embryos inoculated via the yolk sac. This work identifies the yolk sac as a practical and sensitive chicken embryo inoculation route for poultry diagnosticians to employ. It is suggested that the yolk sac may be a reliable alternative to cell culture for the successful isolation of all type I avian adenoviruses.

Effect of hydrogen peroxide disinfection during incubation of chicken eggs on microbial levels and productivity.

Hatchery sanitation has a significant impact on chick quality. The proper use of disinfectants is essential. Aerosol bacterial counts, egg moisture loss, hatchability, chick quality, and broiler productivity were measured in eggs exposed to hydrogen peroxide fogging and compared with eggs not exposed to disinfectant during the incubation period. Hydrogen peroxide was also evaluated in the presence of a severe challenge with Staphylococcus aureus-contaminated eggs. A significant reduction was found in aerosol bacterial counts within the hatcher when incubators were fogged with 3% hydrogen peroxide when compared with water-fogged machines even in the face of high bacterial challenge. Eggs exposed to hydrogen peroxide lost a significantly greater amount of moisture during incubation, but hatchability was not affected. The use of hydrogen peroxide as a hatchery sanitizer did not affect broiler livability, body weight, or feed conversion but did reduce the incidence of retained yolk sacs in 42-day-old chickens.

Serotyping and virulence genes detection in Escherichia coli isolated from fertile and infertile eggs, dead-in-shell embryos, and chickens with yolk sac infection.

Escherichia coli is a common avian pathogen mainly associated with extraintestinal infections such as yolk sac infection (YSI). The aim of this study was to determine the serotypes and the presence of some virulence genes of E. coli strains isolated from different samples in a vertically integrated poultry operation in Mexico. Two hundred sixty-seven E. coli isolates from different samples were serotyped using rabbit serum against the 175 somatic (O) and 56 flagellar (H) antigens of the typing schema. Virulence genes were determined by colony blot hybridization, using DNA probes for st, eae, agg1, agg2, bfp, lt, cdt, slt, and ipaH diarrhea-associated virulence factors. The serogroup of 85% of the strains was determined; O19 (12%), 084 (9%), 08 (6%), and 078 (5%) were the most common. Using the complete antigenic formula (O and H), O19:NM (n = 31) was the serotype most frequently isolated from dead-in-shell embryos and in broilers that had died on the fourth, fifth, sixth, and seventh days after hatch. One hundred ten strains (41.2%) hybridized with one or more of the used probes. Of these, ipaH (72%), eae (30%), and cdt (27%) were the most common. Considering the origin of the respective isolates, 40% of the broiler farm strains were positive for at least one probe. Results show that some avian E. coli strains isolated in Mexico are included in avian pathogenic E. coli serotypes not previously reported, suggesting that they could be specific for this geographic area. The wide distribution of the ipaH gene among nonmotile strains suggests that this invasiveness trait could be important in YSI pathogenesis. On the other hand, some other genes could contribute to E. coli virulence during YSI.

The effect of inoculating Enterococcus faecalis into the yolk sac on chick quality and maternal antibody absorption.

Four hundred thirty-two 1-day-old specific-pathogen-free chicks were randomly divided into 36 groups of 12. All chicks were given 0.2 ml of Newcastle disease antiserum (hemagglutination-inhibition [HI] titer 1:5120) by injection into the yolk sac at hatch. Half of the groups received 0.2 ml of Enterococcus faecalis (4.0 x 10(8) colony-forming units/ml) by injection into the yolk sac at hatch (treatment). The remaining 18 groups received no bacteria (control). Two treatment groups and two control groups were weighed, bled, killed, and yolk sac weighed daily for the first 9 days of life. Feed was weighed at placement and at the end of the trial. Blood was tested for packed cell volume (PCV), total plasma protein, and Newcastle disease HI titer. No significant difference was observed between treatment and control groups for chick body weight, PCV, and feed consumption. Total plasma protein and retained yolk weight were significantly higher for treatment groups over control (P < 0.01 and P < 0.0001, respectively). Also, the geometric mean serum HI titer (log2) for Newcastle disease antibody was significantly higher in the control chicks vs. the treatment chicks (P < 0.01).

Effect of BioSentry 904 and ethylenediaminetetraacetic acid-tris disinfecting during incubation of chicken eggs on microbial levels and productivity of poultry.

Proper sanitation practices and the use of efficacious disinfectants in a hatchery have an effect on chick quality. Aerosol bacterial counts, egg moisture loss, hatchability, chick quality, and broiler productivity were evaluated when egg surfaces were contaminated by immersion of each egg into a broth medium containing a field isolate of Pseudomonas aeruginosa and incubated with exposure to one of three disinfectant treatments administered by fine spray: distilled water, BioSentry 904 (904), and a 1:1 ratio of 904 and ethylenediaminetetraacetic acid (EDTA)-Tris. The aerosol bacteria levels were statistically greater on day 21 of incubation in the group treated with distilled water than in those receiving disinfectants. Overall hatch of fertile eggs and egg moisture loss were comparable among all three treatments. At 1 day of age, the chicks incubated with 904 had a statistically lower yolk sac contamination rate than those incubated with 904+EDTA-Tris or distilled water. The 2-wk mortality rates, body weights, feed conversion ratios, yolk sac weights, and yolk sac contamination rates were all similar among the three treatments.

Colonization of the intestine of turkey embryos exposed to Mycoplasma iowae.

Eight-day-old turkey embryos were inoculated into the yolk sac with 3 X 10(5) colony-forming units of Mycoplasma iowae strain D112 in order to study the growth-depressing effect, the histopathological changes, and colonization of the intestinal tract. The embryo: egg weight ratio was significantly lower in the inoculated eggs than in controls. Histologically, there were infiltrations in parenchymatous organs and chorioallantoic membranes with heterophilic granulocytes. M. Iowae was demonstrated on the intestinal mucosa by antibody fluorescent microscopy and by transmission electron microscopy. Attaching mycoplasmas had a distinct morphology; the segment in contact with enterocytes was cone-shaped and had finely granulated cytoplasma which was abruptly separated from the distal coarsely granulated area. We conclude that M. iowae has a predilection for the intestinal tract of avian hosts.