Hatchability rate and embryonic growth of broiler chicks following in ovo injection royal jelly.

1. The objectives of this study were to compare the hatchability, chick body and internal organs weights and plasma testosterone concentration of hatchlings after in ovo administration of royal jelly (RJ) on d 7 of incubation. 2. Fertile eggs (n = 150) were injected into the air sac or yolk sac with 0.5 ml normal saline solution or normal saline and pure RJ. The eggs were randomly divided into 5 groups of 30 eggs each: NC, control eggs receiving no injection; ASA, air sac-injected eggs given normal saline solution; ARJ, air sac-injected eggs injected with pure RJ; YSA, yolk sac-injected eggs receiving normal saline solution and YRJ, yolk sac-injected eggs given pure RJ. 3. Injection of RJ significantly decreased hatchability (46.7%) compared with injection of saline solution (68.3%). Hatchability was lower in ARJ (33.3 %) and YRJ (60.0%) groups than in the NC group (90.0%). Hatchability in ASA (70.0%) and YSA (66.66%) groups were comparable to the NC group. 4. In ovo injection of RJ into both sacs increased chicks' absolute and relative body, heart, liver and testes weights compared to the control group whereas plasma testosterone concentration was similar among the different groups. 5. It was concluded that in ovo injection of RJ may be an effective method to increase the body weight of hatched chicks as an absolute value (CWT) and chicks' internal organ weights.

[The Ludwigshafen Osteoporosis Screening Questionnaire (LOS Questionnaire): result of the evaluation of anamnestic risk factors in osteoporosis diagnostics]. / Ludwigshafener Osteoporosescreeningbogen (LOS-Bogen) : Resultat aus der Evaluation anamnestischer Risikofaktoren in der Osteoporosediagnostik.

BACKGROUND: Osteoporosis is a major health problem worldwide and is included in the WHO list of the top ten major diseases. However, it is often undiagnosed until the first fracture occurs, due to inadequate patient education and lack of insurance coverage for screening tests. METHODS AND MATERIAL: In our study of 78 patients with metaphyseal long bone fractures, we searched for a correlation between anamnestic risk factors, bone-specific laboratory values, and the bone morphogenic density (BMD). Each indicator was examined as a possible diagnostic instrument for osteoporosis. The secondary aim of this study was to demonstrate the high prevalence of osteoporosis in patients with metaphyseal fractures. RESULTS: Of our fracture patients 76.9% had decreased bone density and 43.6% showed manifest osteoporosis in DXA (densitometry) measurements. Our modified LOS Questionnaire, identifying anamnestic risk factors, correlated highly significantly (p=0.01) with reduced BMD, whereas seven bone-specific laboratory values (p=0.046) correlated significantly. CONCLUSION: Anamnestic risk factors correlate with pathological BMD more than bone-specific laboratory values. The LOS Questionnaire used in this study would therefore function as a cost-effective primary diagnostic instrument for identification of osteoporosis patients.

Instant stem cell therapy: characterization and concentration of human mesenchymal stem cells in vitro.

In regenerative medicine, there is an approach to avoid expansion of the mesenchymal stem cell (MSC) before implantation. The aim of this study was to compare methods for instant MSC therapy by use of a portable, automatic and closed system centrifuge that allows for the concentration of MSCs. The main outcome measures were the amount of MSCs per millilitre of bone marrow (BM), clusters of differentiation (CD), proliferation and differentiation capacities of the MSC. A volume reduction protocol was compared to the traditional laboratory methods of isolation using a Ficoll gradient and native BM. Fifty millilitres of BM were obtained from haematologically healthy male Caucasians (n=10, age 8 to 49 years). The number of colony forming units-fibroblast (CFU-F)/ml BM was highest in the centrifuge volume reduction protocol, followed by the native BM (not significant), the centrifuge Ficoll (p=0.042) and the manual Ficoll procedure (p=0.001). The MSC of all groups could differentiate into the mesenchymal lineages without significant differences between the groups. The CD pattern was identical for all groups: CD13+; CD 44+; CD73 +; CD90+; CD105+; HLA-A,B,C+; CD14-; CD34-; CD45-; CD271-; HLA-DR-. In a further clinical pilot study (n=5) with 297 ml BM (SD 18.6), the volume reduction protocol concentrated the MSC by a factor of 14: there were 1.08 x 10(2) MSC/ml BM (standard deviation (SD) 1.02 x 10(2)) before concentration, 14.8 x 10(2) MSC/ ml BM (SD 12.4 x 10(2)) after concentration, and on average 296 x 10(2) MSC (SD 248.9 x 10(2), range 86.4-691.5 x 10(2)) were available for MSC therapy. The volume reduction protocol of the closed centrifuge allows for the highest concentration of the MSC, and therefore, is a promising candidate for instant stem cell therapy.

Effects of breed and progestin source on estrus synchronization and rates of fertility and fecundity in Iranian Sanjabi and Lori ewes.

A trial was conducted to evaluate the effects of FGA (Fluorogestone acetate) and CIDR (Controlled internal drug release) on the induction of estrus and pregnancy and fecundity rates of the Sanjabi and Lori sheep. A total of 360 Sanjabi and Lori sheep were randomly grouped into two treatments with intravaginal devices inserted for 13 days: Group FGA (40 mg FGA, n = 180) and Group CIDR (n = 180). All ewes received an i.m. injection of 400 IU eCG (equine chorionic gonadotrophin) at devices removal. Estrous was assessed by exposing all ewes to vasectomized rams at 12 h intervals. Cervical artificial insemination was performed 12 h after estrus onset. The overall estrus response was 72.5%. The source of progestin did not influence the efficiency of estrus response but a significant difference (p<0.05) was found between the breed groups (Lori: 88.6%, Sanjabi: 58.3%). Among the sheep that received either CIDR or FGA, estrus response was significantly (p<0.05) higher in the Lori (CIDR: 82.2%, FGA: 91.1%) than in the Sanjabi (CIDR: 64.4%, FGA: 52.2%) breed. The lambing and fecundity rates for all groups were 60.2% and 1.2 +/- 0.03, respectively. No significant differences in term of the lambing and fecundity rates were recorded between CIDR and FGA groups and among Lori and Sanjabi breed. The results of this study indicate the source of progestin or sheep breed did not influence the pregnancy and fecundity rates. The sheep breed influences the estrous response rate while the source of progestin did not affect the estrous response.

Symptomatic and asymptomatic cryptosporidiosis in young children in Iran.

The present study was conducted during the period May 2003 to October 2003 in 616 children less than three years of age with and without diarrheal disease presenting at the pediatric clinic of teaching hospitals in Kermanshah, Iran. Single stool specimens were collected from 515 diarrheic and 99 non-diarrheic children. Two smears were made from each stool samples and were stained by a modified Ziehl Neelsen technique. Cryptosporidium parvum (C. parvum) were detected in 10.4% of children. The C. parvum infection rate was significantly higher in diarrheic children (11.6%) than in non-diarrheic children (4.0%). C. parvum was observed more frequently in stool samples of children who lived in rural areas (15.2%) than those who lived in urban areas (7.2%). In regard to the presence of animals, the infection rate was 18.5% among children who lived in association with animals in comparison with 8.2% among those who lived in compounds with no animals. The majority of C. parvum cases occurred in children between the ages of 0-12 months (11.9%), followed by in children between the ages of 13-24 months (9.2%) and in children between the ages of 25-36 months (3.0%). The data suggest that C. parvum is relatively endemic in young children and that Cryptosporidium may be an important pathogen associated with diarrhea in young children.

Adherence of non-enteropathogenic Escherichia coli to HeLa cells.

Three hundred and nine strains of Escherichia coli isolated from infants and children with diarrhoea but not belonging to any recognised classes of diarrhoeagenic E. coli were investigated for their ability to adhere to HeLa cells in the presence of D-mannose. An enteroadherent-aggregative pattern (EAgg) was observed in 32.03%, localised adherence (LA) in 4.5%, diffuse adherence (DA) in 5.8%, and LA/DA and EAgg/LA in 1.9% and 1.2% of the isolates respectively. The results obtained with 100 control isolates were: EAgg 17%, LA 2%, DA 2%, LA/DA 2%, EAgg/LA 6% and DA/EAgg 1%. No adherence was manifested by 168 (54.36%) of 309 diarrhoeal isolates and 70% of the 100 control isolates. The results of this study showed that amongst non-enteropathogenic E. coli, strains exhibiting the EAgg pattern are significantly associated with diarrhoea (p < 0.005). Most of these strains showed a pattern of multiple drug resistance.

In vitro adhesion and invasion of Salmonella enterica serovar Havana.

A total of 132 isolates of the Havana serovar of Salmonela enterica were studied for their adhesion to and invasion of Hela cells in the presence of D-mannose. A total of 124 (93.9%) isolates were adherent, and 51% of these were invasive. Of these 124 strains 82 (66%) adhered to HeLa cells with a pattern typical for enteroaggregative (EAgg) Escherichia coli whereas the remaining 34% adhered in a non-aggregative (non-EAgg) pattern. Of the 82 strains showing the EAgg pattern 60% were invasive whereas only 33% of the 49 non-EAgg isolates invaded HeLa cells (P < 0.01). The action of mannose-sensitive type 1 pili was inhibited by the addition of D-mannose. Therefore we conclude that type 1 pili are not important in adhesion and invasion of Havana.

The use of biochemical fingerprinting, phage typing and antimicrobial-susceptibility testing in the detection of epidemic strains of Salmonella of serotype Typhimurium in Iran.

A collection of 86 strains of Salmonella of serotype Typhimurium isolated from children with gastroenteritis in Tehran, Iran was examined for biochemical phenotype, phage type and antibiotic-resistance pattern. Twenty-seven biochemical phenotypes (BPTs), 14 discrete phage types (PTs) and 18 resistotypes (RTs) were identified. Fifty-three strains (62%) belonged to two major and probably related BPTs, whereas the other 33 isolates belonged to less common BPTs. The two predominant BPTs contained 26 strains of the same PT and 23 strains of the same RT. Different PTs and RTs of strains with similar BPT were sometimes observed, possibly reflecting antibiotic pressures in Iran. These results suggest that two major "clones" of Typhimurium strains are particularly common in Iran and, although each method alone adequately detected these and other less common "clones", biochemical fingerprinting provided additional information about relationships among strains.

Application of biochemical fingerprinting to the investigation of clonal groups of Salmonella of serotype Havana.

A computerised typing method based on biochemical fingerprinting was used to investigate biochemical phenotypes (BPTs) among 70 strains of Salmonella of serotype Havana isolated from human cases of gastroenteritis in Iran and other parts of the world. A total of 16 BPTs comprising five common and 11 single phenotypes was identified. The most frequently found BPT contained 24 isolates from Iran and nine from other countries. Three common BPTs with two, seven and 15 isolates were found among Iranian strains only and one common BPT with two isolates was found among non-Iranian strains only. Antibiotic-resistance patterns and virulence properties of strains from these common BPTs suggested that they might be unique clones. Forty-two Iranian isolates shared multi-resistance to between three and seven antibiotics. In contrast, none of the isolates from other countries was resistant to antibiotics. Furthermore, 43 Iranian isolates showed mannose-resistant adhesion to HeLa cells and 24 of them possessed an aerobactin-mediated iron-uptake system, whereas none of the isolates from other countries possessed any of these virulence properties. These findings suggest that four unique clones of Salmonella Havana with different BPTs and virulence properties are common in Iran; two particular clones were responsible for a majority of Havana infections there. However, the most prevalent BPT found among Iranian strains was also common in strains from other countries. It is concluded that biochemical fingerprinting, as used in this study, is a reliable method for identifying clonal groups of Havana strains. The method is reproducible, easy to perform and can be used alone, or in combination with other typing methods, in epidemiological studies of serotype Havana.

Occurrence of colonization factor antigens I & II in enterotoxigenic Escherichia coli associated diarrhoea in Iran & correlation with severity of disease.

The occurrence of colonization factor antigens I and II (CFA/I and II) and type 1 somatic pili was investigated in 197 enterotoxigenic Esch. coli (ETEC) isolated from 197 patients of diarrhoea (aged under 3 yr) during February 1985 to March 1986 in Tehran, Iran. Among ETEC strains, 154 strains were heat-stable enterotoxin (ST) producers, 27 strains were heat-labile enterotoxin (LT) producers, and 16 strains produced both toxins. Sixty five (33%) strains showed mannose-resistant haemagglutination (MRHA) of human and/or bovine erythrocytes; of these, 51 (86%) strains were positive for CFA/I and II. Seventy one (36%) strains also exhibited type 1 somatic pili. CFA/I was found in 4 (15%) LT producing, 24 (16%) ST producing, and 2 (13%) LT/ST producing strains. In contrast, CFA/II was only found in ST producing strains (17 strains) and those producing both toxins (4 strains). Patients having CFAs-positive ETEC strains had a significantly (P less than 0.001) higher number of stool evacuation per day and a longer duration of diarrhoea than those having CFAs-negative strains. Fifty nine patients had mixed infections of ETEC strains and other enteropathogens. CFA/I or II (CFAs)-positive and CFAs-negative ETEC strains were found in 17 and 42 patients with mixed infections respectively. The mean number of stool evacuations per day was much higher in patients with ETEC and rotavirus than those with only ETEC infection (P less than 0.001). However, severity of the disease was not affected by the presence or absence of CFA/I or II in ETEC strains found in these patients.

Aetiological studies of diarrhoeal diseases in infants and young children in Iran.

The aetiology of diarrhoeal diseases was investigated among 715 patients admitted to four Children's Hospitals in Tehran, during February 1986 to March 1987, and also among 443 patients attending the central Out-Patient Clinic in Sanandaj, State of Kordestan, during October 1986. Enteropathogenic Escherichia coli (EPEC) were the most common pathogens found in both areas. Almost 26.7% of the patients in Tehran and 20.1% of the patients in Sanandaj were infected with EPEC. Enterotoxigenic E. coli (ETEC) were the next most frequent groups found (17.1% both in Tehran and Sanandaj), with heat-stable enterotoxin (ST)-producing strains being dominant. Of 122 ETEC strains isolated in Tehran, 94 (77%) strains produced ST, 15 (12.3%) strains produced heat-labile enterotoxin (LT) and 13 (10.7%) strains produced both LT and ST. Almost the same pattern of toxigenicity was observed among ETEC strains isolated in Sanadaj. Of the 76 ETEC strains isolated in this area, 70 (92.1%) strains were ST producers, followed by those producing both LT and ST (five strains) and LT only (one strain). One strain of enteroinvasive E. coli (EIEC) was also isolated from a patient in Tehran. The rates of salmonella and shigella isolation were 8.8 and 5.7% in Tehran and 3.8 and 4% in Sanandaj respectively. Enterohaemorrhagic E. coli, Vibrio cholerae and V. parahemolyticus were not isolated but a mixture of two or more pathogens was found in 59 patients (8.2%) in Tehran and in 20 patients (4.5%) in Sanandaj. These findings suggest that diarrhoegenic E. coli are the most important cause of diarrhoeal diseases in infants and young children in these areas in Iran.

Antimicrobial drug resistance and resistance factor transfer among clinical isolates of salmonellae in Iran.

508 salmonella strains isolated during 1983-1986 in Tehran, Iran from cases of diarrhoea in children less than 5 were tested for sensitivity to 10 antimicrobial drugs and their ability to transfer the resistance determinants. Salmonella typhimurium and S. havana were most common (45.7% and 30.9%, respectively) followed by S. typhi (2.9%) and S. larochelle (2.4%). The antimicrobial agents used were chloramphenicol (C), gentamicin (G), tetracycline (T), ampicillin (A), cephalothin (Cf), trimethoprim-sulfamethoxazole (SXT), kanamycin (K), nalidixic acid (Na), streptomycin (S), and furoxone (F). 91% of the salmonellae isolates contained resistance determinants, of which 89.6% were resistant to more than one agent. Strains resistant to 4-7 drugs comprised 85.6% of the total isolates. Ampicillin resistance was found in 85.4% and nalidixic acid in 2.7%. 58 different patterns of resistance to the antimicrobial agents used were observed, of which C/T/A/SXT/K/S and C/T/A/SXT/K/S/Cf were the most frequently encountered. 71.9% of the resistant strains contained transferable resistance factors and S. typhimurium had the highest rate of transfer (80.8%). Resistance to chloramphenicol had the highest rate of transfer among the agents used (77.4%) and streptomycin the lowest (20.0%). The pattern most frequently transferred was C/T/A/SXT/K (41.9%).