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Detection of the mgtC gene in multidrug-resistant Salmonella sp. based on isolation of chicken eggshell swabs from traditional Surabaya markets.

BACKGROUND: The virulence of Salmonella sp. is increased by the presence of the mgtC gene, which allows the bacteria to survive in environments with low magnesium levels, such as inside macrophages. Salmonella sp. found on eggshells when they show resistance to three or more classes of antibiotics can be classified as multidrug-resistant (MDR) bacteria. AIM: This study aimed to identify the presence of Salmonella sp. MDR and the mgtC gene in chicken eggshell swabs from traditional markets in Surabaya. METHODS: Swab samples were collected from 160 eggs (80 from layer chickens and 80 from free-range chickens) at 10 traditional markets in Surabaya, Indonesia. Isolation and identification were performed using culture media, including Salmonella Shigella Agar, Gram staining, Triple Sugar Iron Agar, Sulfide Indole Motility, Simmons Citrate Agar, Methyl Red and Voges Proskauer, and Urea Agar. Antibiotic sensitivity testing was performed using the disc diffusion method on Mueller-Hinton Agar. Molecular detection of the mgtC gene was performed using polymerase chain reaction. RESULTS: The results showed that 16.87% (27/160) of the samples were detected positive for Salmonella sp. All Salmonella sp. isolates (27) were resistant to Erythromycin (100%). Resistance was also found to Ampicillin (77.77%, 21/27), Tetracycline (29.62%, 8/27), and Ciprofloxacin (18.51%, 5/27). No resistance to chloramphenicol was observed. In addition, eight of the 27 isolates (29.62%) were classified as Salmonella sp. MDR. The Salmonella sp. MDR isolates also carried the mgtC gene at 87.5% (7/8). CONCLUSION: These findings demonstrate the potential global public health threat posed by MDR Salmonella sp. with the mgtC gene, emphasizing the importance of monitoring and controlling antibiotic resistance in humans and animals.

Animals↗

Efficacy of a live avirulent Salmonella typhimurium vaccine in preventing colonization and invasion of laying hens by Salmonella typhimurium and Salmonella enteritidis.

An avirulent live delta cya delta crp Salmonella typhimurium strain chi 3985 that precludes colonization and invasion of chickens by homologous and heterologous Salmonella serotypes was evaluated for its long-term protection efficacy. Chickens vaccinated orally at 2 and 4 wk of age were assessed for protection against oral challenge with wild-type S. typhimurium and Salmonella enteritidis strains at 3, 6, 9, and 12 mo of age. A comparison of Salmonella isolation from vaccinated and nonvaccinated layers after challenge with S. typhimurium or S. enteritidis showed that delta cya delta crp S. typhimurium chi 3985 induced excellent protection against intestinal, visceral, reproductive tract, and egg colonization, invasion, and/or contamination by Salmonella. The duration of protection lasted for 11 mo after vaccination, at which time the experiment was terminated. S. enteritidis and S. typhimurium were isolated from the yolk, albumen, and shells of eggs laid by nonvaccinated chickens challenged with Salmonella. S. typhimurium caused pathological lesions in nonvaccinated chickens, whereas vaccinated and nonvaccinated chickens challenged with S. enteritidis showed no pathological lesion in the visceral and reproductive organs. Vaccination with chi 3985 prevented transmission of S. typhimurium or S. enteritidis into eggs laid by vaccinated layers with no effect on egg production. To our knowledge, this is the first publication confirming that vaccination with live avirulent Salmonella can induce long-term protection against Salmonella infection in layers.

Administration, Oral↗

Effect of DDE on the Ca metabolism of the duck eggshell gland and its subcellular fractions; relation to the functional stage.

Administration of DDE to Indian Runner ducks (Anas platyrhynchos var.) reduced the eggshell index by 28% and the calcium content of the secreted fluid in the shell gland by 25%. DDE inhibited the functional decrease of the calcium content of the gland cells that occurred during eggshell formation. DDE reduced the functional increase in the ATP-dependent binding of Ca2+ to a microsomal subfraction FIII prepared from Ca-secreting glands forming an eggshell. In subfraction FI the Ca2+ binding was decreased by DDE even when the gland contained a mature shell. FIII probably contained calcium-secreting granules, whereas FI contained plasma membrane fragments of the gland cells.

Animals↗

Adult Coturnix quail bronchitis.

Quail bronchitis was confirmed by recovery of fowl adenovirus serotype 1 from 2 flocks of adult birds (Coturnix coturnix japonica) that were reared commercially for their eggs in the Republic of Singapore. Both flocks were reported to have a fall in egg numbers of 10-15%. The main clinical signs were respiratory distress and soft-shelled and white eggs lacking shell pigmentation.

Adenoviridae Infections↗

Transmission electron microscopy of the vertical crystal layer and cuticle of the eggshell of the domestic fowl.

1. Eggshell pieces (1 cm2) were decalcified using a solution of EDTA (200 g/litre, pH 6.9 to 7.0) in paraformaldehyde (2 g/litre) and glutaraldehyde (0.5 g/litre) in phosphate buffer. 2. They were prepared for transmission electron microscopy (TEM). 3. TEM identified a vertically aligned matrix associated with the vertical crystal layer (VCL). It is hypothesised that the vertical orientation of calcite crystals in the VCL is closely linked to this vertical matrix. 4. TEM also revealed the presence of a 2-layered cuticle, the inner layer containing vesicles which were absent in the outer. 5. Cuticular vesicles contain hydroxyapatite and are thought to play a role in the termination of shell formation. The current paper presents data relating to microbial apatitic systems that strengthen this hypothesis.

Animals↗

Parathyroid hormone receptor binding property in the shell gland of oviduct of the guineafowl during an oviposition cycle.

Parathyroid hormone (PTH) receptor binding property in the membrane fraction of the endometrium of the shell gland (uterus) of the guineafowl was analyzed by the use of [125I]PTH-related peptide (PTHrP) binding assays. Specificity, reversibility, and saturation of binding were demonstrated. Scatchard plots revealed a single class of binding sites. The equilibrium dissociation constant (Kd) was 0.50 to 1.15 nM in laying birds and 1.07 to 1.16 nM in nonlaying birds. The maximum binding capacity (Bmax) per milligram of membrane protein was 65.2 to 110.9 femtomoles (fmol) in laying birds and 105.8 to 120.6 fmol in nonlaying birds. Both Kd and Bmax values changed within the above range during an oviposition cycle in laying birds, showing a decrease during the period of eggshell formation. No change was found in nonlaying birds. The results suggest that PTH/PTHrP receptors are present in the shell gland of the guineafowl, and their binding may be related to eggshell formation.

Animals↗

The modifications of cortical endoplasmic reticulum during in vitro maturation of Xenopus laevis oocytes and its involvement in cortical granule exocytosis.

In Xenopus laevis eggs, cisternae shells which surround cortical granules (CG) are part of a cortical endoplasmic reticulum (ER) network. In this paper the origin of such ER shells has been studied in full-grown, progesterone-exposed Xenopus oocytes. Furthermore, the possible role of the cortical ER in the activation process has been investigated by pricking maturing oocytes. It has been shown that in full-grown ovarian oocytes ER CG shells are absent and ER cisternae are extensively and randomly distributed throughout the peripheral cytoplasm, where they appear to be continuous with annulate lamellae (AL). Following hormone treatment, the AL completely disaggregate and the ER cisternae gradually migrate to the cortex where they surround the CG constituting the typical cortical network described in uterine eggs. Furthermore, it has been found that 8 h after progesterone treatment (before the first polar body extrusion) the response to pricking (CG exocytosis) occurs only at the animal half; there is no observable response in the vegetal half. At this time ER shells surround CG only in the animal hemisphere. A complete CG exocytosis occurs following the first polar body emission, when the cortical ER is well organized in the whole oocyte cortex. The correlation between the differentiation of the cortical ER and the arousal in the oocyte of the ability to respond to a pricking stimulus is discussed in the light of an involvement of the cortical ER in the propagation of CG exocytosis.

Animals↗

Relationship between shell structure and movement of Salmonella enteritidis across the eggshell wall.

1. Ultrastructural analyses of the eggshells of one strain of commercial layers indicated that the cuticular layer is rarely present as an even covering over the shell surface and the paired shell membranes are invariably pitted with holes larger than bacterial dimensions. 2. The above conditions pertain irrespective of the age of the laying bird. 3. Bacterial penetration of the eggshell is independent of pore numbers. 4. In the absence of a functional cuticle and with the shell membranes removed, bacteria are checked in their movement by structural modifications in the mammillary layer.

Age Factors↗

Organic matrix morphology and distribution in the palisade layer of eggshells sampled at selected periods during lay.

1. 1 cm2 pieces of eggshells from a commercial battery flock were plasma etched to remove the outer shell membranes. 2. They were decalcified using EDTA (200 g/l, pH 6.9 to 7.0) in paraformaldehyde (20 g/l) and 25% gluteraldehyde (20 ml in 0.98 l) in phosphate buffer, then prepared for light and transmission electron microscopy. 3. Light microscopy revealed a differential distribution of matrix material within all 3 regions of the palisade layer at the beginning of lay. 4. Transmission electron microscopy revealed a more even distribution of matrix at the beginning of lay, although morphological differences were observed. At the end of lay all 3 regions showed an increase in % matrix and vesicles/10 cm2 of micrograph compared to the middle and beginning of lay periods. 5. It is hypothesised that matrix vesicles are involved in the regulation of the physiochemical environment within the forming eggshell and that the decline in shell quality associated with the end of lay is related to a concomitant change in matrix quality.

Animals↗

Expression of the parathyroid hormone-related protein gene in the avian oviduct: potential role as a local modulator of vascular smooth muscle tension and shell gland motility during the egg-laying cycle.

The phylogenetic conservation of the primary structure of PTH-related protein (PTHrP) supports an important, yet undetermined, role(s) for this molecule in the biology of birds and mammals. As an initial step toward understanding the function of PTHrP in birds, we investigated the expression of PTHrP mRNA in tissues of the egg-laying hen. This analysis revealed that PTHrP mRNA is expressed at various levels in lung, brain, heart, and tissues of the digestive tract, including the proventriculus (secretory stomach), gizzard, and small intestine. In the oviduct tissues of adult birds, PTHrP mRNA was detected in the isthmus (membrane-secreting) and shell gland (calcium-secreting) portions, but not in magnum (albumin secreting) tissue. During oviduct development, high levels of PTHrP mRNA present in the oviducts of the 12-week-old bird suggest a role for PTHrP in oviduct development. Interestingly, as the oviduct matures, relatively high levels of PTHrP mRNA segregate with the distal tissues that ultimately differentiate into the isthmus and shell gland (uterus). To address a possible role for PTHrP in the differentiated function of the shell gland, we followed the expression of PTHrP in the shell gland at different times in the laying cycle and found levels of PTHrP to transiently increase as the egg moves through the oviduct, gradually returning to basal levels in the 15-h calcification period. We localized the cycle-associated fluctuations in PTHrP mRNA levels to the shell gland serosa and smooth muscle layer. Immunoreactive PTHrP was localized to the serosal membrane as well as the smooth muscle layer of serosal arterioles, suggesting that PTHrP may modulate vascular smooth muscle activity. In support of this hypothesis, synthetic chicken PTHrP (1-34)NH2 was found to relax the resting tension of isolated shell gland blood vessels in a dose-dependent manner. Together, these data indicate that the expression of the PTHrP gene in the avian oviduct is both temporally and spatially regulated during the egg-laying cycle and that PTHrP may function as an autocrine/paracrine modulator of shell gland smooth muscle activity of both ductal and vascular origins. The vasorelaxant property of N-terminal fragments of PTHrP supports a role for this molecule in the temporal increase in blood flow to the shell gland during egg calcification.

Animals↗

Progesterone stimulates prostaglandin synthesis in eggshell gland mucosa of estrogen-primed chickens.

1. Prostaglandins may be involved in calcium translocation in the avian shell gland, since indomethacin, administered at the beginning of shell formation, reduces eggshell thickness as well as 45Ca-uptake and prostaglandin synthesis by a homogenate of eggshell gland mucosa. 2. The stimulus for calcium transport in the shell gland during shell formation remains unknown. 3. The present study was undertaken to investigate the effects of progesterone on prostaglandin formation by the eggshell gland mucosa of the domestic fowl. 4. Progesterone significantly stimulated synthesis of PGF2 alpha, PGE2 and TXB2 by eggshell gland mucosa homogenate. 5. Progesterone treatment also induced the synthesis of the biotin-binding protein, avidin. 6. A microsomal fraction prepared from the eggshell gland mucosa had a high affinity for binding PGE2. 7. Progesterone treatment reduced the KD value of this binding without affecting the maximal number of binding sites. 8. Progesterone did not change the total calcium content of shell gland mucosa. 9. The role progesterone plays in prostaglandin formation and calcium transport in the eggshell gland mucosa is discussed.

Animals↗

Determination of 4,4'-dinitrocarbanilide (DNC), a component of Nicarbazin, in Canada goose (Branta canadensis) eggshells using high-performance liquid chromatography.

A method was developed using high-performance liquid chromatography to assay 4,4'-dinitrocarbanilide (DNC), the active ingredient in Nicarbazin, in eggshells collected from Canada geese fed a formulated feed fortified with Nicarbazin at doses of 0, 125, 250, and 500 microg/g. The method was developed using chicken eggshells fortified with DNC. The method was used to quantify DNC in both the shell-associated membranes and the calcified shell extracellular matrix. These values were compared to those obtained for a composite sample consisting of both the membranes and the calcified shell extracellular matrix. The validated method was used to quantify DNC in eggshells from geese fed fortified feed to ascertain the effect of Nicarbazin feed concentration on shell DNC concentration. DNC levels in the eggshells were highly correlated with feed dose.

Animal Feed↗

Changes in egg production rate induced by progesterone injection in broiler breeder hens.

A polycystic ovarian follicle (PCOF) syndrome associated with high baseline concentrations of progesterone (P4) without preovulatory luteinizing hormone (LH) surges has been reported in turkey hens. The PCOF syndrome could be induced in turkey hens by injecting P4 (0.33 mg/kg per d) daily early in the reproductive period for 10 to 12 d and then waiting 3 wk for the syndrome to develop. It was hypothesized that an arrest in laying associated with the PCOF syndrome could be induced by daily injection of P4 in restricted-fed broiler breeder hens. Hens were divided into 5 treatment groups and were injected subcutaneously daily with P4 in canola oil at dosages of 0, 0.17, 0.33, 0.5, and 1.5 mg/kg per d for 13 d, at 14 wk of egg production when they were 41 wk of age. Blood samples were collected on d 7 and 13 immediately before P4 injection. Oviductal and ovarian morphologies were measured at necropsy 1 d after the last P4 injection. Egg production rate was reduced by injection of P4 at dosages < 0.17 mg/kg per d. At dosages of 0.5 and 1.5 mg/kg per d, ovarian hierarchical follicles had regressed. None of the broiler breeder hens had the PCOF syndrome at necropsy, but a high incidence of hens holding hard-shelled uterine eggs for several days was observed. Concentrations of LH decreased with P4 injection at > 0.17 mg/kg per d, and P4 concentrations were increased with P4 injection at > 0.5 mg/kg per d. Estradiol-17beta (E2) concentrations were decreased at all P4 dosages. It was concluded that egg production rate was reduced by daily injection of P4 at dosages > 0.17 mg/kg per d, and egg production ceased and ovarian follicles and the oviduct regressed at dosages > 0.50 mg/kg per d. The PCOF syndrome, however, was not induced in restricted-fed broiler breeder hens by P4 injection.

Animals↗

Supplemented eggshell restores calcium transport in chorioallantoic membrane of cultured shell-less chick embryos.

It was previously reported (Tuan, 1980a) that the development-specific expression of calcium transport and related functions in the chick embryonic chorioallantoic membrane (CAM) requires the continuous presence of the eggshell, the calcium source of the embryo. To further understand the mechanism of action of the eggshell on the CAM functions, this study reports the effects of eggshell supplementation on chick embryos maintained in shell-less cultures. The cultured embryos were able to accumulate and utilize the exogenous shell calcium, applied directly onto the CAM, for skeletal formation. In the region of the CAM directly adhering to the added shell, calcium transport activity, calcium-binding protein (CaBP) activity, and vitamin K-dependent gamma-glutamyl carboxylase activity were significantly restored. These results strongly suggest that the proximity of shell calcium may regulate expression of calcium transport and related functions in the chick embryonic CAM.

Allantois↗

Cholesterol concentration of egg yolk and blood plasma and performance of laying hens as influenced by dietary alpha-ketoisocaproic acid.

Experiments were conducted to determine the effect of supplemental alpha-ketoisocaproic acid (KIC) or Leu on layer performance and plasma and egg cholesterol levels. In the first experiment, 0, .09, or .27% KIC and .09 or .27% Leu were fed to 62-wk-old laying hens. In a second experiment, the same levels plus another level of .54% KIC and .54% Leu were fed to 32-wk-old hens for an 8-wk period. The basal diet or the basal diet supplemented with .27% KIC or .27% Leu were continued for 20 wk to determine the long-term effects of these supplements. In a third experiment, hens were fed 0, .1, .2, or .4% KIC for 8 wk. The addition of KIC and Leu to the hen diets did not affect percentage egg production, egg weight, shell thickness, or Haugh units. In Experiment 1, .27% KIC and .09% Leu significantly increased BW gain. Neither KIC or Leu significantly affected BW gain in Experiment 2 when measured at 8 or 28 wk. In the third experiment, diets supplemented with KIC increased significantly BW gain. In Experiment 1, .09% KIC and .09% Leu significantly reduced egg cholesterol at 4 wk. After 8 wk, .27% KIC reduced egg cholesterol significantly below the controls. No effect on plasma or egg cholesterol was observed in Experiments 2 and 3.

Animal Feed↗

Hymenolepis microstoma: direct life cycle in immunodeficient mice.

The mouse bile duct tapeworm Hymenolepis microstoma requires beetles as the obligatory intermediate host. However, when congenitally athymic NMRI-nu mice were infected with the mature tapeworm and allowed to eat their own faeces with tapeworm eggs, the oncospheres penetrated the intestinal tissue and developed to cysticercoids. After excysting, growth to adult worms occurs in the lumen of the small intestine and bile duct. Furthermore, the same happened when NMRI-nu mice, non-obese diabetic severe combined immunodeficiency (NOD/Shi-scid) mice and NOD/Shi-scid, IL-2 Rgamma(null) (NOG) mice were orally inoculated with shell-free eggs of this parasite. Differences between the cysticercoids of H. microstoma and H. nana developed in the mouse intestinal tissues were: (i) the time course for the development of fully matured cysticercoids of H. microstoma in mice was about 11 days but only 4 days for H. nana; and (ii) cysticercoids of H. microstoma developed in mice had a tail while those of H. nana had none.

Animals↗

Measurement of T-2 and HT-2 toxins in eggs by high-performance liquid chromatography with fluorescence detection.

T-2 toxin is a mycotoxin produced by several species of common fungi capable of infesting human food and animal feeds. Lower-quality feeds given to chickens may be contaminated with T-2 toxin, which may affect their health. The literature suggests that T-2 toxin is transmitted from the hen to the eggs. This article describes the development of a liquid chromatographic assay for T-2 and the related mycotoxin HT-2 in eggs. T-2 and HT-2 toxins were isolated from spiked eggs with a tandem charcoal-alumina-Florisil column and immunoaffinity column cleanup. The isolated toxins were derivatized with the fluorophore 1-anthroyl nitrile, separated by high-performance liquid chromatography, and quantitated by fluorescence. The limit of detection of the method was 1 ng ml(-1) (parts per billion) of T-2 and HT-2 in whole (with shell removed) eggs. The limit of quantitation for both toxins was 5 ng ml(-1). Recoveries from spiked eggs over the range from 5 to 50 ng ml(-1) averaged 89.2% for T-2 and 100.3% for HT-2, with coefficients of variation of 3.5 and 8.2%, respectively. This method is sensitive enough to be used to check for the presence of T-2 or HT-2 toxins in eggs.

Animals↗

Transfer of orally administrated iodine-131 into chicken eggs.

Radioactive iodine-131 as both as free iodide (Na131I) and covalently bound to aniline (aniline-131I) was added to the drinking water of two Leghorn laying hens as a single dose and also as a cumulative dose over 1 week. The radioactivity of the principal parts of the eggs, i.e. shell, white, and yolk, was measured, and the radioactivity levels per gram material, and percent of the total radioactivity were calculated. The radioactivity measurements were continued for 1 month following the administration of 131I. In the case of the single dose administration, the results obtained showed that about 15% of the total radioactivity administered as Na131I was transported into the egg structure; compared to only about 1% for aniline-131I. After cumulative administration, about 15% of the total administered radioactivity was transported into the egg structure with both forms of 131I. This was probably because of metabolic cleavage of iodine bonds in the labeled aniline molecules during the longer period of exposure. These results also showed considerable accumulation of 131I in the egg yolks. In the case of the single dose administration, 131I can be detected in eggs up to about 20 days after administration, and up to about 30 days, in the case of the cumulative administration over 1 week.

Administration, Oral↗