NRASQ61R mutation drives elevated angiopoietin-2 expression in human endothelial cells and a genetic mouse model.

BACKGROUND: Angiopoietin-2 (Ang-2) is increased in the blood of patients with kaposiform lymphangiomatosis (KLA) and kaposiform hemangioendothelioma (KHE). While the genetic causes of KHE are not clear, a somatic activating NRASQ61R mutation has been found in the lesions of KLA patients. PROCEDURE: Our study tested the hypothesis that the NRASQ61R mutation drives elevated Ang-2 expression in endothelial cells. Ang-2 was measured in human endothelial progenitor cells (EPC) expressing NRASQ61R and a genetic mouse model with endothelial targeted NRASQ61R. To determine the signaling pathways driving Ang-2, NRASQ61R EPC were treated with signaling pathway inhibitors. RESULTS: Ang-2 levels were increased in EPC expressing NRASQ61R compared to NRASWT by Western blot analysis of cell lysates and ELISA of the cell culture media. Ang-2 levels were elevated in the blood of NRASQ61R mutant mice. NRASQ61R mutant mice also had reduced platelet counts and splenomegaly with hypervascular lesions, like some KLA patients. mTOR inhibitor rapamycin attenuated Ang-2 expression by NRASQ61R EPC. However, MEK1/2 inhibitor trametinib was more effective blocking increases in Ang-2. CONCLUSIONS: Our studies show that the NRASQ61R mutation in endothelial cells induces Ang-2 expression in vitro and in vivo. In cultured human endothelial cells, NRASQ61R drives elevated Ang-2 through MAP kinase and mTOR-dependent signaling pathways.

A machine learning model to predict therapeutic inertia in type 2 diabetes using electronic health record data.

OBJECTIVE: To estimate the therapeutic inertia prevalence for patients with type 2 diabetes, develop and validate a machine learning model predicting therapeutic inertia, and determine the added predictive value of area-level social determinants of health (SDOH). METHODS: This prognostic study with a retrospective cohort design used OneFlorida data (linked electronic health records (EHRs) from 1240 practices/clinics in Florida). The study cohort included adults (aged ≥ 18) with type 2 diabetes, HbA1C ≥ 7% (53 mmol/mol), ≥one ambulatory visit, and ≥one antihyperglycemic medication prescribed (excluded patients prescribed insulin before HbA1C). The outcome was therapeutic inertia, defined as absence of treatment intensification within six months after HbA1C ≥ 7% (53 mmol/mol). The predictors were patient, provider, and healthcare system factors. Machine learning methods included gradient boosting machines (GBM), random forests (RF), elastic net (EN), and least absolute shrinkage and selection operator (LASSO). The DeLong test compared the discriminative ability (represented by C-statistics) between models. RESULTS: The cohort included 31,087 patients with type 2 diabetes (mean age = 58.89 (SD = 13.27) years, 50.50% male, 58.89% White). The therapeutic inertia prevalence was 39.80% among the 68,445 records. GBM outperformed (C-statistic from testing sample = 0.84, 95% CI = 0.83-0.84) RF (C-statistic = 0.80, 95% CI = 0.79-0.80), EN (C-statistic = 0.80, 95% CI = 0.80-0.81), and LASSO (C-statistic = 0.80, 95% CI = 0.80-0.81), p < 0.05. Area-level SDOH significantly increased the discriminative ability versus models without SDOH (C-statistic for GBM = 0.84, 95% CI = 0.84-0.85 vs. 0.84, 95% CI = 0.83-0.84), p < 0.05. CONCLUSIONS: Using EHRs of patients with type 2 diabetes from a large state, machine learning predicted therapeutic inertia (prevalence = 40%). The model's ability to predict patients at high risk of therapeutic inertia is clinically applicable to diabetes care.

Kaposiform lymphangiomatosis: Diagnosis, pathogenesis, and treatment.

Kaposiform lymphangiomatosis (KLA) is a life-threatening rare disease that can cause substantial morbidity, mortality, and social burdens for patients and their families. Diagnosis often occurs long after initial symptoms, and there are few centers in the world with the expertise to diagnose and care for patients with the disease. KLA is a lymphatic anomaly and significant advancements have been made in understanding its pathogenesis and etiology since its first description in 2014. This review provides multidisciplinary, comprehensive, and state-of-the-art information on KLA patient presentation, diagnostic imaging, pathology, organ involvement, genetics, and pathogenesis. Finally, we describe current therapeutic approaches, important areas for research, and challenges faced by patients and their families. Further insights into the pathogenesis of KLA may advance our understanding of other vascular anomalies given that similar signaling pathways may be involved.

Amiodarone Extraction by the Extracorporeal Membrane Oxygenation Circuit.

Amiodarone is an anti-arrhythmic agent that is frequently used to treat tachycardias in critically ill adults and children. Because of physicochemical properties of amiodarone, extracorporeal membrane oxygenation (ECMO) circuits are expected to extract amiodarone from circulation, increasing the risk of therapeutic failure. The present study seeks to determine amiodarone extraction by the ECMO circuit. Amiodarone was administered to three ex vivo circuit configurations (n = 3 per configuration) to determine the effect of each circuit component on drug extraction. The circuits were primed with human blood; standard amiodarone doses were administered; and serial samples were collected over 24 hours. Additional circuits were primed with crystalloid fluid to analyze the effect of blood on extraction and to investigate circuit saturation by drug. The crystalloid circuits were dosed multiple times over 72 hours, including a massive dose at 48 hours. For both setups, the flow was set to 1 L/min. Drug was added to separate tubes containing the prime solution to serve as controls. Drug concentrations were quantified with a validated assay, and drug recovery was calculated for each sample. Mean recovery for the circuits and controls were compared to correct for drug degradation over time. Amiodarone was heavily extracted by all ECMO circuit configurations. Eight hours after dosing, mean recovery in the blood prime circuits was 13.5-22.1%. In the crystalloid prime circuits, drug recovery decreased even more rapidly, with a mean recovery of 22.0% at 30 minutes. Similarly, drug recovery decreased more quickly in the crystalloid prime controls than in the blood prime controls. Saturation was not achieved in the crystalloid prime circuits, as final amiodarone concentrations were at the lower limit of quantification. The results suggest that amiodarone is rapidly extracted by the ECMO circuit and that saturation is not achieved by standard doses. In vivo circuit extraction may cause decreased drug exposure.

Roles of the sire and dam quail in egg, yolk, albumen, and shell weight alterations due to the parthenogenetic trait.

Infertile eggs from Chinese Painted quail exhibit parthenogenesis, and the 1st egg in a clutch sequence is more likely to develop a parthenogenetic embryo. Also, hens that exhibit parthenogenesis have shorter clutches and heavier egg weights. These larger eggs may be a result of the egg residing in the oviduct longer, allowing more time for the parthenote to develop. However, it is unknown which components of the egg are altered due to the parthenogenetic trait or the role of dams and sires from the parthenogenetic line of quails in these alterations. Therefore, our objective was to determine if the parthenogenetic trait in dams, sires, or both contributes to alterations in egg components, such as yolk, albumen, and shell weights. Two selected lines of quail, 1 line selected for parthenogenesis and 1 line that was unselected for the parthenogenetic trait (control), were utilized in a 2 x 2 factorial arrangement of dam and sire lines to create 4 breeding treatments: control dams + control sires (CC), control dams + parthenogenetic sires (CP), parthenogenetic dams + control sires (PC), parthenogenetic dams + parthenogenetic sires (PP). Daily, fresh eggs were collected, labeled, weighed, and the germinal disc was classified as fertile, unfertilized without development, or parthenote. Yolk, albumen, and shell weights were obtained, and their weights were also calculated as percentage of total egg weight. A dam main effect revealed heavier weights for total egg, yolk, albumen, and shell as well as a larger percentage of albumen and ratio of albumen to yolk in parthenogenetic line dams versus control line dams. However, the percentage of yolk was higher for control than parthenogenetic line dams. The increase in total egg and egg component weights due to the parthenogenetic trait suggests that the transit time of the egg through the oviduct is altered. Perhaps transit through the magnum and uterus is delayed the most yielding greater albumen and shell weights, respectively.

Parental sex effect of parthenogenesis on progeny production and performance of Chinese Painted Quail (Coturnix chinensis).

Embryonic development of an unfertilized egg, parthenogenesis, is known to occur in Chinese Painted quail. However, selection for parthenogenesis in both the dam and sire leads to a reduction in hatchability following mating. Therefore, the objective of this study was to determine if selection for parthenogenesis in the dam, sire, or both also impact their progeny performance. There were 2 lines of birds used in this trial: 1 line selected for parthenogenesis and 1 line not selected for parthenogenesis (control) yielding breeding pairs as follows: control dams + control sires (CC), control dams + parthenogenetic sires (CP), parthenogenetic dams + control sires (PC), and parthenogenetic dams + parthenogenetic sires (PP). For all progeny, a dam line main effect revealed that the parthenogenetic line dams had heavier offspring hatch weight and 4 wk body weight as well as higher 1st wk chick mortality versus control line dams. However, control line dams had the highest 4th wk chick mortality versus parthenogenetic line dams. In female virgin progeny, a dam by sire interaction revealed that PP, PC, and CP had the heaviest 1st egg in the clutch position versus CC. Also, eggs from PP had the highest number of eggs and the most female progeny exhibiting parthenogenesis versus CC. There was a linear increase in egg weight as clutch position increased for progeny from PP and CC yet a linear decline for CP. In conclusion, it appears that both the dam and sire selected for parthenogenesis impact progeny performance as parthenogenetic dams and sires additively contribute to the degree of parthenogenesis exhibited by virgin female progeny. Moreover, because parthenogenesis is known to exist in the modern poultry industry, even the accidental selection of the parthenogenetic trait in either males or females could have a negative impact on overall chick production and performance.

Parthenogenesis in birds: a review.

Parthenogenesis or 'virgin birth' is embryonic development in unfertilized eggs. It is a routine means of reproduction in many invertebrates. However, even though parthenogenesis occurs naturally in even more advanced vertebrates, like birds, it is mostly abortive in nature. In fact, multiple limiting factors, such as delayed and unorganized development as well as unfavorable conditions developing within the unfertilized egg upon incubation, are associated with termination of progressive development of parthenogenetic embryos. In birds, diploid parthenogenesis is automictic and facultative producing only males. However, the mechanisms controlling parthenogenesis in birds are not clearly elucidated. Additionally, it appears from even very recent research that these mechanisms may hinder the normal fertilization process and subsequent embryonic development. For instance, virgin quail and turkey hens exhibiting parthenogenesis have reduced reproductive performance following mating. Also, genetic selection and environmental factors, such as live virus vaccinations, are known to trigger the process of parthenogenesis in birds. Therefore, parthenogenesis has a plausible negative impact on the poultry industry. Hence, a better understanding of parthenogenesis and the mechanisms that control it could benefit commercial poultry production. In this context, the aim of this review is to provide a complete overview of the process of parthenogenesis in birds.

Investigating commercial in ovo technology as a strategy for introducing probiotic bacteria to broiler embryos.

Probiotics can improve broiler performance and reduce pathogens. Because the hatchery can be a source of contamination, delivering probiotics to the embryo before hatch is desirable. To date, probiotics have primarily been injected into eggs manually. Therefore, the objective of this study was to deliver various probiotic bacteria into broiler hatching eggs using an automated commercial in ovo injection system to evaluate hatchability of fertile eggs (HF). Three separate experiments were conducted using Lactobacillus acidophilus, Bacillus subtilis, or Bifidobacterium animalis. In each experiment, 7 treatments (non-injected control; dry punch control; diluent-injected control; and injections of 103 cfu, 104 cfu, 105 cfu, or 106 cfu of bacteria/50 µL of diluent) were evaluated using 10 replicates per treatment. For each experiment, 2,490 eggs were obtained from a commercial hatchery. Eggs were incubated under standard incubation conditions. At 10 d of incubation (doi), eggs were candled, and infertile eggs were removed. On 18 doi, all eggs were injected with the appropriate treatment using an automated in ovo injection system. Once all eggs were injected, they were transferred to hatching baskets and placed into the hatcher. On 21 doi, chicks were removed from the hatcher, counted, and weighed. Hatch residue analysis was conducted to determine infertile, early dead, mid dead, late dead, pipped, cracked, contaminated, and cull chick statuses of all unhatched eggs. Injecting L. acidophilus, even at a concentration as high as 106 cfu/50 µL, did not impact hatch residue analysis (P > 0.05). However, HF was significantly less for eggs treated with B. subtilis than for control eggs (P < 0.0001). For the non-injected control, HF was 91%, but as concentration of B. subtilis increased, HF decreased to as low as 1.67% for the 105 cfu treatment. Late deads, pipped, and contaminated egg percentages were higher, and chick BW was lower for B. subtilis treatment groups compared to controls. In conclusion, L. acidophilus and B. animalis but not B. subtilis, appear to be suitable candidates for in ovo injection as probiotics.

Tuberculosis trends in California correctional facilities, 1993-2013.

BACKGROUND: Incarcerated persons are disproportionately diagnosed with tuberculosis (TB). California has the second highest inmate population in the United States, but reports the highest number of cases. OBJECTIVE: To describe the TB epidemiology among incarcerated patients in California. METHODS: Trends in incidence were assessed using Poisson regression, and trends in percentage were assessed using weighted linear regression. Demographic and clinical characteristics were compared using χ2 or Mann-Whitney U tests. RESULTS: During 1993-2013, of the 64 090 TB cases reported, 2323 (4%) were correctional facility residents. Incidence in correctional facilities decreased until 2006 (annual per cent change [APC] -12.3%, 95%CI -14.4 to -10.1), but has since stabilized (APC 4.4%, 95%CI -2.1 to 11.4). Compared with state prisoners, federal prisoners were more likely to be male (98%, P = 0.03), persons arriving in the United States within 5 years of diagnosis (62%, P < 0.001), and born in Mexico (88%, P = 0.02), whereas local jail inmates were more likely to have a history of substance use (75%, P < 0.001) and homelessness (35%, P < 0.001). CONCLUSIONS: TB incidence in correctional facilities had steadily declined over the last two decades, but has recently leveled out. To promote further reduction in incidence among diverse incarcerated populations, health departments and correctional facilities should strengthen collaboration by conducting TB risk-based assessments.

Effects of embryo temperature estimation methodology on the determination of eggshell conductance values in Ross 708 broiler hatching eggs with consideration given to eggshell pigmentation variation.

In a preliminary experiment, the influence of pigment color variation on the temperature readings of the shells (EST) of live embryonated Ross 708 broiler hatching eggs was tested. Prior to set, eggshell color in L*, a*, and b* coordinates were measured at the equator of each egg. Eggshell pigmentation was found not to influence EST from zero through 18 d of incubation (DOI). In a subsequent experiment, EST, as well as air cell (ACT) and cloaca (CLT) temperature measurements were used for the calculation of absolute (GH2O) and specific (gH20) eggshell conductance values for these same types of eggs. An infrared thermometer was used to determine EST from zero to 19 DOI, ACT was measured using a transponder from 12 to 19 DOI, and CLT was determined using a transponder at 4:00 PM at 19 DOI. In the 12 to 19 DOI interval, the values for GH2O as well as for gH20 that were calculated using either EST or ACT were significantly correlated (r ≥ 0.99; P < 0.0001). A similar correlation level for both GH2O and gH20 was likewise observed in the 10:00 AM to 10:00 PM time period at 19 DOI when either EST, ACT, or CLT was used. However, in the 12 to 19 DOI interval, calculated GH2O and gH20 values based on ACT were significantly different from those based on EST. In addition, a significant difference in calculated GH2O and gH20 values resulted when ACT was used rather than when EST or CLT was used in the 10:00 AM to 10:00 PM time period at 19 DOI. In both time periods, GH2O and gH20 values calculated using ACT were significantly lower than those derived using the other 2 types of measurements. These findings suggest that although EST was not affected by shell coloration, because ACT more closely reflects embryo body temperature, it should be used to more accurately calculate the GH2O and gH20 of Ross 708 broiler hatching eggs.

Parental sex effect of parthenogenesis on hatchability and sperm-egg penetration in mated Chinese Painted quail (Coturnix chinensis).

Selecting quail for an increased incidence of parthenogenesis also impacts egg weight and albumen pH as well as reduces hatchability and fertility due to decreased sperm-egg penetration (SEP). However, it is unknown which parental sex is responsible for these changes in quail selected for parthenogenesis. Therefore, the objective of this study was to determine which sex influences egg weight, albumen pH, hatchability, and SEP in birds selected for parthenogenesis. In this study, 2 lines of birds were used: 1 line that was selected for parthenogenesis and 1 line not selected for parthenogenesis (control). Treatments were as follows: control females w/control males, control females w/parthenogenetic line males, parthenogenetic line females w/control males, and parthenogenetic line females w/parthenogenetic line males. Fresh eggs were collected daily, labeled and analyzed for albumen pH and SEP or incubated at 37.5 °C for 20 d of incubation. Eggs were candled at 10 days of incubation (DOI) and eggs exhibiting little or no embryonic development were removed and broken open to determine hatching failure. This was repeated at 20 DOI for eggs that did not hatch. A dam main effect for egg set weight existed with parthenogenetic line dams exhibiting heavier eggs than control dams. The parthenogenetic line dams and sires exhibited lower albumen pH and hatch but a higher incidence of parthenogenesis than control line dams or sires. However, only a sire main effect existed for fertility and SEP. Sires from the parthenogenetic line yielded the highest infertility due to lower SEP. In conclusion, both the parthenogenetic line dams and sires contribute to reduced reproductive performance. However, it appears that the sire from the parthenogenetic line is responsible for lower fertility due to a reduction in SEP. Because the sire has a negative impact on overall fertility, it is possible that males selected for parthenogenesis have poorer semen quality resulting in fewer sperm traversing the oviduct or penetrating the perivitelline layer.

Parthenogenesis in mated Chinese Painted quail (Coturnix chinensis) hens decreases sperm-egg penetration and alters albumen characteristics.

Parthenogenesis, embryonic development without fertilization, resembles very early embryonic mortality in fertilized eggs. Also, parthenogenesis alters egg albumen characteristics in virgin Chinese Painted quail hens genetically selected for parthenogenesis (PV). When these PV hens are mated (PM), hatchability is reduced versus control mated (CM) hens that were not genetically selected for parthenogenesis. However, it is unclear if parthenogenesis, which occurs in PM hens, reduces hatchability due to infertility and altered albumen characteristics. Sperm-egg penetration (SEP) holes are indicative of true fertilization and may be useful in identifying if eggs from PM hens exhibit a decrease in fertility versus CM hens. Therefore, the objectives of this study were to determine if parthenogenesis in PM hens (1) decreases SEP, (2) alters albumen characteristics similar to parthenogenesis in eggs from PV hens, and (3) yields albumen characteristics similar to fertilized eggs containing early mortality. Daily, PV and PM eggs were collected, labeled, and incubated for 10 days, then broken out to determine the incidence of parthenogenesis and albumen characteristics. Also daily, fresh PM and CM quail eggs were macroscopically examined to determine if an egg was infertile with no embryonic development, parthenogenetic, or fertile. Each of these eggs was then microscopically examined for SEP. For both PV and PM incubated eggs, parthenogenesis decreased albumen pH, O2, and protein concentrations yet increased Ca(2+) and CO2 concentrations versus eggs with no development. For incubated PM eggs, albumen pH and O2 were lower, yet CO2 was higher for eggs containing parthenogens or early dead embryos versus infertile eggs. For SEP, fresh eggs classified as infertile or parthenogenetic from PM and CM hens had similar SEP holes but only one sixth as many SEP holes as eggs classified as fertilized. Eggs from CM hens had 3.5 times as many SEP holes as PM eggs. In conclusion, parthenogenesis that occurs in mated quail hens inhibits fertility and alters albumen characteristics similarly to parthenogenesis in unfertilized eggs and early embryonic mortality in fertilized eggs.

Influence of 6 different intestinal bacteria on Beltsville Small White turkey semen.

The turkey industry relies totally on artificial insemination to continue and improve production. If something compromises the insemination process, such as contaminated semen, it becomes a detrimental loss to the industry. Bacteria have been found in broiler breeder males to reduce sperm motility. The Sperm Quality Index (SQI) is a quick method to determine avian sperm motility using the sperm quality analyzer (SQA). Therefore, the objective of this study was to determine if bacteria have an effect on turkey sperm motility using the SQA. For the experiment, one mL of pooled neat semen was collected from Beltsville Small White Turkey toms. Six intestinal bacteria, Bifidobacterium animalis, Campylobacter jejuni, Clostridium bifermentans, Escherichia coli, Lactobacillus acidophilus, and Salmonella enterica were grown overnight. For each bacterium, 4 treatments were made that consisted of exposing pooled semen to either saline, sterile broth, an overnight culture of each individual bacterium, or a centrifuged pellet of each bacterium re-suspended in saline. The experiment was repeated 3 times. Once the semen was exposed to the respective treatment, a portion was pulled into a capillary tube and placed into the SQA to obtain the SQI. Each treatment was evaluated at zero, 10, and 20 min creating a completely randomized design with a split plot over time. A pH reading also was taken at each time point. The results indicated that all broths containing bacteria immediately reduced turkey sperm motility. Sperm became practically immotile in overnight cultures of Clostridium, Bifidobacterium, or Lactobacillus However, there was a time by treatment interaction in the SQI for Campylobacter, Clostridium, E. coli, Salmonella, and Lactobacillus The pH of semen decreased upon exposure to Bifidobacterium and Lactobacillus No difference in pH was found when semen was exposed to E. coli, Campylobacter, Salmonella, or Clostridium treatments. In conclusion, the results reveal when turkey semen is exposed to different bacteria, sperm motility is immediately reduced, which could be possible from bacterial attachment or bacterial by-products providing an undesirable environment for sperm.

Impact of oral Lactobacillus acidophilus gavage on rooster seminal and cloacal Lactobacilli concentrations.

The use of antibiotics in poultry is being heavily scrutinized, therefore alternatives such as probiotics are being investigated. Lactobacilli spp. are a commonly used bacteria in formulating probiotics, and the addition of Lactobacilli to broiler diets has demonstrated increased growth rates, stimulated immune systems, and reduced pathogen loads in the gastro-intestinal tract ( GI: ) tract. However, previous research has shown that when rooster semen is directly exposed to Lactobacillus acidophilus (L. acidophilus) sperm quality is reduced. Therefore, the objective of the current study was to determine if oral administration of L. acidophilus increases the concentration of Lactobacilli in semen as well as the cloaca. A total of 30 roosters were used: 15 roosters were gavaged with 1X PBS (Control) and 15 roosters were gavaged with 10(7) cfu/mL of L. acidophilus (Treated). All roosters were gavaged for 14 consecutive days. Semen was collected on a 3 d interval, and cloacal swabs were collected on a 2 d interval, beginning on the first day prior to oral administration. Semen and cloacal swabs were serial diluted, and 100 µL of each dilution was then plated on Man, Rogosa, Sharpe ( MRS: ) agar plates. All plates were incubated for 48 h at 37°C under anaerobic conditions and counted. All Lactobacilli counts were first log transformed, then log transformed (day 0) pre-counts were subtracted from the log transformed day counts providing log differences for the analysis. Seminal Lactobacilli counts were not altered by treatments. However, the main effect of treatment (P = 0.026) for cloacal counts indicated that roosters gavaged with Lactobacilli yielded higher counts than the controls. Additionally, cloaca samples also demonstrated a treatment by day interaction trend (P = 0.082), where Lactobacilli was higher in the L. acidophilus gavaged roosters than the controls only on days 3, 5, 13, and 15. In conclusion, the addition of L. acidophilus to the male breeder diet over extended periods may increase concentrations of Lactobacilli in the cloaca even higher than the concentrations observed in this study. If Lactobacilli reaches high enough concentrations in the cloaca, then sperm quality may be impacted which could lead to poor fertility within the breeder flock.

Parthenogenetic embryos from unfertilized Chinese painted quail eggs alter albumen pH, gases, and ion concentrations during incubation.

Parthenogenesis is a form of embryonic development that occurs without fertilization. Recently, parthenogenesis has been reported in Chinese painted quail eggs. In Japanese quail, it has been shown that albumen pH of incubated fertile eggs is lower than that of incubated infertile eggs. However, it is unknown if alterations, similar to those in incubated fertile eggs, occur in albumen pH, gases, or ion concentrations from unfertilized eggs exhibiting parthenogenetic development. Therefore, the objective of this study was to determine if any differences in pH, gases, or ion concentrations exist between incubated unfertilized eggs exhibiting parthenogenetic development versus unfertilized eggs with no development over incubation. In this study, eggs were collected daily from Chinese painted quail hens that were separated from males at 4 weeks of age, before sexual maturity. Eggs were stored for 0 to 3 days at 20 °C and incubated at 37.5 °C for 12 days. Eggs were weighed before and after incubation to obtain percentage egg weight loss. After incubation, embryo size and albumen O2, CO2, Ca(2+), Na(+), and Cl(-) concentrations as well as pH were obtained from each incubated egg. Over incubation, albumen from unfertilized eggs exhibiting parthenogenetic development had a lower pH as well as less O2 and Cl(-), yet a higher Ca(2+) and Na(+) concentration as compared with the albumen of unfertilized eggs with no development. Also, eggs exhibiting parthenogenetic development had a higher albumen CO2 concentration as compared with eggs without development. The rate of egg weight loss was much lower in eggs exhibiting parthenogenetic development as compared with eggs without development. Also, as parthenogen size increased, there was a decrease in albumen pH, O2, and Cl(-), yet an increase in CO2 and Ca(2+). In conclusion, it appears that, over incubation, parthenogenetic development from unfertilized eggs alters the composition of albumen as compared with the albumen from unfertilized eggs with no parthenogenetic development.

Selection for the parthenogenetic trait in Chinese Painted Quail (Coturnix chinensis) affects hatchability parameters.

Poultry are capable of laying unfertilized eggs in which embryonic development occurs; this phenomenon is called parthenogenesis. Through genetic selection, the incidence of parthenogenesis in virgin Chinese Painted Quail hens can be increased. However, it is unknown if selection for this trait affects hatchability of fertilized eggs. Therefore, the objective of this study was to determine if genetic selection for parthenogenesis in virgin Chinese Painted Quail affects hatchability of mated hens. At 4 wk of age, females were separated from males and individually caged. Daily, eggs were collected, labeled, and stored from 0 to 3 d at 20°C then incubated at 37.5°C. At d 10 of incubation, eggs from virgin hens were broken out to determine the incidence of parthenogenesis. After the incidence of parthenogenesis in virgin hens was determined, hens were selected for mating. Eggs from the parent stock and generations 1 through 7 that did not hatch were examined for stage of hatching failure and categorized as infertile or possible parthenogens as well as early, middle, and late embryonic mortality. For hatch of set and hatch of fertile eggs, generation 1 hatched the greatest percentage of eggs compared with the other generations. Also, as the generation of selection increased, there was a linear decline in the percentage of eggs hatched for both hatch of set and hatch of fertile eggs. Additionally, generation 2 had the highest percentage of infertile eggs. There was a linear increase as generation of selection increased for both the percentage of eggs exhibiting possible parthenogens and early embryonic mortality. Unlike early embryonic mortality, the parent generation had the greatest percentage of embryos die in both the middle and late stage of incubation. In conclusion, it appears that selecting Chinese Painted Quail for the parthenogenetic trait impairs hatchability when hens are mated.

Impact of 6 different intestinal bacteria on broiler breeder sperm motility in vitro.

Male fertility is often evaluated by measuring sperm parameters, including concentration, viability, and motility. This is important because after copulation occurs, sperm must overcome many barriers in the female reproductive tract to fertilize the ovum. In mammalian species, sperm have been shown to have reduced motility when bacteria are present. In male broiler breeders, bacteria have been associated with spermatozoa, but their effect on motility has not been investigated. The sperm quality index is a modern rapid method of evaluating avian sperm motility. Therefore, the objective of this study was to use the sperm quality index to determine if broiler breeder sperm motility is reduced when semen is exposed to various bacteria. In this experiment, semen was collected from 20 broiler breeders to obtain a pooled neat semen sample. Six different intestinal bacteria, Salmonella enterica, Escherichia coli, Campylobacter jejuni, Clostridium bifermentans, Lactobacillus acidophilus, and Bifidobacterium animalis were cultured overnight. For each bacterium, 50 µL of semen was diluted in 450 µL of saline, sterile broth, or the overnight culture, creating 3 treatments. The experiment was repeated twice. In each treatment, 3 replicates were evaluated at 0 and 10 min postinoculation, creating a completely randomized design with a split plot over time. Also, the pH was measured for each treatment at 0 and 10 min. The results indicated that all broths containing bacteria immediately reduced broiler breeder sperm motility when compared with the controls (P < 0.0001), but broths containing Bifidobacterium or Lactobacillus virtually made sperm immotile. Although broth containing Salmonella, Campylobacter, and Bifidobacterium immediately reduced sperm motility, the reduction did not change over time. Broths containing E. coli, Clostridium, and Lactobacillus reduced sperm motility immediately, but over time motility continued to decrease. However, pH was increased when semen was exposed to the E. coli and Campylobacter treatment, but when semen was exposed to Bifidobacterium and Lactobacillus treatments, pH was reduced. In conclusion, the results indicate that bacteria can reduce broiler breeder sperm motility upon exposure.

Metabolic and hormonal responses of growing modern meat-type chickens to fasting.

1. The present study compared the effects of fasting on circulating concentrations of glucose, insulin and glucagon in male and female modern meat-type chickens (Ross 708) at three ages (19 d, 33 d and 47 d). 2. Plasma concentrations of glucose were reduced by fasting with reductions of 24.9% (19-d-old), 22.6% (33-d-old) and 17.9% (47-d-old) in broiler chickens fasted for 12 h. 3. Plasma concentrations of insulin decreased with fasting. For instance, circulating concentrations of insulin declined after 6 h of fasting by 45.7%, 54.7% and 50.0%, respectively, in 19-d-old, 33-d-old and 47-d-old broiler chickens. 4. Plasma concentrations of glucagon were increased by fasting. Plasma concentrations of glucagon were elevated by 3.79% (19-d-old), 3.51% (33-d-old) and 3.79% (47-d-old) with 6 h of fasting and remained elevated with 12 h, 18 h and 24 h of fasting.

Evaluation of carryover effects of dietary lysine intake by Cobb 500 broiler breeder hens.

Two experiments were conducted to examine the progeny performance of broiler breeder (BB) hens fed diets differing in digestible lysine (dLys). A total of 240 Cobb 500 BB pullets and 40 cockerels, 20 wk of age, were used for each of 2 experiments. In experiment 1, treatment diets were fed from 35 to 45 wk of age. Treatment 1 and 2 diets were formulated with common feed ingredients and BB had dLys daily intakes of 1,200 (IDL-1,200) and 1,010 mg/hen per d (ID-1,010), respectively. Treatments 3 and 4 were composed of semipurified diets formulated to dLys intakes of 1,010 (SPL-1,010) and 600 mg/hen per d (SP-600), respectively. In experiment 1, chick weight at hatch was lower (P < 0.05) for those that came from the SP-600 and SPL-1,010-fed hens, but 42 and 56 d BW was similar for all treatments. Improvements (P < 0.10) in feed conversion ratio were seen at 42 and 56 d for chicks from ID-1,010-fed hens compared with IDL-1,200 hens. For experiment 2, diets were fed to BB hens from 24 to 42 wk of age. Treatment 1 was composed of corn-soybean meal and formulated to a dLys intake of 1,000 mg/hen per d (CS-1,000). Treatments 2, 3, and 4 had inclusion of 25% distillers dried grains with solubles (DDGS) with dLys intake levels of 1,000 (DDGS-1,000), 800 (DDGS-800), and 600 (DDGS-600) mg/hen per d, respectively. Progeny performance was evaluated from eggs collected at wk 26, 31, and 36. Chick hatch weight was similar for all 3 hatches. Chicks from 26-wk-old BB hens and fed DDGS-600 diets resulted in lower (P < 0.05) BW, carcass weight, and breast weight, and higher (P < 0.05) back half (drumsticks and thighs) yield, at 42 d of age. Growout studies performed on eggs laid during wk 31 and 36 revealed no effect on growth or carcass parameters. In conclusion, the results from experiments 1 and 2 may not be comparable, and therefore, the impact on progeny performance needs to be further evaluated.

The relationship of parthenogenesis in virgin Chinese Painted quail (Coturnix chinensis) hens with embryonic mortality and hatchability following mating.

Unfertilized chicken, turkey, and quail eggs are capable of developing embryos by parthenogenesis. However, it is unknown if the physiological mechanisms regulating parthenogenesis in virgin hens may actually work against fertilization, embryonic development, and hatchability of eggs from these same hens following mating. Additionally, because most parthenogenic development closely resembles early embryonic mortality in fertilized eggs during the first 2 to 3 d of incubation, it is possible that many unhatched eggs classified as containing early embryonic mortality may actually be unfertilized eggs that contain parthenogens. Therefore, the objective of this study was to examine the relationship of parthenogenesis before mating with embryonic development and hatchability characteristics after mating. Based upon their ability to produce unfertilized eggs that contain parthenogens, 372 virgin Chinese Painted quail hens were divided into 7 groups, according to their incidence of parthenogenesis: 0, 10, 20, 30, 40, 50, and greater than 50% parthenogenesis. Males were then placed with these hens so that fertility, embryonic mortality, and hatchability could be evaluated for each hen. Hatchability of eggs set, hatchability of fertile eggs, and late embryonic mortality declined dramatically as the incidence of parthenogenesis increased. On the other hand, early embryonic mortality increased as parthenogenesis increased. Fertility was not different across the 7 parthenogenesis hen groups, perhaps because unfertilized eggs that exhibited parthenogenesis resembled and were therefore classified as early embryonic mortality. In conclusion, virgin quail hens that exhibit parthenogenesis appear to have impaired embryonic development and hatchability following mating. Additional sperm-egg interaction and embryonic research is needed to determine if a large portion of the early embryonic mortality experienced by mated hens that exhibit parthenogenesis as virgin hens is in fact embryonic development in unfertilized eggs.