Embryo Mortality Research Articles

Effect of in ovo injection of DL-methionine on hatchability, embryo mortality, hatching weight, blood biochemical parameters and gastrointestinal tract development of breeder chicks

The current investigation was conducted to test the potential effects of in ovo feeding of DL-methionine (MET) on hatchability, embryonic mortality, hatching weight, blood biochemical parameters and development of heart and gastrointestinal (GIT) of breeder chick embryos. 224 Rhode Island Red fertile eggs were randomly distributed into seven experimental treatments: untreated egg (control), buffered saline (0.5% NaCl), and five solutions containing increased levels of MET (0.5, 1.0, 1.5, 2.0 and 2.5%) + 0.5% NaCl, being separated into four groups/replicates (each one with 8 eggs), totaling 32 eggs/treatment. All embryos submitted to in ovo injection with MET presented a decrease in the hatchability results and an increase in the results of intermediary embryonic mortality. Chicks hatched from eggs injected with until to 1.0% MET were heavier and presented better development of the heart and GIT, especially important organs and regions for digestion and nutrient absorption. Conclusively, the in ovo feeding using MET showed positive impacts on hatching weight and GIT development of breeder chicks. However, caused negative impacts on hatchability when used at high levels.

Homocysteine-Potential Novel Diagnostic Indicator of Health and Disease in Horses.

Homocysteine is an endogenous, non-protein sulfuric amino acid, an intermediate metabolite formed by the methionine transmethylation reaction. Its elevated serum concentration in humans, hyperhomocysteinemia, is a sensitive indicator and a risk factor for coagulation disorders, cardiovascular diseases and dementia. However, the role of homocysteine in veterinary species has not been unequivocally established. Although some research has been conducted in dogs, cats, cattle and pigs, relatively few studies on homocysteine have been conducted in horses. So far, it has been established in this species that homocysteine has an atherogenic effect, plays a role in early embryo mortality and is responsible for the induction of oxidative stress. These preliminary findings support establishing a reference range in a normal population of horses, including horses in training and merit further investigations into the role of this amino acid in health and disease in this species.

Late embryo mortality in Holstein cows inseminated with Holstein or Limousine frozen semen.

The objective of this study was to compare the late embryo mortality (LEM) rate (losses approximately between 32 and 53 days of gestation) and Pregnancy Specific Protein B (PSPB) and progesterone (P4) concentrations on day 32 post AI in Holstein cows bred with either Holstein or Limousine semen. A sample size of 1082 cows per group diagnosed pregnant between 28- and 35-days post breeding was calculated. The study consisted of evaluating LEM (%) in a cohort of Holstein cows bred with Holstein semen (HO × HO) or Limousine semen (HO × LM), to compare pregnancy loss from 28 to 35 days post breeding to 50-57 days post breeding. A logistic regression model to compare embryo losses was developed considering as main explanatory variable the cohort (HO × HO embryo vs. HO × LM embryo), correcting by lactation number, breeding season, days to breeding and AI technician. HO × HO embryos had greater LEM (15.16%) than HO × LM embryos (9.79%). Cows bred in summertime had higher LEM (15.23%) than cows bred in no-summertime (9.88%). There were no differences among AI technicians. Within summertime there was no difference in LEM (%) between groups within each lactation number; yet, within no-summertime, LEM (%) was higher in HO × HO than HO × LM within each lactation number. Pregnancy SPB optical densities were significantly greater in the HO × HO than in the HO × LM (p= .023) group; yet, the concentration of P4 was not different between groups (p> .05).

Impacts of Deoxygenation and Hypoxia on Shark Embryos Anti-Predator Behavior and Oxidative Stress.

Climate change is leading to the loss of oxygen content in the oceans and endangering the survival of many marine species. Due to sea surface temperature warming and changing circulation, the ocean has become more stratified and is consequently losing its oxygen content. Oviparous elasmobranchs are particularly vulnerable as they lay their eggs in coastal and shallow areas, where they experience significant oscillations in oxygen levels. Here, we investigated the effects of deoxygenation (93% air saturation) and hypoxia (26% air saturation) during a short-term period (six days) on the anti-predator avoidance behavior and physiology (oxidative stress) of small-spotted catshark (Scyliorhinus canicula) embryos. Their survival rate decreased to 88% and 56% under deoxygenation and hypoxia, respectively. The tail beat rates were significantly enhanced in the embryos under hypoxia compared to those exposed to deoxygenation and control conditions, and the freeze response duration showed a significant opposite trend. Yet, at the physiological level, through the analyses of key biomarkers (SOD, CAT, GPx, and GST activities as well as HSP70, Ubiquitin, and MDA levels), we found no evidence of increased oxidative stress and cell damage under hypoxia. Thus, the present findings show that the projected end-of-the-century deoxygenation levels elicit neglectable biological effects on shark embryos. On the other hand, hypoxia causes a high embryo mortality rate. Additionally, hypoxia makes embryos more vulnerable to predators, because the increased tail beat frequency will enhance the release of chemical and physical cues that can be detected by predators. The shortening of the shark freeze response under hypoxia also makes the embryos more prone to predation.

The Effect of In Ovo Administration of Rosemary Essential Oil on Hatchability, Relative Hatching Weight, and Embryo Mortality Rate in Japanese Quail (Coturnix coturnix japonica).

This study aimed to determine the effects of air sac injection of rosemary essential oil at different concentrations in ovo in quail eggs on hatching rate, relative chick weight at hatching, and embryonic mortality rate. A total of 1060 Japanese quail eggs were divided into four groups: negative control (non-injected), positive control (30 µL sterile distilled water/egg), and two treated groups with 1 and 3 µL oil/egg, respectively. The concentration of 3 µL/egg showed a toxic effect on embryonic development, as revealed by the significantly (p = 0.015) higher post-injection mortality rate (18.21%) compared to 1 µL/egg with 8.3%. Furthermore, hatchability was significantly increased (p = 0.0001) with 1 µL/egg compared to 3 µL/egg with 69.1% and 44.48%, respectively. No significant difference was observed between the concentration of 1 µL/egg and the control groups (p = 0.822). Both l and 3 µL essential oil/egg significantly enhanced (p = 0.0001) relative chick weight at hatching by 67.14% and 70.32%, respectively, compared to the control groups. In conclusion, injecting eggs with 1 µL oil/egg showed positive effects both on hatching and relative chick weight. The concentration of 3 µL/egg was revealed to be toxic, with dramatic effects on embryonic survival.

The effect of storage periods and SPIDES on embryonic mortality, hatching characteristics, and quality of newly hatched chicks in broiler eggs

sEgg storage duration can affect embryo mortality, hatching characteristics, hatching time, and post-hatch chick quality. In order to assess these effects, the impact of storage duration (5 days, 10 days, 15 days) and short incubation period during egg storage (SPIDES) investigated further 18, 900 eggs of broiler breeder (ROSS 308) in 3 × 2 factorial arrangement design. In the SPIDES treatment, the egg shell temperature was raised from its storage temperature (18 °C) and held at 100 °F for 3.5 h. Storage periods could significantly (P < 0.05) influence on embryo mortality (total, early, middle, and late), hatchability of both the total eggs and fertile eggs. The SPIDES treatment had a significant (P < 0.05) impact on a lower embryonic death rate and improved egg hatchability. Eggs stored for 5 days and eggs treated with SPIDES significantly (P < 0.001) shorten hatching time, batch’s 90% hatching time (T 90% H), mean hatching time (MHT), maximal hatching period (MHP), and hatching window (HW). Chick quality was also determined, whereas storing eggs for 5 days and using the SPIDES treatment resulted in enhanced (P < 0.001) chick weight relative to egg weight (CW/EW), activity (AC), and chick quality score (CQS). The residual yolk sac weight (RYSW), unhealed navel (UHN %), and dirty feather (DF%) recorded the lowest (P < 0.001) values compared to long storage periods and control group. Finally, stored for 5 days treated by SPIDES positively affected the hatchability characteristics, the shortening hatching time, and the quality of chicks. Regarding the results, it was confirmed that using the SPIDES treatment to prevent the harmful effects of broiler eggs being stored for an extended period of time is a viable option.

Toxic Effects Associated With Neonicotinoid Exposure on Non-target Organisms: A Review

The neonicotinoid class of insecticide is nicotine-like neuro-toxicants used to control the pests of agriculture crops and ornamental plants. They act as selective agonists of acetylcholine receptors in the central nervous system of insect pests and work by disrupting their nerve impulses. Some of the properties of this class of insecticides are a long half-life in soil, low volatility, and higher water solubility, leading to their accumulation in soil, underground water, and water bodies due to surface runoff. This, in turn, results in exposure to many beneficial non-target aquatic and soil fauna such as arthropods, fish, birds, mammals, etc. Although it has a selective mode of action for insects, some in vivo and in vitro investigations have also shown toxicity in non-target invertebrates and vertebrates. Initially, neonicotinoid toxicity was observed in honey bees, which are essential pollinators of crops. Later, studies reported the accumulation of neonicotinoid residues leading to the mortality of aquatic fauna, including salt marsh and freshwater mosquitoes, brine shrimp, fleas, and crayfish. Imidacloprid exposure led to disruption of larval development in the Mayfly larvae. Also, earthworms that play a crucial role in enhancing soil fertility were drastically affected by acetamiprid, clothianidin, imidacloprid, nitenpyram, and thiacloprid. Apart from these, toxicological impacts were also observed in vertebrates such as birds, where imidacloprid, clothianidin, acetamiprid, and thiacloprid caused reproductive, metabolic, and morphological alterations. Similarly, imidacloprid and acetamiprid caused gills, brain and liver dysfunction with embryo mortality. Even after the selective action of neonicotinoids, instances of mammalian toxicity were also reported in many in vivo studies. DNA damage and liver dysfunctions due to imidacloprid in rabbits were observed in various studies. In a recent study, imidacloprid exposure led to DNA damage and oxidative stress in bone marrow-derived mesenchymal cells of buffalo. Also, many instances of neurotoxicity, reproductive toxicity, immunotoxicity, genotoxicity and cytotoxicity in mouse and rat models were observed due to different neonicotinoids. Many in vitro studies using mammalian cell lines have also established potential risks of neonicotinoid exposure. This review, therefore, is a compilation of various toxicity studies of different types of neonicotinoid pesticides in both nontarget invertebrates and vertebrates, including several kinds of toxicities caused in mammals with neonicotinoid exposure.

Evaluation of the Grade of Hatching Eggs of Sensi-1 Agrinak and KUB Crossbred Chicken on Hatching and Production Performances

The potential of Sensi-1 Agrinak and KUB crossbred so-called SENKUB chicken as dual-purpose chicken can be increased by selecting hatching egg weights to get DOC with good quality. The study was conducted to evaluate the grades of hatching eggs of SENKUB chicken on hatching and production performances. The complete randomized design was applied in this experiment with 3 treatments. The treatments are the grades of hatching egg (small 36-40 g, medium 41-45 g, and large 46-50 g). The hatching process used two stages hatching machine. All DOCs were selected according to the Indonesia National Standard (SNI). The selected DOCs were raised in different pens to evaluate the production performances. The hatching showed that the large eggs have significantly the highest fertility, hatchability, hatching weight, and salable chick but the lowest weight loss. Embryo mortality was the same between different hatching egg grades. The best-weight DOC was obtained from medium and large hatching eggs. The hatching egg grades do not significantly affect the production performances of the hatched chicken. However, the chickens with small and large hatching eggs resulted in better production performance.

11.4 T ultra-high static magnetic field has no effect on morphology but induces upregulation of TNF signaling pathway based on transcriptome analysis in zebrafish embryos

As magnetic resonance imaging (MRI) scanners with ultra-high field (UHF) have optimal performance, scientists have been working to develop high-performance devices with strong magnetic fields to improve their diagnostic potential. However, whether an MRI scanner with UHF poses a risk to the safety of the organism require further evaluation. This study evaluated the effects of 11.4 Tesla (T) UHF on embryonic development using a zebrafish model. Multiple approaches, including morphological parameters, physiological behaviors, and analyses of the transcriptome at the molecular level, were determined during 5 days after laboratory-controlled exposure from 6 hour post fertilization (hpf) to 24 hpf. No significant effects were observed in embryo mortality, hatching rate, body length, Left-Right patterning, locomotor behavior, etc. RNA-sequencing analysis revealed up-regulated tumor necrosis factor (TNF) inflammatory factors and activated TNF signaling pathways in the 11.4 T exposure group. The results were further validated using qPCR. Our findings indicate that although UHF exposure under 11.4 T has no effect on the development of zebrafish embryos, it has specific effects on the immune response that require further investigation.

Toxicity of common biocides used in aquaculture to embryos and larvae of Brachymystax tsinlingensis Li.

Brachymystax tsinlingensis Li is a threatened fish species endemic to China. With the problems of environmental factors and seeding breeding diseases, it is important to further improve the efficiency of seeding breeding and the basis of resource protection. This study investigated the acute toxicity of copper, zinc and methylene blue (MB) on hatching, survival, morphology, heart rate (HR) and stress behaviour of B. tsinlingensis. Eggs (diameter: 3.86 ± 0.07 mm, weight: 0.032 ± 0.004 g) of B. tsinlingensis were selected randomly from artificial propagation and developed from eye-pigmentation-stage embryos to yolk-sac stage larvae (length: 12.40 ± 0.02 mm, weight: 0.03 ± 0.001 g) and exposed to different concentrations of Cu, Zn and MB for 144 h in a series of semi-static toxicity tests. The acute toxicity tests indicated that the 96-h median lethal concentration (LC50 ) values of the embryos and larvae were 1.71 and 0.22 mg l-1 for copper and 2.57 and 2.72 mg l-1 for zinc, respectively, whereas the MB LC50 after 144-h exposure for embryos and larvae were 67.88 and 17.81 mg l-1 , respectively. The safe concentrations of copper, zinc and MB were 0.17, 0.77 and 6.79 mg l-1 for embryos and 0.03, 0.03 and 1.78 mg l-1 for larvae, respectively. Copper, zinc and MB treatments with concentrations greater than 1.60, 2.00 and 60.00 mg l-1 , respectively, led to a significantly low hatching rate and significantly high embryo mortality (P < 0.05), and copper and MB treatments with concentrations greater than 0.2 and 20 mg l-1 led to significantly high larvae mortality (P < 0.05). Exposure to copper, zinc and MB resulted in developmental defects, including spinal curvature, tail deformity, vascular system anomalies and discolouration. Moreover, copper exposure significantly reduced the HR of larvae (P < 0.05). The embryos exhibited an obvious change in behaviour, converting from the normal behaviour of emerging from the membrane head first to emerging tail first, with probabilities of 34.82%, 14.81% and 49.07% under copper, zinc and MB treatments, respectively. The results demonstrated that the sensitivity of yolk-sac larvae to copper and MB was significantly higher than that of embryos (P < 0.05) and that B. tsinlingensis embryos or larvae might be more resistant to copper, zinc and MB than other members of the Salmonidae family, which benefits their resource protection and restoration.

Effect of photoperiod during incubation on embryonic temperature, hatch traits, and performance of 2 commercial broiler strains

Provision of light during incubation has shown the potential to enhance hatching traits and affect posthatch productivity, physiology, and behavior. In this study, 2 repeated trials were conducted to investigate the effect of photoperiod and strain on the embryo temperature, hatching traits and posthatch growth performance of 2 commercial strains of broilers (Ross 308 and Cobb 500). In each trial, hatching eggs were randomly distributed into 6 incubators with 3 photoperiod treatments: blue LED light for 12 h d-1 (12L:12D) or 18 h d-1 (18L:6D) during entire incubation were compared with no illumination condition (DARK). Data were analyzed as a 3×2 factorial arrangement with the trial as the blocking factor. Embryos incubated under 12L:12D and 18L:6D had lower air cell temperature (P < 0.05) than the DARK embryos from d 13 of incubation onward except on the day of candling. The response of air cell temperature to periodic illumination differed between 2 strains. Cobb embryos had lower air cell temperature in 12L:12D than those incubated with 18L:6D from d 16 of incubation onward, whereas lower air cell temperature was found in Ross embryos when illuminated with 18L:6D photoperiod compared to those under 12L:12D. The 12L:12D treatment was associated with improved (P < 0.05) navel closure condition of hatchlings. There were no differences in hatchability, embryo mortality, body weight, or length at hatch among photoperiod groups or its combination with strain. No differences in production parameters were found between DARK and illuminated groups. However, 12L:12D had heavier (P < 0.05) body weight on d 14 of age and higher (P < 0.05) body weight gain than 18L:6D from d 7 to 14 of age. The results of this study indicate that providing blue LED light up to 18 h d-1 has no detrimental effect on production of broilers, however, 12L:12D light regime improved chick quality at hatch compared to DARK and resulted in heavier birds by d 14 compared to 18L:6D.

Influence of Dietary Phytase Inclusion Rates on Yolk Inositol Concentration, Hatchability, Chick Quality, and Early Growth Performance

The aim of this study was to determine the influence of dietary phytase in breeder hens on yolk nutrients, hatchability, chick quality, and growth rate of their progeny, and their subsequent performance to 42 d post-hatch when fed diets with the same phytase concentrations. Breeder hens (n = 216) were divided into 3 groups receiving nutrient-adequate diets with reduced calcium (Ca) and phosphorus (P) (by 0.16% and 0.15%, respectively), supplemented with either 500, 1500, or 4500 FTU/kg phytase from 27 to 50 weeks of age. Eggs were collected at 38 weeks of age and incubated. On the day of hatch, the chick quality and hatchability were determined, and 18 chicks/group were euthanized for yolk sac collection and the determination of inositol and glycerol concentrations. The remaining chicks were divided into three groups, receiving different diets with reduced Ca and P (by 0.16% and 0.15%, respectively), supplemented with 0, 500, or 1500 FTU/kg phytase to 42 d post-hatch. Increasing the phytase concentration in the breeder hen diet linearly (p < 0.05) increased the number of early embryo deaths and decreased the number of late deaths and pips. The inositol concentration in the yolk sac at day of hatch increased (quadratic; p < 0.05) as the phytase dose increased in the breeder hen diet. The breeder hen diet (p < 0.05) influenced the body weight (BW), feed intake (FI), and feed conversion ratio (FCR) up to 21 days of age. The supplementation of breeder hen diets with 1500 FTU/kg phytase increased the concentration of sodium (Na), magnesium (Mg), potassium (K), manganese (Mn), and zinc (Zn) in the yolk sac. The inclusion of phytase doses up to 4500 FTU/kg appeared to influence embryo mortality, chick feed intake, and BW gain to 21 days and the FCR throughout the entire production phase.

Transcriptome analysis of high mortality phenomena in polyploid fish embryos: An allotriploid embryo case study in hybrid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂)

Triploidization of diploid fish is an ideal means for achieving polyploid breeding. During the preparation of allotriploid grouper (Epinephelus fuscoguttatus ♀ × Epinephelus lanceolatus ♂), the allotriploid embryos (T) had a much higher morality from the high blastula to middle gastrula stage compared to the normal hybrid diploids (D). Therefore, the internal regulation of gene expression in allotriploid fertilized grouper eggs was explored by comparing the transcriptional profiles of T and D mapped at the high blastula (1), middle gastrula (2), and crystal (3) stages using the Illumina RNA-seq technology. There were 2357 differentially expressed genes (DEGs) found only in T1_vs_T2 and 1585 DEGs found only in T2_vs_T3. According to the GO functional enrichment analysis, when T developed from the high blastula to middle gastrula stage (i.e., pre-embryonic development; T1_vs_T2), the DEGs showed significant enrichment in terms associated with the cell cycle and cytoskeleton, indicating that cell proliferation was inhibited. KEGG pathway analysis of DEGs showed significant enrichment in mRNA processing and translation and in pathways associated with programmed cell death (PCD). Cell differentiation can trigger PCD and the combined effects of up-regulated ripk1, dnm1l, pgam5, Slc25a5, and vdac1 expression and down-regulated fadd expression during pre-embryonic development may have been the main cause of mass mortality in embryos up until the middle gastrula stage. A two-by-two comparison of D2_vs_T2 DEGs at the middle gastrula stage to the aforementioned DEGs expressed only in T revealed that the Parp4 and pole were down-regulated in the pre-development of T, which might lead to decreased DNA repair efficiency. These results could have reflected either (1) the altered physiological state of the cells due to cold-shock treatment or (2) the sensitivity of pre-embryonic development to an elevated number of chromosomes, which induced disrupted gene expression and abnormal cell division. In conclusion, the above study helps explain why allotriploid grouper exhibit high mortality at the middle gastrula stage, improves our understanding of the regulation of triploid embryo development, and provides basic information for reducing mortality at the middle gastrula stage.

NDUFA8 potentially rescues Wolbachia-induced cytoplasmic incompatibility in Laodelphax striatellus.

The endosymbiont Wolbachia manipulates host reproduction by several strategies, one of the most important of which is cytoplasmic incompatibility (CI). CI can be rescued when Wolbachia-infected males mate with females infected with the same Wolbachia strain. However, the potential rescue mechanism of CI in the small brown planthopper Laodelphax striatellus is unclear. In this study, comparative transcriptome analysis was applied to explore the effect of Wolbachia on L. striatellus eggs. A total of 1387 differentially expressed genes were identified. RNA interference of 7 Wolbachia-upregulated key planthopper genes reduced egg reproduction, suggesting that Wolbachia might improve fecundity in L. striatellus by affecting these 7 genes. Suppressing the expression of another upregulated gene, NDUFA8 (encoding NADH dehydrogenase [ubiquinone] 1 α subcomplex subunit 8-like) by RNA interference significantly increased the mortality of early embryos without affecting the number of deposited eggs. Wolbachia infection upregulated the mRNA level of NDUFA8, and dsNDUFA8 treatment of Wolbachia-infected females recreated CI-like symptoms, suggesting that NDUFA8 is associated with the rescue phenotype. Because all L. striatellus populations worldwide are infected with Wolbachia, NDUFA8 is a potential pest control target.

Aerial Application of Organic Pellets Eliminates Lake Trout Recruitment from a Primary Spawning Reef in Yellowstone Lake

AbstractInvasive Lake Trout Salvelinus namaycush in the Yellowstone Lake ecosystem have been gillnetted since 1995 to suppress the population and allow for recovery of native Yellowstone Cutthroat Trout Oncorhynchus clarkii bouvieri. Although gillnetting is effective (Lake Trout population growth rate λ ≤ 0.6 during 2012–2022), the effort only targets free‐swimming, age‐2 and older Lake Trout. We developed a complementary suppression method using organic (soy and wheat) pellets to cause Lake Trout embryo mortality and reduce recruitment from spawning areas. The entire Carrington Island spawning reef (0.5 ha) was aerially treated with 3.56 and 3.00 kg/m2 of pellets in 2019 and 2020, respectively. Pellet decomposition caused dissolved oxygen concentrations to decline to lethal levels at 20 cm depth in the substrate, and pellets mostly dissipated from the reef within 12 d. Lake Trout fry trap CPUE was reduced to zero after ice‐off each spring after the treatments. Prior to the treatments, 71 fry were captured during 58 trap‐nights of effort in 2017–2019. After the treatments, no fry were captured during 273 trap‐nights in 2020 and 2021. Lake Trout CPUE in large‐mesh gill nets set near Carrington Island in September did not decline during 2017–2021 and fry were again trapped on the reef in spring 2022, suggesting that adults were not deterred from spawning there in the years after the pellet treatments. Complementary methods that increase mortality of prerecruits may allow for a reduction in gill‐netting effort and the long‐term costs of maintaining Lake Trout population suppression in Yellowstone Lake. Treatment of spawning areas may improve suppression efficiency for Lake Trout and invasive fish populations elsewhere because entire cohorts are targeted while immobile and temporarily concentrated in relatively small areas.

Cocktail effects of emerging contaminants on zebrafish: Nanoplastics and the pharmaceutical diphenhydramine

Nanoplastics (NPLs) became ubiquitous in the environment, from the air we breathe to the food we eat. One of the main concerns about the NPLs risks is their role as carrier of other environmental contaminants, potentially increasing their uptake, bioaccumulation and toxicity to the organisms. Therefore, the main aim of this study was to understand how the presence of polystyrene NPLs (∅ 44 nm) will influence the toxicity (synergism, additivity or antagonism) of the antihistamine diphenhydramine (DPH), towards zebrafish (Danio rerio) embryos, when in dual mixtures. After 96 hours (h) exposure, at the organismal level, NPLs (0.015 or 1.5 mg/L) + DPH (10 mg/L) induced embryo mortality (90%) and malformations (100%) and decreased hatching (80%) and heartbeat rates (60%). After 120 h exposure, NPLs (0.015 or 1.5 mg/L) + DPH (0.01 mg/L) decreased larvae swimming distance (30–40%). At the biochemical level, increased glutathione S-transferases (55–122%) and cholinesterase (182–343%) activities were found after 96 h exposure to NPLs (0.015 or 1.5 mg/L) + DPH (0.01 mg/L). However, catalase (CAT) activity remained similar to the control group in the mixtures, inhibiting the effects detected after the exposure to 1.5 mg/L NPLs alone (increased 230% of CAT activity). In general, the effects of dual combination – NPLs + DPH (even at concentrations as low as 10 μg/L of DPH) – were more harmful than the correspondent individual exposures, showing the synergistic interactions of the dual mixture and answering to the main question of this work. The obtained results, namely the altered toxicity patterns of NPLs + DPH compared with the individual exposures, show the importance of an environmental risk assessment considering NPLs as a co-contaminant due to the potential NPLs role as vector for other contaminants.

Bacterial chondronecrosis with osteomyelitis related Enterococcus cecorum isolates are genetically distinct from the commensal population and are more virulent in an embryo mortality model

Bacterial chondronecrosis with osteomyelitis (BCO) is a common cause of broiler lameness. Bacteria that are found in BCO lesions are intestinal bacteria that are proposed to have translocated through the intestinal epithelium and have spread systemically. One of the specific bacterial species frequently isolated in BCO cases is Enterococcus cecorum. In the current study, caecal isolates were obtained from birds derived from healthy flocks (12 isolates from 6 flocks), while isolates derived from caeca, colon, pericardium, caudal thoracic vertebrae, coxo-femoral joint, knee joint and intertarsal joint (hock) were obtained from broilers derived from BCO outbreaks (111 isolates from 10 flocks). Pulsed field gel electrophoresis was performed to determine similarity. Clonal E. cecorum populations were isolated from different bones/joints and pericardium from animals within the same flock, with intestinal strains carrying the same pulsotype, pointing to the intestinal origin of the systemically present bacteria. Isolates from the intestinal tract of birds from healthy flocks clustered away from the BCO strains. Isolates from the gut, bones/joints and pericardium of affected animals contained a set of genes that were absent in isolates from the gut of healthy animals, such as genes encoding for enterococcal polysaccharide antigens (epa genes), cell wall structural components and nutrient transporters. Isolates derived from the affected birds induced a significant higher mortality in the embryo mortality model as compared to the isolates from the gut of healthy birds, pointing to an increased virulence.

Forecast-based action for conservation.

Extreme weather events pose an immediate threat to biodiversity, but existing conservation strategies have limitations. Advances in meteorological forecasting and innovation in the humanitarian sector provide a possible solution-forecast-based action (FbA). The growth of ecological forecasting demonstrates the huge potential to anticipate conservation outcomes, but a lack of operational examples suggests a new approach is needed to translate forecasts into action. FbA provides such a framework, formalizing the use of meteorological forecasts to anticipate and mitigate the impacts of extreme weather. Based on experience from the humanitarian sector, I suggest how FbA could work in conservation, demonstrating key concepts using the theoretical example of heatwave impacts on sea turtle embryo mortality, and address likely challenges in realizing FbA for conservation, including establishing a financing mechanism, allocating funds to actions, and decision-making under uncertainty. FbA will demand changes in conservation research, practice, and governance. Researchers must increase efforts to understand the impacts of extreme weather at more immediate and actionable timescales and should coproduce forecasts of such impacts with practitioners. International conservation funders should establish systems to fund anticipatory actions based on uncertain forecasts.

Embryotoxicity and Teratogenicity of Steroidal Saponin Isolated from Ophiopholis mirabilis.

Benthic invertebrates produce secondary metabolites that serve as defenses against consumers and promote their fitness. To explore the chemical defense in marine benthic echinoderms, the chemical constituents of Ophiopholis mirabilis were investigated. A steroidal monoglycoside, asterosaponin P1, was isolated from O. mirabilis for the first-time using column chromatography. The chemical structure was characterized by spectroscopic and spectrometric methods. The embryotoxicity and teratogenicity of the isolated compound were assessed using the zebrafish embryo assay, a powerful vertebrate animal model system to study mechanisms of toxicity. When applied at high concentrations, asterosaponin P1 causes a significant increase in embryo mortality. A moderate LC50 of asterosaponin P1 appeared to be time- and concentration-dependent in its toxicity to zebrafish embryos. Teratogenicity in zebrafish embryos also included morphological defects, decreased hatchability, and a reduced heart rate. These findings revealed that steroidal saponin extracted from O. mirabilis exhibited acute toxic effects on zebrafish embryos, suggesting a potential chemical defense function in marine habitats.

Daya Tetas Telur Hasil Persilangan Ayam Bangkok dan Ayam Ras Petelur (Isa brown) Menggunakan Ekstrak Daun Sirih Sebagai Bahan Sanitasi

Abstrak. Using betel leaf extract as a sanitizer, this study aims to examine the hatchability of eggs resulting from crosses between Bangkok chickens and laying hens. This study used a randomized design with four treatments and five replications. There are five eggs in each repetition. The treatments tried were soaking eggs in 70% ethanol (P0), 5% betel leaf extract (P1), 10% betel leaf extract (P2), and 15% betel leaf extract separately (P3). in this study were organisms that had not yet developed practicality, mortality and hatchability. Analysis of variance using to tabulate and analyzed the data obtained. The Least Significant Difference Test using to continue the evaluation if the treatment has a significant effect on the variables studied. The mean embryo survival rates were 88% (P0), 91% (P1), 84% (P2), and 58% (P3), according to the findings. In each treatment, the average mortality rate was 12% (P0), 9% (P1), 16% (P2), and 42% (P3). hatchability was 56% (P0), 79.% (P1), 63% (P2), and 42% (P3). The use of betel leaf extract with different concentrations as egg cleaner from crosses between Bangkok and laying hens had a significant impact on embryo survival and mortality, but not on hatchability. The use of 5% betel leaf extract as a cleansing agent for hatching eggs showed that the embryos lived longer and died less frequently.