Effects of Karenia brevis diet on RNA:DNA ratios and egg production of Acartia tonsa

Karenia brevis is a harmful alga associated with deleterious effects on zooplankton, but the exact cause (e.g. toxin, nutritional inadequacy or starvation) of these adverse effects is not clear. RNA:DNA ratios, fecundity and fecal pellet production of Acartia tonsa were measured on mono-algal and mixed-algal culture diets of K. brevis and Peridinium foliaceum to examine the usefulness of RNA:DNA ratios as an indicator of nutrition and to determine if adverse effects of K. brevis are due to the presence of toxins, poor nutritional quality or starvation. RNA:DNA ratios and egg production values were significantly higher for 100% P. foliaceum diet compared to 100% K. brevis diet. Significant differences in egg production, but not RNA:DNA ratios, were found between the various mixed diets, suggesting egg production is a more sensitive indicator of nutritional quality than RNA:DNA ratios. Changes in RNA:DNA ratios, fecundity and fecal pellet production of copepods fed two different toxic K. brevis strains were nearly identical, indicating that the presence of brevetoxins has little affect on A. tonsa. The similarity in RNA:DNA ratios, egg production, percent hatching and fecal production between the 100% K. brevis diet and starved copepods suggests that A. tonsa does not consume K. brevis when offered as its sole food source.     

Huffmanela selachii n. sp. (Nematoda: Trichosomoididae: Huffmanelinae): A new species infecting the skin of the great hammerhead shark (Sphyrna mokarran) and the blacktip reef shark (Carcharhinus melanopterus) in the Arabian Gulf,off-shore Saudi Arabia

From January 2017 - December 2019, 75 out of 850 (8.8 %) great hammerhead sharks from the Arabian Gulf had skin lesions of black irregular discolorations on the ventral surface of the head. The lesions consisted of pencil-like lineations often advancing forward by about 2 mm in back-and-forth looped scribbles often forming a relatively linear bands of about 5–7 cm wide. Similar lesions were also found in the blacktip reef shark from the same area within the same period, and consisted of straight to irregular black lines, extended indiscriminately across the skin of the sharks. Microscopic examination of the skin revealed the presence of dark-brown eggs exhibiting the spindle or ellipsoidal eggs characteristic of Huffmanela sp. The morphometrics of eggs from both hosts were similar (62.9–89.9 μm long and 29.3–56.1 μm wide). The eggshells were smooth with polar plugs protruding or not, with an abruptly truncated crown-like or shoulder-like collar surrounding the plug. The eggs were only found in the epidermal layer of the skin. Based on the unique morphometrics of the eggs, we report a new species, named: Huffmanela selachii n. sp.. This appears to be the first report of Huffmanela from either the great hammerhead shark or the blacktip reef shark, and the third reported Huffmanela in sharks from the Arabian Gulf. It is also one of few species reported from connecting waters of the greater Indian Ocean. This new finding contributes to our understanding of the diversity and ubiquity of Huffmanela sp. in marine creatures.     

Effect of weight and storage time of broiler breeders’ eggs on morphology and biochemical features of eggs,embryogenesis, hatchability,and chick quality

The transfer of hatchability results obtained under experimental conditions to the commercial ground with a positive financial effect proves the value and usefulness of these data. On the other hand, finding results on commercial processes of broiler breeders’ egg incubation in the literature is challenging. The presented study aimed to determine the effects of egg weight and storage time on the physical, biochemical characteristics of hatching eggs, embryogenesis and hatchability in Ross 308 broiler breeders. On the laying day, the eggs were divided into four weight groups: S – small eggs (57–61 g), M – medium eggs (62–66 g), L – large eggs (67–71 g), and XL – extra-large eggs (72–76 g). The eggs were then stored for 3, 7, 14, and 21 days under controlled conditions. As the egg storage time increased, a decrease in the yolk quality (lower index) was observed. The highest Haugh units were found in eggs from the S and M groups. The cholesterol content of the M, L, and XL groups was lower on days 7, 14, and 21 as compared to that of eggs only stored for 3 days. Egg weight loss during incubation decreased with an increase in the egg weight. An extension of the egg storage time caused an increase in the loss of egg weight. On the 14th and 18th days of hatching, an increase in the eggshell temperature was noted with an increase in the weight of the egg. The eggs stored for 7 days were characterised by the highest shell temperature on each day. The highest hatchability percentage was recorded for the M group. The hatchability rate decreased with the prolongation of the storage time, while the number of crippled chicks after hatching increased. The results confirmed that the increased weight of the eggs and prolonged storage time (14 and 21 days) increased the weight and decreased the length of the newly hatched chicks, respectively. Chicks from the heaviest eggs and those stored for 14 and 21 days showed poor results on the Pasgar score® test. The observations indicate the need to adopt various (of those available) methods to assess the quality of newly hatched chicks in hatcheries in order to produce high-quality broiler chickens. The results also indicate that prolonged egg storing beyond 14 days may affect the thyroid hormone economy during the hatching of chicks, especially in the XL group.     

Untersuchungen über den Vitamin B2-Bedarf der Legehenne

Because of the well established function of carnitine possible effects of carnitine were studied in poultry. In trial I it was investigated if carnitine and its precursors (lysine, methionine) reduce the formation of abdominal fat in broilers. Chickens (10 groups of 10 chickens each) were fed different diets (control, lysine and methionine in excess and deficient, respectively, with or without 5% fat supplement, L‐carnitine and DL‐carnitine supplement, respectively).Performance (body weight gain, feed conversion), amount of abdominal fat and carnitine concentration in blood, muscles (M. sartorius, M.pectoralis superficialis, cardiac), liver and kidney were determined. Performance and abdominal fat were influenced by dietary fat, lysine and methionine as expected and were not altered by carnitine. Excess and deficiency of lysine and methionine did not influence, fat supplement reduced and carnitine supplementation significantly increased tissue concentration of carnitine.

In trial II it was studied if supplementation of a commercial layers’ ration with either 500 mg L‐carnitine or 500 mg nicotinic acid or both per kg reduces the cholesterol concentration in yolk. Influence on body weight, feed intake, laying performance, serum and yolk cholesterol concentration could not be observed, but yolk concentration of carnitine was significantly increased in supplemented groups.

Trial III should clarify if the L‐carnitine content in broiler parentstock ration influences hatchability. Four groups of 1350 hens each were fed a commercial all‐mash supplemented with 0, 20, 50 and 100 mg L‐carnitine, respectively. Hatching rate was increased from 83% to 87% and from 82.4% to 85.3% in groups supplemented with 50 and 100 mg L‐carnitine, respectively, and in randomly sampled eggs of these groups carnitine concentration in yolk was higher.     

Effect of thermal stress on fertility and egg quality of Japanese quail

Heat stress is one of the major causes of a decreased performance of laying quail in tropical and subtropical countries. The aim of this study was to investigate the impact of temperature humidity index (THI) on fertility aspects, external and internal egg quality parameters in Japanese quail. One hundred and forty four (144) Japanese quail, 12 of weeks age, were used. Birds were divided randomly into three equal groups, control (at low THI, lower than 70), H₁ (at moderate THI, 70–75) and H₂ (at high THI, 76–80). Quail in the control and H₁ groups had significant greater fertility (p=0.021) and hatchability % (p=0.037), compared with H₂ group. Quail in the control group (at low THI) laid heavier egg weight with a higher external (egg weight (p=0.03), shell thickness, shell weight, eggshell ratio and eggshell density (p=0.001)) and internal egg quality score (albumin weight (p=0.026), yolk height (p=0.003), yolk index (p=0.039) and Haugh unit (p=0.001)). Otherwise, such quality traits were compromised in heat-stressed quail. At the high THI level, egg weight had a significant positive correlation with albumin weight (r=0.58, p<0.01), yolk weight (r=0.22, p<0.05), albumen ratio (r=0.17, p<0.05), yolk height (r=0.14, p<0.05) and yolk index (r=0.18, p<0.05), but was negatively correlated with yolk ratio (r=−0.15, p<0.05). Japanese quail exposed to heat stress (THI over 75) revealed drop in fertility indices and egg quality traits, indicating a detrimental policy of economic income.     

Reproductive activity and physiological status of the calanoid copepods Calanus helgolandicus and Calanoides carinatus under food-limiting conditions

The reproductive activity and the physiological state of the calanoid copepods Calanus helgolandicus and Calanoides carinatus were investigated off the coast of NW Spain during autumn to evaluate the effect of short food resources on both populations. Phytoplankton biomass was low, and neither phytoplankton size distribution nor composition was suitable to support high reproductive rates. Accordingly, egg production rates (EPR) were much lower than maximum rates for both species, pointing to food limitation. The reproductive index (RI), which represents the proportion of females with mature gonads, was < 50% at each of the three zones into which the sampling area was divided (coast, shelf and ocean). Potential recruitment rates were very low except at some nearshore stations, where the highest concentrations of chlorophyll-a (Chl-a), diatoms, dinoflagellates and large cells were found. EPR of C. helgolandicus and C. carinatus were correlated with phytoplankton biomass and unaffected by temperature. Phytoplankton carbon ingestion explained ca. 50% of the variability in EPR for both species. At most of the stations, herbivory was insufficient to cover the carbon requirements for reproduction and respiration, so females probably fed on heterotrophic prey to meet their demands. However, given the low fecundity observed, this omnivorous diet did not seem to be optimum for reproduction, and a severe food limitation is thus suggested. Furthermore, the high C/N values measured point to a notable lipid storage, but given the low EPR found, lipid reserves were probably invested into female maintenance rather than into gonad maturation. C. helgolandicus and C. carinatus populations did not mirror phytoplankton biomass distribution, but they correlated well when considering only copepodites V (CV). The CV could be preparing for the overwintering, storing lipid reserves to ensure a successful diapause, and they could also be advected by the poleward current detected during the study. Females showed a diel feeding rhythm, with highest ingestion rates during night. From our results, it follows that C. helgolandicus and C. carinatus females did not perform diel vertical migrations. We suggest that this behaviour is likely due to the food-limiting conditions, which make it more advantageous to remain at the surface during daytime.     

Lysine-derived cross-links in the egg shell membrane

Egg shell membrane protein contains significant quantities of the lysine-derived aldehyde, allysine, and its aldol condensation product. NaB³H₄ reduction followed by alkaline hydrolysis of purified protein revealed that there were six residues/1000 of both allysine and the reduced aldol while only traces of desmosine and isodesmosine were detected. The amino acid composition of the membrane protein did not resemble that of mammalian elastin.     

The dual roles of Armigeres subalbatus prophenoloxidase V in parasite melanization and egg chorion melanization in the mosquito Ar. subalbatus

Phenoloxidases (POs) play key roles in various physiological functions in insects, e.g., cuticular sclerotization, wound healing, egg tanning, cuticle formation and melanotic encapsulaction of pathogens. Previously, we identified five POs, designated As-pro-PO I–V, from the mosquito Armigeres subalbatus and demonstrated that the functions of As-pro-PO I, II and III, were associated with filarial parasite melanization, blood feeding and cuticle formation, respectively. In the present study, we delineate the dual functions of As-pro-PO V. We found that the level of As-pro-PO V mRNA in mosquitoes was significantly increased after microfilaria challenge or blood feeding, and decreased to normal level after oviposition. Knockdown of As-pro-PO V by dsRNA resulted in significant decreases in the degree of microfilaria melanization, egg chronic melanization rates and egg hatching rates in Ar. subalbatus. Further transfection and electrophoretic mobility-shift assays verified the As-pro-PO V gene might regulated by both AP-1, a putative immune-related regulatory element and CdxA, a developmental regulatory element. The binding of AP-1 and CdxA motif with mosquito nuclear extracts was significantly enhanced after microfilaria challenge and blood-feeding in Ar. subalbatus, respectively. These results indicate that As-pro-PO V is a critical enzyme that is required for both an effective melanization immune response and egg chorion melanization in this mosquito.