Immunohistochemical and qPCR determination of the expression and serum level of anti‐Müllerian hormone in pre‐pubertal,intact and ovarian remnant syndrome detected bitches

The aim of this study was to determine the serum concentrations, ovarian presence and expression of anti‐Müllerian hormone (AMH) in pre‐pubertal, bitches with signs of ovarian remnant syndrome (ORS) and intact bitches. In addition, we aimed to verify the suitability of serum AMH concentrations for diagnostic purposes in sterilized bitches and/or in suspected cases of ORS in the field of veterinary medicine. For this purpose, 36 healthy female dogs divided into six groups: proestrus, oestrus, dioestrus, anoestrus, pre‐pubertal and ORS. Serum AMH concentrations were determined by electrochemiluminescence immunoassay, and ovarian presence and distribution of AMH was confirmed by immunohistochemical and qPCR techniques. According to the results of qPCR, while the expression values of AMH were at the highest concentrations in the proestrus and oestrus, there was a statistically significant decrease in these values at the later stages of the cycle (p < 0.05). According to hormone analysis, the serum AMH values of the ORS group had decreased significantly compared with the proestrus and oestrus (p < 0.05). Although serum AMH levels of ORS group were increased compared with anestrus and pre‐pubertal groups, this increase was statistically non‐significant (p > 0.05). Immunohistochemically, AMH expression was first observed in the granulosa cells of primordial follicles in folliculogenesis. Expression values were the highest in the proestrous and oestrus groups, but values from bitches in later stages of the cycle were statistically significant decrease in comparison with these groups (p < 0.05). As a result, AMH concentration and expression were found to be higher in proestrus and oestrus than in other periods (p < 0.05). In addition, the measurable level of AMH concentration in bitches with ORS is an indication that it can be used in the diagnosis of ORS.     

Phytochemical accumulation and antioxidant capacity at four maturity stages of cranberry fruit

Specific components of cranberry fruit are being associated with human health attributes, such as maintenance of urinary tract health and antioxidant status. Some of the chemical properties and antioxidant capacity of cranberry (Vaccinium macrocarpon Ait. cv. Pilgrim) fruits were investigated at light green, blush, light red and dark red maturation stages. Fruit total phenolics, total monomeric anthocyanins, soluble solids, titratable acidity and individual organic acids were examined. Antioxidant capacity of fruits were determined by both the ferric reducing antioxidant power (FRAP) and trolox equivalent antioxidant capacity (TEAC) assays. The fruit color was measured using a portable chromameter. A converse relationship was found between total phenolics and anthocyanin as fruits mature. Total phenolic concentration was declined from 7990 to 4745 mg GAE/kg fw, while total monomeric anthocyanin content was increased from 0.8 to 111.0 mg/kg fw from green to dark red stage. Brix was increased from 6 to 9.3% as well. The main organic acid was citric acid determined by the HPLC method. The antioxidant capacity of cranberries decreased to light red stage; when a fruit accumulates more anthocyanin the activity increased again in both FRAP and TEAC methods. Averaged antioxidant capacity measured was 12.61 and 17.48 mmol TE/kg fw by FRAP and TEAC methods.     

Use of Canopy Temperature Measurements as a Screening Tool for Drought Tolerance in Spring Wheat

Remotely sensed infrared canopy temperatures provide an efficient method for rapid, non-destructive monitoring of whole-plant response to water stress. Field studies were conducted in 1992 and 1993 to evaluate the potential for using canopy temperatures to screen for drought tolerance in wheat. In both years, 12 spring wheat (Triticum aestivum L.) genotypes were grown under two irrigation levels (well-watered and moisture-stressed) imposed between tillering and anthesis with a line-source sprinkler irrigation system. Canopy temperature measurements were taken in well-watered and moisture-stressed plots between 1330 and 1430 h (MDT) on five clear days in 1992 and seven clear days in 1993 during the late vegetative and early reproductive growth periods following closure of the plant canopy. Genotypes exhibited differences in mean canopy temperatures across the two irrigation levels and two years. Klasic consistently had the highest canopy temperature under moisture-stressed conditions, while Bannock and Pondera had the lowest. Bannock, Yecora Rojo and Klasic had the warmest canopies under well-watered conditions, while Vandal, Amidon and Rick had the coolest. Plot-to-plot variation in canopy temperature under water stress conditions was evident for differences in grain yield. Significant correlations between canopy temperature and yield under moisture-stress conditions and drought susceptibility index values indicated the potential for screening wheat genotypes for drought response.     

Effect of Dietary Supplementation of Acidic Calcium Sulfate (Vitoxal) on Growth,Survival, Immune Response and Gut Microbiota of the Pacific White Shrimp,Litopenaeus vannamei

Dietary acidifiers have been recognized as beneficial in animal production including aquacultural production of fish where they confer such benefits as improved feed utilization, growth, and resistance to bacterial pathogens. If improvements in growth and immune responses by acidifier supplementation can be confirmed in shrimp, then mortalities due to diseases could be minimized, limiting the emergence of disease‐resistant bacterial pathogens as a potential result of antibiotic misuse. With this in mind, a 35‐d feeding trial was conducted to evaluate growth, enteric microbiota populations, and nonspecific immune responses of Litopenaeus vannamei fed diets containing the commercial acidifier Vitoxal, based on acidic calcium sulfate in an indoor temperature‐controlled, recirculating culture system without any natural productivity. Experimental diets were formulated to contain 0 (basal), 0.4, 1.2, 1.6, or 2.0% acidic calcium sulfate (ACS) by weight. Shrimp fed in excess, 15 times a day using automatic feeders. Weight gain and survival among treatments were excellent, but not significantly different (P > 0.05). Denaturing gradient gel electrophoresis analysis revealed that the enteric microbial community of shrimp fed the basal diet differed markedly from those fed the acidifier on the basis of 64.9% similarity coefficient. Shrimp fed the commercial acidifier at 1.2 and 2.0% responded significantly (P < 0.05) better to reduced stress and displayed enhanced immune responses including hemocyte phagocytic capacity, hemolymph protein concentration, hyaline cell counts, and hemolymph glucose, compared with shrimp fed the basal diet. These results point to an enhanced performance in terms of positive shifts in the composition of enteric microbial communities as well as improved immune performance, with no changes in growth or survival.     

Factors affecting the reproductive efficiency in crayfish: implications for aquaculture

In recent years, crayfish aquaculture industry has been developing rapidly in different regions of the world. Crayfish aquaculture is often carried out in dams, lakes and extensive pond systems. There is an increasing demand for crayfish juvenile in aquaculture sector. Therefore, it is important to know which factors affect reproductive efficiency in crayfish. In the present review, factors affecting the reproductive efficiency of crayfish are divided into two main sections, external and internal factors. Dietary lipids, phospholipids, highly unsaturated fatty acids, protein and amino acids, vitamins and carotenoids are important external factors affecting broodstock crayfish reproduction. In addition, various external (i.e. female size, stocking density, temperature, photoperiod, sex ratio) and internal (i.e. endogenous hormones) factors affect the reproductive efficiency in crayfish. This study reviews the present knowledge with the purpose of realizing the factors that are important to reach optimal crayfish nutrition, maturation and reproduction (i.e. pleopodal egg number and quality). This review will particularly be useful for crayfish farmers and crayfish hatchery units.     

Determination of selected antibiotics in farmed fish species using LC‐MS/MS

The aim of the study was to monitor the occurrence of 37 antibiotics from different classes including fluoroquinolone, tetracycline, macrolide, sulphonamide, penicillin, amphenicol and cephalosporin in three different farmed fish species (n = 75; rainbow trout, see bass, gilthead see bream) from five cities in Turkey. A method for LC‐MS/MS was optimized and validated to assess these antibiotics. As a result, 25 samples showed contamination with at least one antibiotic residue. The most detected antibiotics were norfloxacin (NOR) (18.7%), marbofloxacin (MAR) (12%), tetracycline (TC) (10.7), danofloxacin (DAN) (9.3%), oxytetracycline (OTC) (9.3%), tilmicosin (TLM) (8%) and enrofloxacin (ENR) (6.7%). A total of 17.3% of samples examined were found to be contaminated with residues exceeding the maximum residue limits (MRLs). The results indicate that antibiotics are frequently used in aquaculture production without the lack of knowledge. This study also revealed that the consumption of farmed fish could produce a public health concern as the level of residues for some antibiotics was much higher than MRLs.     

Changes in the digestive enzymes and hormones of gilthead seabream larvae (Sparus aurata, L. 1758) fed on Artemia nauplii enriched with free lysine

Variations in digestive enzymes and hormones during the larval development of gilthead seabream (Sparus aurata) fed on live prey (Artemia nauplii) enriched with free lysine were investigated for 16 days (from day 24 to day 40). Prior to initiation of the experiment, newly hatched larvae were transferred from incubators to fiber glass tanks (300 l) with black walls and fed the same diet until day 24. Each experiment was performed in triplicate. The amount of free lysine in the Artemia nauplii was increased by adding a 5.3 mM free amino acid solution to the culture water during a 16-h enrichment period. Larvae were sampled four times at four-day intervals, for 16 days. Larvae fed on the control group had lower growth than those fed on the lysine group at the end of the study but the difference was not significant (P > 0.05). The highest trypsin activity and lap/leu–ala peptidase ratios were observed in the control group (P < 0.05). Significant differences between bombesin activities of the treatment groups were not found in the fifth minute after the start of feeding (P > 0.05) but were significant in the fifteenth minute (P < 0.05). Significant differences between the CCK levels of the treatment groups were found (P < 0.05). In conclusion, three main ideas are presented in this study. First, Artemia nauplii can successfully be enriched with free lysine. Second, bombesin (GRP) and cholecystokinin (CCK) activity can be stimulated by free lysine. Third, the mechanisms controlling adaptation of the activity of the trypsin to the amount of dietary protein were not activated until day 40.     

The changes in digestive enzymes and hormones of gilthead seabream larvae (Sparus aurata, L 1758) fed on Artemia nauplii enriched with free methionine

Variations in digestive enzymes and hormones during the larval development of gilthead seabream (Sparus aurata) fed on live prey (Artemia nauplii) enriched with free methionine were investigated for 16 days (from day 24 to day 40). Prior to initiation of the experiment, newly hatched larvae were transferred from incubators to fiber glass tanks (300 l) with black walls and fed with same diets until day 24. Each experiment was performed in triplicate. In the experimental group, the content of the free methionine in the Artemia nauplii was increased by adding a 5.3 mM free methionine solution to the culture water during a 16-h enrichment period. The larvae of both the control and enriched-methionine groups were sampled four times, with 4-day intervals between samplings, during a 16-day period. The larvae in the control group had a significantly lower growth than those of the methionine group at the end of the study (P < 0.05). The highest trypsin activity and leucine aminopeptidase N/leucine–alanine peptidase ratios were observed in the control group. A significant difference between bombesin activities in the treatment groups was not found at 5th minute after the initiation of feeding (P > 0.05), but they were significant at 15th minute post-initiation of feeding (P < 0.05). A significant difference between the cholecystokinin levels of the treatment groups was found (P < 0.05).     

A comparison of the growth and survival of two freshwater crayfish species, Astacus leptodactylus Eschscholtz and Pacifastacus leniusculus (Dana), under different temperature and density regimes

Growth experiments carried out with two juvenile crayfish species, Astacus leptodactylus Eschscholtz and Pacifastacus leniusculus (Dana), at different temperatures and densities highlighted the problem of cannibalism under conditions aimed at intensifying crayfish production. Cannibalism proved to be much lower in A. leptodactylus than P. leniusculus, suggesting that the former might be the better candidate for astaciculture. In the first of two population survival trials, A. leptodactylus showed a higher survival rate than P. leniusculus, with 26% of P. leniusculus surviving in the first replicate, compared to 42.3% of P. leniusculus, and 40.76% surviving in the second replicate, compared to 47.6% of P. leniusculus. The same trend was observed in the second experiment, with P. leniusculus showing survival rates of 48.5 and 55.3% (first and second replicates, respectively) in comparison to 65.2 and 68.2%, respectively, for A. leptodactylus. It is therefore highly likely that if P. leniusculus were to become established in Turkey, it would outcompete the native A. leptodactylus even if it were not to be devastated by the crayfish plague that has decimated the native species. The experiments also highlighted the problem of differential growth, with some juveniles hardly growing at all while others reached a relatively large size. The range in carapace length (CL) was 9–18 mm for A. leptodactylus and 8.5–18.5 mm CL for P. leniusculus at the end of the first experiment. While both species grew quickly, P. leniusculus hatched earlier, giving it an advantage over A. leptodactylus; consequently, by the end of the summer, the juveniles of the former were larger than those of A. leptodactylus. Specific growth rate values showed that the juveniles of the two species had similar growth rates. The results also revealed that growth was not significantly affected by density in both species at 15°C, but at 25°C, growth was significantly better at a density of 234 juveniles m⁻² than at 468 juveniles m⁻², and better at 468 juveniles m⁻² than at 937 juveniles m⁻² (P < 0.01 and P < 0.001, respectively). We conclude that P. leniusculus is a good candidate for aquaculture as it has a rapid growth rate and early hatching and maturity; however, its aggressive behavior may make it a less attractive proposition than A. leptodactylus, which is also fast growing but less aggressive. This study is a part of PhD study of M.M. Harlıoğlu, who is supported by Fırat University Elazığ, Turkey.