Mohamed M.M. Kandiel

Modulation of genotoxicity and endocrine disruptive effects of malathion by dietary honeybee pollen and propolis in Nile tilapia (Oreochromis niloticus)

The present study aimed at verifying the usefulness of dietary 2.5% bee-pollen (BP) or propolis (PROP) to overcome the genotoxic and endocrine disruptive effects of malathion polluted water in Oreochromis niloticus (O. niloticus). The acute toxicity test was conducted in O. niloticus in various concentrations (0–8 ppm); mortality rate was assessed daily for 96 h. The 96 h-LC50 was 5 ppm and therefore 1/5 of the median lethal concentration (1 ppm) was used for chronic toxicity assessment. In experiment (1), fish (n = 8/group) were kept on a diet (BP/PROP or without additive (control)) and exposed daily to malathion in water at concentration of 5 ppm for 96 h “acute toxicity experiment”. Protective efficiency against the malathion was verified through chromosomal aberrations (CA), micronucleus (MN) and DNA-fragmentation assessment. Survival rate in control, BP and PROP groups was 37.5%, 50.0% and 100.0%, respectively. Fish in BP and PROP groups showed a significant (P < 0.05) reduction in the frequency of CA (57.14% and 40.66%), MN (53.13% and 40.63%) and DNA-fragmentation (53.08% and 30.00%). In experiment (2), fish (10 males and 5 females/group) were kept on a diet with/without BP for 21 days before malathion-exposure in water at concentration of 0 ppm (control) or 1 ppm (Exposed) for further 10 days “chronic toxicity experiment”. BP significantly (P < 0.05) reduced CA (86.33%), MN (82.22%) and DNA-fragmentation (93.11%), prolonged the sperm motility when exposed to 0.01 ppm of pollutant in vitro and increased the estradiol level in females comparing to control. In conclusion, BP can be used as a feed additive for fish prone to be raised in integrated fish farms or cage culture due to its potency to chemo-protect against genotoxicity and sperm-teratogenicity persuaded by malathion-exposure.

Ultrasonographic assessment of fetal growth in miniature “Shiba” goats (Capra hircus)

The aim of the present study was to monitor fetal growth in relation to gestational stage to generate formulae which could be used to estimate fetal age in goats. Eight miniature Shiba goats (Capra hircus) were examined weekly by transrectal and transabdominal ultrasound scanning during the gestation period between Day 21 and 126 days of gestation. For accurate judgment, all fetometric parameters were measured at least three times per one examination for each animal. Quantification of the growth of the fetus allowed the development of a number of predictors of fetal age. Low correlations were associated with measurement of the chest diameter (R(2)=0.869), trunk diameter (R(2)=0.8969), tibia length (R(2)=0.8662) and placentome diameter (R(2)=0.8999). Moderate correlation was assessed by calculation of the length of six successive lumbar vertebrae (R(2)=0.9296), femur length (R(2)=0.9278), heart axis length (R(2)=0.9382 and 0.9589; for the longitudinal and transverse axis, respectively), occipitonasal length (R(2)=0.9527), umbilical cord diameter (R(2)=0.9119) and orbit diameter (R(2)=0.9239). A high correlation was estimated in investigating the length of six successive thoracic vertebrae (R(2)=0.9674), braincase diameter (R(2)=0.9831) and crown rump length (R(2)=0.9848). In conclusion, the intrauterine fetal biometry estimation through ultrasound might be useful to predict the accurate gestational age in miniature goats.

Effect of reproductive status on body condition score, progesterone concentration and trace minerals in sheep and goats reared in South Sinai, Egypt

This study aimed at evaluating the effect of reproductive stage on nutritional status, and hormonal and trace mineral levels in sheep and goats reared in harsh arid conditions in South Sinai, Egypt. Egyptian local breeds of sheep (n=74) and goats (n=58) raised in South Sinai were examined by means of transabdominal ultrasonography to verify single vs. multiple feti pregnancy or non-pregnant status. Serum samples were collected for assessment of progesterone and trace minerals (selenium (Se), zinc (Zn), copper (Cu) and iron (Fe)) levels. Evaluation of the nutritional status of animal flocks showed that the mean body condition score was below the average levels, but did vary noticeably with pregnancy or between sheep and goats. Serum progesterone concentrations were significantly higher in pregnant animals with multiple fetuses than those bearing single fetus (p < 0.01), and both were higher than the respective values in non-pregnant animals (p < 0.001). Serum trace mineral levels were too low with no significant differences caused by pregnancy, number of fetuses or animal species. From these results, we could conclude that the state of pregnancy and fetal number have a significant influence on serum levels of progesterone. However, the harsh conditions in South Sinai in terms of scarce precipitation, low cultivation chances and high water salinity are the major factors influencing nutritional status or trace mineral levels in the reared small ruminants, regardless of the reproductive state or species. Keywords: Goat, pregnancy, progesterone, reproductive status, sheep, South Sinai, trace minerals

Physiological roles of inhibin in regulation of FSH secretion and follicular development during early pregnancy in goats

The aim of the current study was to clarify the physiological role of inhibin in controlling FSH secretion and follicular development during the early pregnancy in goats. Eight goats investigated sonographically on Days 19-21 (Day 0=day of mating) for pregnancy were assigned into control (n=3) and treated (n=5) groups. The ovaries of all animals were daily scanned with ultrasound for follicles 2mm or more in diameter from 1 day before to 5 days after treatment. On Day 25 postbreeding; animals received either 10 ml, of normal goat serum or antiserum against [Tyr (30)]-inhibin alpha (1-30). Jugular blood samples were collected every 6 h starting 24 h before and until 120 h after treatment. The plasma concentration of FSH increased at 6 h and remained at significantly high levels until 120 h in treated vs. control group. The plasma concentrations of estradiol showed a marked increased at 66 h, with peak levels at 120 h after treatment of antiserum. The basal concentrations of LH and the pattern of plasma concentrations of progesterone were not significantly different between the two groups. The number of medium size (3.5-5.0 mm) follicles increased considerably from Day 2, whereas small (3.5 mm or less) and large (5 mm or more) follicles increased noticeably from Day 3, as compared with pre-treatment and controls. These results clearly indicated that inhibin is a key hormone in regulation of follicular development through regulation of endogenous FSH secretion during early pregnancy in goats.

PCR-RFLP of bone morphogenetic protein 15 (BMP15/FecX) gene as a candidate for prolificacy in sheep

Abstract Bone morphogenetic protein 15 (BMP15/FecX) gene is considered one of the major genes and a candidate marker for the reproduction in farm animals, especially sheep. The present study aimed to detect the genetic polymorphisms of BMP15 gene in sheep using PCR-RFLP technique. In the present study, 115 ewes were assigned into high and low prolificacy categories according to their reproductive history. In high prolific group (n = 20), ewes produced twins more than single births. In the low prolific type (n = 95), the ewes produced single births more than twins. DNA was extracted from blood samples of all ewes, subjected to PCR-RFLP analysis and confirmed by sequence analysis. The PCR products of 356 bp size were cut with HinƒI restriction enzyme. Three digested fragments of 70, 117 and 169 bp were obtained in both types of sheep. All animals were homozygous with CC genotype. In conclusion, the accessible findings did not detect any mutation in FecX gene in sheep, regardless their prolificacy. Therefore, further attempts are necessary to detect other SNP for BMP-15 gene in Egyptian sheep breeds.

Effect of epidermal growth factor on buffalo frozen spermatozoa biometry and metabolic activity

Objective: To assess the effects of epidermal growth factor (EGF) on the Egyptian buffalo bull frozen semen, EGF was incorporated at 0 (control), 50, 100, 200 and 400 ng/mL of extender (Bioxcell®). Methods: Semen features, spermatozoa biometry, total liberated amounts of enzymes (aspartate transaminase, alanine aminotransferase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) and lipid peroxidation markers (thiobarbituric acid reactive substances, malondialdehyde, glutathione peroxidase, nitric oxide, catalase (CAT) and superoxide dismutase (SOD)) were determined in the spermatozoa-free extracellular extender. Results: Spermatozoa membrane integrity significantly (P<0.05) increased, but DNA integrity decreased with EGF 200 ng/mL. Spermatozoa head (dimensions, area and perimeter), but not shape, as well as acrosome and midpiece measures substantially differed with regard to EGF. Principle piece length and volume markedly decreased (at 100 and 200 ng/mL), while total tail/flagellum length increased (at 50 ng/mL) after EGF supplementation. EGF 50 ng/mL was associated with the decline of nitric oxide levels and catalase enzyme activity, but EGF 100 ng/mL significantly decreased the total liberated amounts of enzymes (aspartate transaminase, lactate dehydrogenase, acid phosphatase and alkaline phosphatase) as well as lipid peroxidation markers (thiobarbituric acid reactive substances and malondialdehyde). Conclusions: EGF in vitro supplementation would affect the semen characteristics of buffalo bull with 100 ng/mL counteracted the freezing mediated oxidative stress indicated with the lowest enzymes leakage and lipid peroxidation.

Histomorphometry aspect of thyroid gland and biochemical profile in pregnant and non-pregnant dromedary camels

Modulation of body physiology and metabolism is a prerequisite for successful pregnancy in camels, which is about a year and few months. The present study was carried out to verify the effect of pregnancy on histomorphometry and functional traits of thyroid gland, as well as some biochemical indices of liver and kidney functions and mineral profile in one humped camels, under Egyptian climatic conditions or desert conditions. The thyroid tissues were fixed in 10% formalin solution and processed following routine histological techniques after gross examination and biometric assessment. Serum levels of thyroid hormones (triiodothyronine, thyroxineand thyroid stimulating hormone), serum blood metabolites (total bilirubin, blood urea nitrogen and creatinine), liver enzymatic activity (aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (ALP)), and minerals (sodium, potassium, calcium and phosphorus) were estimated in pregnant (n=30) and non-pregnant (n=30) camels. There were no significant differences between the right and left thyroid lobes between pregnant and non-pregnant camel. Pregnant status in camels had insignificant influence on histological and functional traits of thyroid as well as serum blood metabolites, liver enzymes and minerals levels, as compared with non-pregnant animals during the first six months of pregnancy. Therefore, reproductive status (during early pregnancy) has no substantial effect on camel physiological mechanisms or metabolic activity. Key words: Biochemical, dromedary, histology, thyroid gland, thyroid hormones.